本發(fā)明涉及裂解鐮孢毒素的多肽變體、包含所述多肽變體的添加劑和所述多肽變體和/或所述添加劑的用途，以及涉及用于借助于所述多肽變體和/或所述包含所述多肽變體的添加劑來裂解鐮孢毒素的方法。

真菌毒素非常頻繁地出現(xiàn)在農(nóng)業(yè)植物性產(chǎn)品中，并且依據(jù)真菌毒素的種類和濃度而引起嚴(yán)重的經(jīng)濟(jì)損害，特別是在從農(nóng)業(yè)產(chǎn)品制備的食物或飼料中，和也在用這樣的食物或飼料喂養(yǎng)的動物和人中，其中這樣的損害是極其多種多樣的。已經(jīng)開發(fā)了許多方法，試圖用這些方法來使真菌毒素變得無害，也就是說使真菌毒素解毒或降解，以便在動物和人的營養(yǎng)、動物飼養(yǎng)、飼料和食物加工等的領(lǐng)域中盡可能地阻止由真菌毒素引起的損害。

突出的一組真菌毒素為鐮孢毒素(類)，其中術(shù)語“鐮孢毒素”或“鐮孢毒素類”是等價的并且各自涉及由霉菌鐮孢屬物種(fusariumsp.)所產(chǎn)生的串珠鐮孢菌素以及其衍生物和降解產(chǎn)物中的至少一種或多種或者全部，然而特別地涉及串珠鐮孢菌素a1-2(fa1-2)，串珠鐮孢菌素b1-4(fb1-4)，串珠鐮孢菌素c1、2、4(fc1、fc2、fc4)和hfc1，也涉及經(jīng)部分水解的串珠鐮孢菌素fa1-2、fb1-4、fc1-2、fc4和hfc1。經(jīng)部分水解的串珠鐮孢菌素具有僅一個丙三羧酸殘基，而fa1-2、fb1-4、fc1-2、fc4和hfc1具有兩個丙三羧酸殘基。此外，結(jié)構(gòu)上相似的番茄鏈格孢(alternariaalternatalycopersici)(aal)毒素也被鐮孢毒素組所涵蓋，其中aal毒素被再分為aal-ta1(casno79367-52-5)、aal-ta2(casno79367-51-4)、aal-tb1(casno176590-32-2)和aal-tb2(casno176705-51-4)這些組。fa1-2、fb1-4、fc1-2、fc4和hfc1具有下面的結(jié)構(gòu)式：

fb1是最經(jīng)常出現(xiàn)的來自鐮孢毒素組的毒素，然而已知許多也在人和動物中顯示出毒性效應(yīng)的衍生物和相關(guān)分子。將在人或動物中由于攝入真菌毒素而引起的疾病稱為真菌毒素中毒，在鐮孢毒素的情況下也稱為鐮孢毒素真菌毒素中毒。因而，已知鐮孢毒素通過與神經(jīng)酰胺合酶的相互作用而阻礙鞘脂代謝。鞘脂不僅是細(xì)胞膜的組成部分，而且也在許多基本的細(xì)胞過程(例如細(xì)胞生長、細(xì)胞遷移和細(xì)胞連接)中，在炎癥過程或細(xì)胞內(nèi)轉(zhuǎn)運(yùn)程序中，作為信號分子或信使分子發(fā)揮重要的作用。由于阻礙鞘脂代謝，鐮孢毒素被認(rèn)為對于在最不同的動物種類和人上的毒性效應(yīng)負(fù)有責(zé)任。因而，可以證明，鐮孢毒素以免疫抑制方式在嚙齒動物中產(chǎn)生致癌效應(yīng)，并且通過流行病學(xué)數(shù)據(jù)將它們與人中的食管癌和神經(jīng)管缺陷相聯(lián)系。在各種不同的動物種類例如豬中，它們被認(rèn)為對于典型的由中毒引起的肺水腫負(fù)有責(zé)任。關(guān)于鐮孢毒素真菌毒素中毒的例子為神經(jīng)毒性疾病，例如馬的腦白質(zhì)軟化或者在豬中的由于串珠鐮孢菌素中毒而引起的肺水腫。由于在各種不同的谷類上和特別地在玉米、堅果和蔬菜上鐮孢毒素的污染幾乎無所不在，因此它們的涉及人和動物的健康的強(qiáng)烈負(fù)面的效應(yīng)是不可忽略的。

在ep-a1860954中已經(jīng)描述了串珠鐮孢菌素的微生物降解，根據(jù)該文獻(xiàn)使用微生物來使串珠鐮孢菌素和串珠鐮孢菌素衍生物解毒，在其中向飼料中添加從精確定義的用于使串珠鐮孢菌素解毒的菌株中選擇的解毒性細(xì)菌或酵母。

也已經(jīng)描述了用于串珠鐮孢菌素的生物降解的分解代謝途徑和對此負(fù)責(zé)的基因和酶。因而，例如ep-a0988383描述了使串珠鐮孢菌素解毒的組合物和方法，其中首先在轉(zhuǎn)基因植物中產(chǎn)生所使用的降解串珠鐮孢菌素的酶，在其中串珠鐮孢菌素的解毒借助于氨基氧化酶來實(shí)現(xiàn)，所述氨基氧化酶對于其酶促活性來說需要分子氧。

此外，wo2004/085624描述了氨基轉(zhuǎn)移酶、脫氨酶和氨基變位酶以及用于以酶促方式使胺化毒素(例如串珠鐮孢菌素)解毒的方法。在此使用具有脫氨酶活性的多肽用于進(jìn)行解毒。

上面所述的產(chǎn)品或方法具有缺點(diǎn)，即對于真菌毒素的解毒來說需要分子氧以及任選地輔因子，其中特別地上述氨基氧化酶在無氧的反應(yīng)條件下不顯示出效應(yīng)。

ep-a2326713涉及添加劑，以及涉及用于制備所述添加劑的方法，用所述添加劑可成功實(shí)現(xiàn)在不依賴于氧且無輔因子的酶促反應(yīng)中降解串珠鐮孢菌素。然而，在其中所描述的和對于解毒來說決定性的酶即羧酸酯酶的溫度穩(wěn)定性是低的，以至于添加劑或seqidno.46的羧酸酯酶不適合于在高的溫度下進(jìn)行使用。

在食品工業(yè)和飼料工業(yè)中，用于制備經(jīng)衛(wèi)生化的產(chǎn)品的熱處理(在其中降低了微生物負(fù)荷)具有重大意義。在此，特別地，出于眾多原因(例如為了改善流動性，為了減少粉塵產(chǎn)生，為了降低微生物負(fù)荷，特別是沙門氏菌的微生物負(fù)荷)的飼料的?；菑V泛流行的，并且是一種標(biāo)準(zhǔn)化過程。在?；^程期間，通常借助于熱蒸汽來使物品潤濕、將其加熱并隨后在壓力下擠壓通過模具。使用多肽或酶作為待粒化的食物或飼料的添加劑是一項(xiàng)技術(shù)挑戰(zhàn)，這是因?yàn)槊富蚨嚯耐ǔτ诟叩臏囟仁敲舾械?。酶或多肽的熱處理可以?dǎo)致其比活性的下降和/或不可逆的變性。一種對此進(jìn)行抵抗的可能性是蛋白質(zhì)的包囊或包衣，例如在wo92/12645中所描述的那樣。由此可成功實(shí)現(xiàn)保護(hù)蛋白質(zhì)免于熱的影響，然而這種做法帶來這樣的風(fēng)險，即蛋白質(zhì)在口-胃-腸道中不能夠足夠快地釋放，因此其效應(yīng)要么不能夠足夠快地發(fā)揮要么完全不能夠發(fā)揮。迄今已知的用于使鐮孢毒素解毒的多肽由于其低的溫度穩(wěn)定性而不能夠在沒有事先包囊或事先包衣的情況下添加給待?；娘暳匣蚴澄?。

在其中在高的溫度下使鐮孢毒素解毒是重要的技術(shù)過程為，例如面糊制品和其他玉米產(chǎn)品(例如玉米粥、爆米花、玉米片、玉米面包或玉米粉圓餅)的生產(chǎn)，以及淀粉液化過程、糖化過程或發(fā)酵過程(例如制漿過程或發(fā)酵過程，特別是在生物乙醇制備中)。在此重要的是確保通過這些過程制備的食物或飼料不包含有害量的鐮孢毒素。迄今已知的多肽由于其在各自的工藝溫度下具有過低的活性或不存在活性而不能夠在這些過程中使用。

因此，對于用于特異性地、安全地和可靠地裂解鐮孢毒素(特別地用于使鐮孢毒素解毒)的酶或多肽存在需求，其中酶促反應(yīng)既不需要氧也不需要輔因子，并且所述酶或多肽此外還具有足夠的溫度穩(wěn)定性以及足夠的溫度活性，因此所述酶或多肽可以在處于高的溫度下的技術(shù)過程中進(jìn)行使用。

現(xiàn)在，本發(fā)明旨在提供seqidno.46的鐮孢毒素羧酸酯酶的裂解鐮孢毒素的多肽變體以供使用，用所述多肽變體可成功實(shí)現(xiàn)在不依賴于氧且無輔因子的情況下將至少一種鐮孢毒素裂解為無毒或低毒的產(chǎn)物，其中所述多肽變體具有相比于seqidno.46的鐮孢毒素羧酸酯酶而言提高的溫度穩(wěn)定性或提高的溫度活性。

為了解決該任務(wù)，本發(fā)明的特征主要在于，所述多肽變體各自具有在n-末端處縮短了47個氨基酸的氨基酸序列，其中所述氨基酸序列與seqidno.46的氨基酸序列區(qū)段48-540具有至少70％，優(yōu)選地80％，特別優(yōu)選地100％的序列同一性，即seqidno.1。已經(jīng)令人驚訝地顯示，相比于seqidno.46而言縮短了47個氨基酸的氨基酸序列不僅相比于該序列而言顯著地更具有活性，而且具有相比于該序列而言提高的溫度穩(wěn)定性。

如相應(yīng)于本發(fā)明的一個進(jìn)一步改造方案那樣，通過形成seqidno.1的多肽變體，特別是具有與seqidno.1具有至少70％的序列同一性的氨基酸序列并且在至少一個從由下列各項(xiàng)組成的組中選擇的位置處具有氨基酸置換的多肽變體：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490，獲得了相比于具有seqidno.1的鐮孢毒素羧酸酯酶而言提高了至少4％的溫度穩(wěn)定性。通過使用這樣的具有裂解鐮孢毒素的特性的多肽變體，可成功實(shí)現(xiàn)例如在處于高的溫度下的技術(shù)過程期間使鐮孢毒素解毒。這特別地通過除了溫度穩(wěn)定性外同樣也提高了的溫度活性而成為可能。特別地，由此可以確保，所述多肽變體的酶促活性在高的溫度荷載下(如例如在容器中進(jìn)行運(yùn)輸期間也可以是這種情況)也仍然保持。

名稱“羧酸酯酶”涉及所有這樣的酶、多肽或多肽變體，其能夠借助于水將羧酸酯化合物裂解為相應(yīng)的醇化合物和羧酸化合物。名稱“鐮孢毒素羧酸酯酶”涉及所有這樣的酶、多肽或多肽變體，其能夠水解至少一種鐮孢毒素，通過從中以水解方式裂解出至少一個丙三羧酸(丙烷-1,2,3-三羧酸)。在此所描述的“裂解鐮孢毒素的特性”涉及以水解方式從至少一種鐮孢毒素(特別是fa1-2、fb1-4、fc1-2、fc4、hfc1、aal-ta1-2和aal-tb1-2或者其衍生物或降解產(chǎn)物)中裂解出至少一個丙三羧酸殘基的能力。

下面所使用的術(shù)語取自專業(yè)用語并且總是以傳統(tǒng)的意義進(jìn)行使用，除非另外指出。因而，術(shù)語“多核苷酸”涉及具有所有長度和序列的每一種類型的遺傳物質(zhì)，例如單鏈和雙鏈dna和rna分子，包括調(diào)控元件、結(jié)構(gòu)元件、基因群、質(zhì)粒、全基因組及其片段。名稱“多肽”包括蛋白質(zhì)，例如酶、抗體，以及具有直至500個氨基酸的多肽，例如肽抑制劑，蛋白質(zhì)的結(jié)構(gòu)域，還有具有小的序列長度例如少于10個氨基酸的短多肽，例如受體、配體、肽激素、標(biāo)簽等。名稱“在多核苷酸或多肽中的‘位置’”涉及在所述多核苷酸或多肽的序列中單個特定的堿基或氨基酸。氨基酸的名稱縮寫為專業(yè)人員熟悉的單字母代碼或三字母代碼。

名稱“單位”或“u”涉及酶、多肽或多肽變體的催化活性的量度，并且被定義為在一分鐘內(nèi)在確定的條件下被轉(zhuǎn)化或裂解的底物(在此為串珠鐮孢菌素b1)的微摩爾數(shù)(μmol)。例如，如果在15分鐘內(nèi)從60μmolfb1中裂解出至少一個丙三羧酸殘基，那么這相應(yīng)于4個單位的催化活性。對于fb1裂解，如下來定義反應(yīng)條件：轉(zhuǎn)化反應(yīng)在具有0.1mg/ml牛血清白蛋白的20mmtris-hcl緩沖液(ph8.0)中在30℃的溫度下進(jìn)行30分鐘。在反應(yīng)中的底物濃度為100μμfb1。

將以“單位/毫升溶液”給出的酶溶液或多肽溶液或多肽變體溶液的酶濃度定義為酶溶液或多肽溶液或多肽變體溶液的“催化活性”、“催化酶活性”或“活性”。

名稱“比活性”被定義為每毫克酶、多肽或多肽變體的催化活性，并且從酶溶液的催化活性與在該溶液中酶的質(zhì)量濃度(質(zhì)量/體積單位)之比來計算出來。例如，如果酶溶液具有50u/ml的催化活性和1mg/ml的質(zhì)量濃度，那么比活性為50u/mg。

名稱“溫度穩(wěn)定性”涉及酶、多肽或多肽變體在對于高的溫度的在時間上受限制的暴露(預(yù)溫育)之后保持其催化活性的特性。通過測量在相同且確定的條件下在5分鐘的熱處理后和沒有熱處理的情況下酶溶液或多肽溶液或多肽變體溶液的活性并進(jìn)行比較來測定溫度穩(wěn)定性。因此，溫度穩(wěn)定性是關(guān)于酶對于在時間上有限制的熱作用的抵抗能力的量度。經(jīng)熱處理的酶相比于未經(jīng)熱處理的100％-對照而言的剩余活性為50％時所處的溫度為關(guān)于溫度穩(wěn)定性的量度，并且縮寫為t(50％)。例如，如果沒有預(yù)溫育的酶溶液的活性為50u/ml并且在50℃下預(yù)溫育5分鐘后為25u/ml，那么所述酶的溫度穩(wěn)定性為50℃或者說直至50℃所述酶是溫度穩(wěn)定的。多肽變體相比于seqidno.1的親本多肽而言的t(50％)的增加被定義為關(guān)于提高的溫度穩(wěn)定性的量度，并且可以相對地作為百分比值或者絕對地以攝氏度來給出。

術(shù)語“溫度活性”定義了這樣的溫度，在所述溫度下所述酶、多肽或多肽變體具有最高的活性，其中在30分鐘的時間段內(nèi)測量催化活性。

術(shù)語“多肽變體”或“變體”特別地涉及這樣的多肽序列，其相比于seqidno.46而言具有至少一個氨基酸置換，其中仍然保持了酶促功能，即裂解鐮孢毒素的特性。此外，“多肽變體”還可以相對于seqidno.46的多肽序列而言具有氨基酸插入或缺失，特別是在c-末端或n-末端處延長或縮短了的序列。如果酶促反應(yīng)機(jī)制保持不變，那么酶促功能基本上得到保持，即鐮孢毒素在相同的位點(diǎn)處被水解并且相關(guān)于seqidno.46的原始親本多肽而言所述變體的比活性為至少10％，優(yōu)選地至少50％，更優(yōu)選地至少90％，然而特別地大于>100％。

在確定位置處的氨基酸置換用下面的命名法來進(jìn)行描述：原始氨基酸；位置；新氨基酸。例如，如果在位置134處的脯氨酸被甘氨酸替代，那么這用pro134gly或p134g來指明。多重突變用加號或斜線來分開。例如，如果在位置134處的脯氨酸被甘氨酸替代和在位置136處的精氨酸被賴氨酸替代，那么這用pro134gly+arg136lys或pro134gly/arg136lys或p134g+r136k或p134g/r136k來指明。如果在一個位置處的氨基酸被兩個或更多個備選氨基酸替代，那么備選氨基酸用逗號或斜線來分開。例如，如果在位置134處的脯氨酸不僅被甘氨酸替代而且也被絲氨酸、纈氨酸和甲硫氨酸替代，那么這用pro134gly,ser,val,met或pro134gly/ser/val/met或p134g,s,v,m或p134g/s/v/m來指明。如果未詳細(xì)地定義在確定位置處的氨基酸置換或替換，那么這如此地來進(jìn)行解釋，即這些氨基酸可以被所有其他氨基酸替代。例如，如果未詳細(xì)地定義在位置134處的脯氨酸的突變，那么該脯氨酸可以被下列氨基酸之一替代：a、r、n、d、c、q、e、g、h、i、l、k、m、f、s、t、w、y或v。

名稱“序列同一性”涉及序列同一性百分比。對于氨基酸序列和核苷酸序列，序列同一性可以目視測定，但是優(yōu)選地用計算機(jī)程序來計算出來。將具有seqidno.1的氨基酸序列定義為參考序列。還在序列區(qū)段內(nèi)進(jìn)行序列比較，其中參考序列的連續(xù)序列被理解為區(qū)段。對于核苷酸序列來說，序列區(qū)段的長度通常為18至600個核苷酸，優(yōu)選地45至200個核苷酸，更優(yōu)選地100至150個核苷酸。對于肽序列來說，序列區(qū)段的長度通常為3至200個氨基酸，更優(yōu)選地15至65個氨基酸，最優(yōu)選地30至50個氨基酸。存在有許多可購得的或免費(fèi)可用的生物信息學(xué)程序，其可以被召來用于同源性測定并且持續(xù)地得到進(jìn)一步發(fā)展。對此的例子為gcgwisconsinbestfit軟件包(devereux等人,1984)、blast(altschul等人,1990)或blast2(tatusova和madden,1999)。由于這些算法的不同的設(shè)置可能性，在相同的輸入序列的情況下它們得到不同的結(jié)果是可能的。因此，必需定義搜索算法和與此有關(guān)的設(shè)置。在本情況下，序列同一性借助于程序ncbiblast(basiclocalalignmentsearchtool)來進(jìn)行測定，特別地對于多肽，用blastp，和對于多核苷酸，用blastn，其提供在“nationalcenterforbiotechnologyinformation”的主頁(ncbi；http://www.ncbi.nlm.nih.gov/)上以供使用。因此，可能的是，根據(jù)altschul等人,1997(nucleicacidsres.,25:3389-3402)的算法，將兩個或更多個序列互相進(jìn)行比較。在此，使用2014年8月12日的那個版本的程序。作為程序設(shè)置，考慮基本設(shè)置，特別地，對于氨基酸序列比較：“最大靶序列(maxtargetsequence)”＝100；“期望閾值(expectedthreshold)”＝10；“字長(wordsize)”＝3；“矩陣(matrix)”＝blosom62；“缺口成本(gapcosts)”＝“存在(existence)：11；延伸(extention)：1”；“計算調(diào)整(computationaladjustment)”＝“條件式組成得分矩陣調(diào)整(conditionalcompositionalscorematrixadjustment)”；以及對于核苷酸序列比較：字長：11；期望值(expectvalue)：10；缺口成本：存在＝5，延伸＝2；過濾器(filter)＝低復(fù)雜度激活的(lowcomplexityactivated)；匹配/錯配得分(match/mismatchscores)：2，-3；過濾器字符串(filterstring)：l；m。

根據(jù)本發(fā)明的一個進(jìn)一步改造方案，通過進(jìn)行下列氨基酸置換而由此獲得溫度穩(wěn)定性的特別大的提高：在位置10和456處的氨基酸置換選自q、e、n、h、k和r，在位置33、107、293和332處的氨基酸置換選自e、q、d、k、r和n，在位置66、463和478處的氨基酸置換選自d、e、k、n、q和r，在位置140和490處的氨基酸置換選自p、a、s和n，在位置144和367處的氨基酸置換選自i、l、m和v，在位置149、270、312、329和372處的氨基酸置換選自f、y、w和h，在位置151和453處的氨基酸置換選自d、e、k和r，在位置157和462處的氨基酸置換選自f、h、w和y，在位置199、302、365和464處的氨基酸置換選自i、l、m和v，在位置266和455處的氨基酸置換選自a、s和t，在位置267、394和429處的氨基酸置換選自a、n、p和s，在位置272處的氨基酸置換選自h、n、q和s，在位置275處的氨基酸置換選自a、d、e、g、k、n、q、r和s，在位置280處的氨基酸置換選自a、d、e、k、n、p、q、r和s，在位置284處的氨基酸置換選自a、n、p、s、t和v，在位置286處的氨基酸置換選自a、d、e、k、n、p、r和s，在位置360、377、391、419和427處的氨基酸置換選自a、i、l、s、t和v，在位置363、443和457處的氨基酸置換選自a、s、t和v，在位置364處的氨基酸置換選自h、i、l、m、n、q、s和v，在位置371處的氨基酸置換選自a、i、l、m、s、t和v，在位置389處的氨基酸置換選自i、l、m和v，在位置418、430、447和473處的氨基酸置換選自a、g和s，在位置424處的氨基酸置換選自a、d、e、g、k、r和s，在位置436處的氨基酸置換選自a、g、s和t，在位置440處的氨基酸置換選自a、g、s和t，在位置465處的氨基酸置換選自a、g、h、n、q、s和t，在位置469處的氨基酸置換選自d、e、k和r，和/或在位置487處的氨基酸置換選自n、d、q、h和s，其中在上述位點(diǎn)處原來存在的氨基酸無論如何均被替換。

根據(jù)本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案，所述氨基酸置換選自由下列各項(xiàng)組成的組：10q、33e、66d、107e、140p、144m、149f、151r、157y、199i、266s、267p、270f、272h、275e、275a、280d、280p、284t、284p、286p、286r、293e、302i、312f、329f、332e、360v、363t、364h、364l、365i、367h、371v、371m、372f、377v、389l、391v、394p、418a、419v、424a、424k、427v、429p、430a、436a、436s、440g、440s、443t、447a、453r、455s、456q、457t、462y、463d、464i、465h、465s、465g、469k、473a、478d、487n和490p。通過所述多肽變體具有至少一個這樣的置換，可以更加有針對性地提高溫度穩(wěn)定性或溫度活性，其中另外地，其他酶參數(shù)(例如比活性、ph-穩(wěn)定性或ph-活性)同樣也可以得到改善，然而其至少有著具有seqidno.1的鐮孢毒素羧酸酯酶的值。

根據(jù)本發(fā)明的一個進(jìn)一步改造方案，所述多肽變體在至少一個從由66、199、302、377、394、424、430和463組成的組中選擇的位置處具有氨基酸置換，并且具有相比于具有seqidno.1的鐮孢毒素羧酸酯酶而言提高了至少6％的溫度穩(wěn)定性。借助于這樣的多肽變體，可能的是，在具有高的溫度的技術(shù)過程例如面糊制品和其他玉米產(chǎn)品(例如玉米粥、爆米花、玉米片、玉米面包或玉米粉圓餅)的生產(chǎn)，以及淀粉液化過程、糖化過程或發(fā)酵過程(例如制漿過程或發(fā)酵過程，特別是在生物乙醇制備中)期間裂解鐮孢毒素。

根據(jù)本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案，所述氨基酸置換選自由下列各項(xiàng)組成的組：66d、199i、302i、377v、394p、424a、430a和463d。借助于這樣的置換，可以使所述多肽變體的溫度穩(wěn)定性以及溫度活性相對于seqidno.1的鐮孢毒素羧酸酯酶而言提高至少3℃。

根據(jù)本發(fā)明的一個進(jìn)一步改造方案，所述多肽變體在該氨基酸序列的至少兩個，特別是三個從由下列各項(xiàng)組成的組中選擇的位置處具有氨基酸置換：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490，并且具有相比于seqidno.1的鐮孢毒素羧酸酯酶而言提高了至少15％的溫度穩(wěn)定性。

根據(jù)本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案，所述氨基酸置換選自由下列各項(xiàng)組成的組：10q、66d、144m、151r、199i、266s、267p、272h、275e、275a、280d、284t、286p、286r、293e、302i、360v、363t、364h、364l、365i、367h、371v、371m、372f、377v、389l、391v、394p、418a、419v、424a、424k、427v、429p、430a、436a、436s、440g、440s、443t、447a、453r、455s、456q、457t、462y、463d、464i、465h、465s、465g、469k、473a、478d、487n和490p。通過多個氨基酸的置換，已經(jīng)令人驚訝地顯示，大致加上了其對于溫度穩(wěn)定性的正面效應(yīng)，其中通過嵌入至少三個與原來在該序列中存在的氨基酸不同的氨基酸，溫度穩(wěn)定性提高了多于7℃。這樣的提高足以例如在生物乙醇制備中在處于大約55℃下的制漿停歇之時或者在處于大約65至70℃的中等溫度下的飼料?；畷r使用所述酶。

根據(jù)本發(fā)明的一個進(jìn)一步改造方案，所述多肽變體的氨基酸序列包含多個氨基酸置換的組合，其中所述位置組合選自由下列各項(xiàng)組成的組：66/199/302/394/424/430、66/199/302/377/394/424/430、66/199/302/377/394/424/430/463、66/144/199/302/360/372/377/394/424/430/443/463、199/302/377/394/424/430/463、66/199/302/377/394、66/199/302/364/377/394/424/430/463、66/199/302/377/394/424/430/463/465、66/199/302/377/394/424/430/440/463、66/199/302/377/394/424/430/447/463、66/199/302/377/394/418/424/430/463、66/199/302/377/394/424/436/430/463、66/199/302/364/377/394/424/430/463、66/199/302/377/394/424/430/463/490、66/199/302/377/394/424/430/463/469、66/199/302/377/389/394/424/430/463、66/199/302/377/394/424/430/463/465、66/199/302/377/394/424/430/463/464、66/199/302/377/394/424/430/463/465、66/199/302/377/394/424/430/440/463、66/199/302/377/394/424/430/457/463、66/199/302/377/394/424/430/436/463、66/199/302/363/371/377/394/424/430/463、66/199/302/377/394/424/430/447/453/463、66/199/302/377/394/424/430/456/462/463、66/199/302/377/394/419/424/427/430/463、66/199/302/365/377/394/424/430/463/487和66/199/302/371/377/394/424/430/463/487。如此經(jīng)置換的多肽變體具有相比于seqidno.1的鐮孢毒素羧酸酯酶而言提高了至少25％的溫度穩(wěn)定性。通過這樣的置換組合，可能的是，不僅提高了所述多肽變體的溫度穩(wěn)定性，而且還提高了所述多肽變體的溫度活性。

根據(jù)本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案，所述多肽變體的氨基酸序列選自由seqidno.2至29組成的組。這樣的多肽變體具有相比于seqidno.1的酶而言提高了至少11℃，優(yōu)選地至少13℃和更加優(yōu)選地至少15℃的溫度穩(wěn)定性，由此在具有高的溫度負(fù)荷的技術(shù)過程(例如在生物乙醇制備中處于65℃下的制漿停歇，或者處于大約75-80℃的溫度下的?；?期間或之后保證各自多肽的活性和因此鐮孢毒素的解毒。

根據(jù)本發(fā)明的一個進(jìn)一步改造方案，所述多肽變體的每一個氨基酸序列包含多個氨基酸置換的組合，其中所述位置組合選自由下列各項(xiàng)組成的組：66/99/302/364/377/389/394/419/424/427/430/447/463/465/469、66/199/302/377/389/394/419/424/427/430/447/463/465/469、66/199/302/363/364/371/377/389/394/419/424/427/430/447/463/464/465/469、66/199/302/363/371/377/389/394/419/424/427/430/447/463/464/465/469、66/199/302/364/367/371/377/389/394/418/419/424/427/430/436/440/447/463/464/465/469/490、66/199/302/367/371/377/389/394/418/419/424/427/430/436/440/447/463/464/465/469/490、66/199/302/363/367/371/377/394/424/430/463/490、66/199/302/377/394/418/419/424/427/430/436/440/447/463、66/199/302/377/389/394/424/430/457/463/464/465/469、66/199/302/363/371/377/389/394/419/424/427/430/440/447/457/463/464/469/490、66/199/302/377/394/424/430/463/447/490/469/465、66/199/302/377/394/424/430/463/490/469/465/419/427/371/487、66/199/302/371/377/394/419/424/427/430/447/453/463/465/469/487/490、66/199/302/367/371/377/389/394/418/419/424/427/429/430/436/440/447/457/463/464/465/469/490、66/199/302/371/377/389/394/419/424/427/430/436/447/453/456/462/463/465/469/490/487和66/199/302/367/371/377/389/394/418/419/424/427/429/430/436/440/447/453/456/457/462/463/464/465/469/487/490。這些多肽變體具有相比于seqidno.1的鐮孢毒素羧酸酯酶而言提高了至少40％的溫度穩(wěn)定性以及增高了的溫度活性，由此它們在許多需要更高溫度的方法中是可使用的。

根據(jù)本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案，所述多肽變體的氨基酸序列選自由seqidno.30至45組成的組。這樣的多肽變體具有至少18℃，優(yōu)選地至少22℃，和更優(yōu)選地至少27℃的相比于seqidno.1的酶而言提高的溫度穩(wěn)定性，由此在具有高的溫度負(fù)荷的技術(shù)過程(例如處于超過80℃，特別地超過85-90℃的溫度下的粒化)期間或之后保證所述多肽的活性和因此鐮孢毒素的解毒。特別地，在大約90℃的高的溫度下的?；诩仪莨I(yè)中具有重大意義，以便保證飼料中沙門氏菌負(fù)荷的充分降低。

名稱“保守突變”涉及氨基酸由被專業(yè)人員視為保守的(即具有類似的特殊特性的)其他氨基酸來進(jìn)行的置換，或者說該氨基酸的特性仍然是得到保存的，即得以保留的。氨基酸的特殊特性為例如其大小、極性、疏水性、電荷或pk值?？梢詫被峄谄涮匦詣澐譃榻M，并且所述組可以以維恩圖(venndiagram)來描繪。來自相同組和優(yōu)選地來自相同亞組的氨基酸可以相互置換。依據(jù)疏水性、極性和大小這些特性把氨基酸劃分為組和亞組可以從taylorw.r.(1986)中獲知?！氨Ｊ赝蛔儭笔侵咐鐚⒁粋€酸性氨基酸置換成另一個酸性氨基酸，將一個堿性氨基酸置換成另一個堿性氨基酸，將一個極性氨基酸置換成另一個極性氨基酸，等等。特別地，所述多肽變體另外還可以在至少一個位置處包含至少一個保守氨基酸置換，其中所述保守氨基酸置換選自由下列置換組成的組：g至a，a至g/s，v至i/l/a/t/s，i至v/l/m，l至i/m/v，m至l/i/v，p至a/s/n，f至y/w/h，y至f/w/h，w至y/f/h，r至k/e/d，k至r/e/d，h至q/n/s，d至n/e/k/r/q，e至q/d/k/r/n，s至t/a，t至s/v/a，c至s/t/a，n至d/q/h/s，和q至e/n/h/k/r。

如果在根據(jù)本發(fā)明的多肽變體中在確定位置處的置換導(dǎo)致例如極性氨基酸(例如asp)被疏水氨基酸(例如ala)替代，那么“保守突變”也是指所有在所述位置處導(dǎo)致另一個疏水氨基酸(例如甘氨酸、亮氨酸、苯丙氨酸、纈氨酸)的突變。在此也包括這些包含備選的保守突變的進(jìn)一步的多肽變體。

此外，本發(fā)明還旨在提供編碼seqidno.1的鐮孢毒素羧酸酯酶的裂解鐮孢毒素的多肽變體的多核苷酸以供使用，用所述多肽變體可成功實(shí)現(xiàn)在不依賴于氧且無輔因子的情況下將至少一種鐮孢毒素裂解為無毒或低毒的產(chǎn)物，并且所述多肽變體具有相比于seqidno.1的鐮孢毒素羧酸酯酶而言提高的溫度穩(wěn)定性。

為了解決該任務(wù)，本發(fā)明的特征在于，所述多核苷酸具有編碼具有seqidno.1的氨基酸序列的鐮孢毒素羧酸酯酶的裂解鐮孢毒素的多肽變體的核苷酸序列，其中所述多肽變體具有與具有seqidno.1的氨基酸序列有著至少70％的序列同一性的氨基酸序列，并且所述多肽變體在至少一個從由下列各項(xiàng)組成的組中選擇的位置處具有氨基酸置換：10、33、66、107、140、144、149、151、157、199、266、267、270、272、275、280、284、286、293、302、312、329、332、360、363、364、365、367、371、372、377、389、391、394、418、419、424、427、429、430、436、440、443、447、453、455、456、457、462、463、464、465、469、473、478、487和490，然而特別地具有下列置換：10q、33e、66d、107e、140p、144m、149f、151r、157y、199i、266s、267p、270f、272h、275e、275a、280d、280p、284t、284p、286p、286r、293e、302i、312f、329f、332e、360v、363t、364h、364l、365i、367h、371v、371m、372f、377v、389l、391v、394p、418a、419v、424a、424k、427v、429p、430a、436a、436s、440g、440s、443t、447a、453r、455s、456q、457t、462y、463d、464i、465h、465s、465g、469k、473a、478d、487n和490p或者其組合。用這樣的分離的多核苷酸，可以借助于表達(dá)載體來制備用于產(chǎn)生根據(jù)本發(fā)明的多肽變體的轉(zhuǎn)基因宿主細(xì)胞。

名稱“表達(dá)載體”涉及能夠在體內(nèi)或在體外表達(dá)基因的dna構(gòu)建體。特別地，該名稱涵蓋這樣的dna構(gòu)建體，其適合于將編碼所述多肽的核苷酸序列轉(zhuǎn)移到宿主細(xì)胞中，從而使得所述核苷酸序列整合到基因組中或者游離地存在于染色體外空間中，并且在細(xì)胞內(nèi)表達(dá)編碼所述多肽的核苷酸序列和任選地從細(xì)胞中提取出所述多肽。

名稱“宿主細(xì)胞”涉及所有這樣的細(xì)胞，其要么包含待表達(dá)的核苷酸序列，要么包含表達(dá)載體，并且能夠制備根據(jù)本發(fā)明的酶或多肽。特別地，該名稱涵蓋原核細(xì)胞和/或真核細(xì)胞，優(yōu)選地巴斯德畢赤酵母(pichiapastoris)，大腸桿菌(e.coli)，枯草芽孢桿菌(bacillussubtilis)，鏈霉菌屬(streptomyces)，漢遜酵母屬(hansenula)，木霉屬(trichoderma)，乳桿菌屬(lactobacillus)，曲霉屬(aspergillus)，植物細(xì)胞，和/或芽孢桿菌屬(bacillus)、木霉屬或曲霉屬的孢子。在此所使用的名稱“巴斯德畢赤酵母(pichiapastoris)”是名稱“komagataellapastoris”的同義詞，其中巴斯德畢赤酵母(pichiapastoris)是較老的名稱而komagataellapastoris是系統(tǒng)分類學(xué)上較新的名稱(yamada等人,1995)。

此外，本發(fā)明還旨在提供裂解鐮孢毒素的添加劑以供使用，所述添加劑包含至少一種具有seqidno.46的氨基酸序列的鐮孢毒素羧酸酯酶的裂解鐮孢毒素的多肽變體，其中各自多肽變體在不依賴于氧且無輔因子的情況下將至少一種鐮孢毒素裂解為無毒或低毒的產(chǎn)物并且具有相比于seqidno.46的鐮孢毒素羧酸酯酶而言提高的溫度穩(wěn)定性。

為了解決該任務(wù)，本發(fā)明的特征在于，所述裂解鐮孢毒素的添加劑包含至少一種根據(jù)本發(fā)明的鐮孢毒素羧酸酯酶的多肽變體，和任選地，至少一種助劑。通過將這樣的添加劑添加至被鐮孢毒素污染的飼料，可以成功實(shí)現(xiàn)使鐮孢毒素解毒，這可以顯著地通過在用所述添加劑飼喂的受試者的血漿和/或腎臟和/或肺和/或肝臟中二氫鞘氨醇與鞘氨醇的比例的降低來進(jìn)行測量。

在動物的不同器官中以及在動物的血漿中二氫鞘氨醇與鞘氨醇的比例是關(guān)于鐮孢毒素(特別是fb1)的毒性效應(yīng)的普遍接受且靈敏的生物標(biāo)志物。由鐮孢毒素引起的鞘脂代謝紊亂尤其與馬的腦病或豬中的肺水腫有關(guān)。作為生物標(biāo)志物的二氫鞘氨醇與鞘氨醇的比例的重要性以及其分析測量描述在grenier等人(biochem.pharmaceuticals,第83卷,(2012),第1465-1473頁)中以及描述在efsajournal(2014,12(5):3667)中。

根據(jù)本發(fā)明的一個進(jìn)一步改造方案，如此地來形成所述添加劑，從而使得所述助劑選自由下列各項(xiàng)組成的組：惰性載體、維生素、礦物質(zhì)、植物源性物質(zhì)、酶和其他用于使真菌毒素解毒的組分，例如真菌毒素降解酶，特別是黃曲霉毒素氧化酶、麥角胺水解酶、麥角胺酰胺酶、玉米赤霉烯酮酯酶、玉米赤霉烯酮內(nèi)酯酶、玉米赤霉烯酮水解酶、赭曲毒素酰胺酶、串珠鐮孢菌素氨基轉(zhuǎn)移酶、氨基多元醇氨基氧化酶、脫氧瓜萎鐮菌醇環(huán)氧化物水解酶、脫氧瓜萎鐮菌醇脫氫酶、脫氧瓜萎鐮菌醇氧化酶、單端孢霉烯脫氫酶、單端孢霉烯氧化酶；降解真菌毒素的微生物；和結(jié)合真菌毒素的物質(zhì)，例如微生物細(xì)胞壁或無機(jī)材料例如膨潤土。通過使用這樣的添加劑，可以例如在飼料或食物中確保，所任選地包含的鐮孢毒素的量安全可靠地被裂解特別是解毒至如此程度，即對于攝入了這些飼料或食物的受試者的機(jī)體的有害效應(yīng)不存在。

本發(fā)明的另外的使用領(lǐng)域?yàn)檫@樣的添加劑，其除了包含至少一種根據(jù)本發(fā)明的多肽變體外，還包含至少一種酶，所述酶例如參與蛋白質(zhì)的降解，例如蛋白酶，或者所述酶參與淀粉或纖維或脂肪或糖原的代謝，例如淀粉酶、纖維素酶或葡聚糖酶，以及例如水解酶、脂解酶、甘露糖苷酶、氧化酶、氧化還原酶、植酸酶或木聚糖酶。

此外，本發(fā)明還旨在根據(jù)本發(fā)明的添加劑用于裂解在食物或飼料中的至少一種鐮孢毒素的用途，所述食物或飼料特別地是經(jīng)?；?，所述飼料特別地為用于豬、家禽、牛、馬、魚類或水產(chǎn)養(yǎng)殖的飼料。作為食物或飼料，可以使用所有適合于人或動物營養(yǎng)(特別地也適合于家畜、綿羊或山羊)的食物或飼料，特別地也使用干酒槽。

此外，本發(fā)明還旨在根據(jù)本發(fā)明的添加劑用于在用于制備或加工食物或飼料的過程中裂解至少一種鐮孢毒素的用途，所述過程特別地在至少50℃的溫度下進(jìn)行。通過所述添加劑的這樣的用途，保證了鐮孢毒素的解毒，例如在其中在高的溫度下的處理是重要的食品技術(shù)過程期間，例如在玉米或谷物的加工中，在淀粉液化過程中，在糖化過程中，或在發(fā)酵過程(例如制漿過程或發(fā)酵過程，特別是在生物乙醇制備中)中。由此可以確保，在來自于此類方法的產(chǎn)品(例如飼料顆粒、面糊制品、玉米粥、爆米花、玉米片、玉米面包、玉米粉圓餅、干酒糟或淀粉)中沒有重大的(特別地有損于健康的)量的鐮孢毒素保持完整。

此外，本發(fā)明還旨在至少一種根據(jù)本發(fā)明的多肽變體或者至少一種根據(jù)本發(fā)明的添加劑在制備用于預(yù)防和/或治療鐮孢毒素真菌毒素中毒的制劑中的用途。通過多肽變體或添加劑的這樣的用途，在預(yù)防的情況下可能的是，盡管經(jīng)口攝入了鐮孢毒素，但是將人或動物的健康狀況基本上保持在這樣的水平，該水平相應(yīng)于沒有經(jīng)口攝入鐮孢毒素或經(jīng)口攝入了減少的鐮孢毒素的水平。在治療鐮孢毒素真菌毒素中毒的情況下，可能的是，緩解此類疾病的癥狀和特別地顯著改善在器官和/或血漿中二氫鞘氨醇與鞘氨醇的比例。此外，通過這樣的用途，還可能的是，改善有用動物的生產(chǎn)能力，特別是飼料利用率和體重增加，并且降低死亡率。

此外，本發(fā)明還旨在提供用于以酶促方式裂解至少一種鐮孢毒素的方法以供使用，采用所述方法可以用多肽將至少一種鐮孢毒素不依賴于氧地、特異性地、安全地和可靠地以水解方式裂解為無毒或低毒的產(chǎn)物，其中所述以水解方式的裂解要么在熱處理期間要么在熱處理之后進(jìn)行。

為了解決該任務(wù)，如此地來施行所述方法，從而通過根據(jù)本發(fā)明的多肽變體或根據(jù)本發(fā)明的添加劑從鐮孢毒素中以水解方式裂解出至少一個丙三羧酸。在此，將所述至少一種鐮孢毒素與至少一種根據(jù)本發(fā)明的多肽變體或至少一種根據(jù)本發(fā)明的添加劑相混合，通過至少一種多肽變體從至少一種鐮孢毒素中以水解方式裂解出至少一個丙三羧酸，由此使所述鐮孢毒素解毒，其中使由各自多肽變體和鐮孢毒素組成的混合物經(jīng)歷至少50℃，優(yōu)選地至少70℃的熱處理，并且所述以水解方式的裂解要么在所述熱處理期間要么在所述熱處理之后進(jìn)行。

在本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案中，如此地來施行所述方法，從而將所述多肽變體或所述添加劑與被至少一種鐮孢毒素污染的飼料或食物相混合，并且進(jìn)行熱處理(任選地通過?；^程)。由此確保，一旦由所述多肽變體和所述鐮孢毒素組成的混合物與濕氣相接觸，在被污染的和任選地經(jīng)粒化的飼料或食物中所包含的鐮孢毒素就被裂解。在濕潤的飼料或食物例如麥芽漿或糊漿的情況下，鐮孢毒素的水解在經(jīng)口攝入之前在該濕潤的飼料或食物中發(fā)生。由此達(dá)到，最大程度地消除或至少降低鐮孢毒素對于人和動物的有害效應(yīng)。在此，濕氣是指水或含水液體的存在，其中例如唾液或其他在消化道中存在的液體也屬于該范疇。消化道被定義為口腔、咽(咽喉)、食管和胃腸道或者其等價物，其中在動物中可以存在不同的名稱或者個別成分可以不存在于動物的消化道中。

在本發(fā)明的一個優(yōu)選的進(jìn)一步改造方案中，如此地來施行所述方法，以5u至500u，優(yōu)選地10u至300u，和更優(yōu)選地15u至100u/千克飼料或食物的濃度范圍使用所述多肽變體。通過添加這些量的多肽變體，可成功實(shí)現(xiàn)，取決于鐮孢毒素的濃度，將在食物或飼料中(特別地在干酒糟中)的鐮孢毒素裂解并因此使其解毒至如此程度，即至少70％，優(yōu)選地至少80％，特別地至少90％的至少一種鐮孢毒素被裂解。

如果沒有明確地另外說明，那么單數(shù)名稱例如“ein”、“eine”、“das”、“die”或“der”被理解為示例性的并且包括許多可能性。例如，如果涉及“基因”、“酶”或“細(xì)胞”，那么總是意圖包括復(fù)數(shù)。

下面將依據(jù)實(shí)施例更詳細(xì)地闡明本發(fā)明。

多肽變體

實(shí)施例1：編碼裂解鐮孢毒素的多肽的多核苷酸的修飾、克隆和表達(dá)

按照說明書，使用“quick-changesite-directedmutagenesiskits”(stratagene)，借助于pcr，通過核苷酸序列的突變來進(jìn)行氨基酸置換、插入或缺失。備選地，為此還合成制備了完整的核苷酸序列(geneart)。借助于標(biāo)準(zhǔn)方法，將借助于pcr誘變而產(chǎn)生的或從geneart購進(jìn)的核苷酸序列整合到用于在大腸桿菌(e.coli)或巴斯德畢赤酵母(p.pastoris)中進(jìn)行表達(dá)的表達(dá)載體之中，轉(zhuǎn)化到大腸桿菌或巴斯德畢赤酵母中，以及在大腸桿菌或巴斯德畢赤酵母中進(jìn)行表達(dá)(j.m.cregg,pichiaprotocols,第二版,isbn-10:1588294293、2007；j.sambrook等人,2012,molecularcloning,alaboratorymanual,第4版,coldspringharbor)，其中也可以為該任務(wù)考慮任何其他合適的宿主細(xì)胞。

名稱“表達(dá)載體”涉及能夠在體內(nèi)或在體外表達(dá)基因的dna構(gòu)建體。特別地，該名稱涵蓋這樣的dna構(gòu)建體，其適合于將編碼所述多肽的核苷酸序列轉(zhuǎn)移到宿主細(xì)胞中，以便在那里整合到基因組中或者游離地存在于染色體外空間中，并且在細(xì)胞內(nèi)表達(dá)編碼所述多肽的核苷酸序列和任選地從細(xì)胞中提取出所述多肽。名稱“宿主細(xì)胞”涉及所有這樣的細(xì)胞，其要么包含待表達(dá)的核苷酸序列，要么包含表達(dá)載體，并且能夠制備根據(jù)本發(fā)明的酶或多肽。特別地，該名稱涵蓋原核細(xì)胞和/或真核細(xì)胞，優(yōu)選地巴斯德畢赤酵母(p.pastoris)，大腸桿菌(e.coli)，枯草芽孢桿菌(bacillussubtilis)，鏈霉菌屬(streptomyces)，漢遜酵母屬(hansenula)，木霉屬(trichoderma)，乳桿菌屬(lactobacillus)，曲霉屬(aspergillus)，植物細(xì)胞，和/或芽孢桿菌屬(bacillus)、木霉屬或曲霉屬的孢子?？紤]將可溶的細(xì)胞裂解物(在大腸桿菌的情況下)或者培養(yǎng)物上清液(在巴斯德畢赤酵母的情況下)用于測定所述多肽變體的催化特性。

實(shí)施例2：降解鐮孢毒素的多肽的催化活性和比活性的測定

借助于標(biāo)準(zhǔn)方法將編碼降解鐮孢毒素的多肽變體的相應(yīng)基因克隆到大腸桿菌中，在細(xì)胞內(nèi)進(jìn)行表達(dá)，并隨后借助于超聲波進(jìn)行裂解并進(jìn)行離心。將清澈的上清液用具有0.1mg/ml牛血清白蛋白的20mmtris-hcl緩沖液(ph8.0)進(jìn)行稀釋(大約10^-3至10^-5)，并且如此地用于fb1降解批料中，從而通過該多肽將10％至90％的在降解批料中所包含的fb1量降解。

為了測定催化酶活性，進(jìn)行用于以水解方式裂解串珠鐮孢菌素b1(fb1)的試驗(yàn)，其中使試驗(yàn)批料在具有0.1mg/ml牛血清白蛋白的20mmtris-hcl緩沖液(ph8.0)中和在30℃的溫度下進(jìn)行30分鐘。此外，所述批料還包含100μμfb1的底物濃度(biopurereferenzsubstanzengmbhtulln,austria,brm001007)和待測試的多肽。在30分鐘的溫育后，將所述批料在99℃下熱滅活5分鐘以終止反應(yīng)。

為了測定飼料樣品的酶促活性，必須在所述試驗(yàn)之前從飼料樣品中提取出轉(zhuǎn)化鐮孢毒素的多肽變體。為此將10g飼料溶解在100ml的具有0.1mg/ml牛血清白蛋白的20mmtris-hcl緩沖液(ph8.0)中，并在20℃和150rpm下?lián)u動1小時。然后，將樣品在4000g下離心15分鐘，并且將清澈的上清液根據(jù)需要進(jìn)行稀釋(10^-2至10^-3)并用于fb1批料中。

按照heinl等人(j.ofbiotechnology,2010,145,第120-129頁,2.6.3.“l(fā)iquidchromatography-massspectrometry”)的方法，借助于lc-ms(液相色譜法-質(zhì)譜法)來進(jìn)行fb1的定量。為此，考慮用fb1標(biāo)準(zhǔn)物進(jìn)行校準(zhǔn)，所述fb1標(biāo)準(zhǔn)物還包含完整的經(jīng)¹³c標(biāo)記的內(nèi)部fb1標(biāo)準(zhǔn)物(biopurereferenzsubstanzengmbhtulln,austria)。與heinl等人(2010)不同，僅測量了fb1的降解，以便測定所使用的多肽溶液的催化酶活性。所使用的多肽溶液的催化酶活性以“單位/ml”來給出，其中將一個“單位”定義為在該試驗(yàn)中在上述反應(yīng)條件下1μmolfb1/分鐘的減少。

為了測定比活性，借助于定量western印跡或elisa來測定酶濃度。通過將活性(單位)與所使用的酶量相關(guān)聯(lián)來計算比酶活性，并且以“單位/mg”來給出。

實(shí)施例3：降解鐮孢毒素的多肽的溫度穩(wěn)定性

如在實(shí)施例1和2中所描述的那樣，進(jìn)行降解鐮孢毒素的多肽的表達(dá)和定量。在活性測定之前，將細(xì)胞裂解物的量分為多個部分(每個大約60μl)。使2至10個部分在商用pcr-循環(huán)儀(例如eppendorfmastercyclergradient)中經(jīng)歷熱處理5分鐘，其中將每個部分在不同的溫度下進(jìn)行溫育。同時，將細(xì)胞裂解物的另一部分，100％-對照，在冰上進(jìn)行溫育。在熱處理之后，將所有樣品/試驗(yàn)批料在10℃下溫育一分鐘以使溫度相稱。如在實(shí)施例2中所描述的那樣，測定經(jīng)熱處理的樣品以及100％-對照的酶促活性。在熱處理后的殘留活性被稱為剩余活性。將相比于未經(jīng)熱處理的100％-對照而言的剩余活性為50％時所處的溫度縮寫為t(50％)，并且其是關(guān)于多肽的溫度穩(wěn)定性的量度。

相比于具有seqidno.46或seqidno.1的多肽而言多肽變體的t(50％)(其以攝氏度給出)的增加是關(guān)于提高的溫度穩(wěn)定性的量度。t(50％)-值的增加可以以℃，但也可以以相對于親本多肽的t(50％)-值而言的百分比值來給出。下面的例子將會用于進(jìn)行闡明：如果親本酶在冰上溫育5分鐘后具有50u/ml的催化活性并且在48℃下溫育5分鐘后具有25u/ml的催化活性，那么t(50％)為48℃。如果多肽變體具有51℃的t(50％)，那么溫度穩(wěn)定性(t(50％))的相對提高為6.25。這從所述兩個t(50％)-值的3℃的差除以親本起始酶的48℃的t(50％)-值再乘以100而得出。

為了測定溫度穩(wěn)定性，也可以考慮比活性來代替催化活性。

溫度穩(wěn)定性的測定也可以借助于備選的酶測定法來進(jìn)行，甚至無需測定fb1轉(zhuǎn)化的催化活性。在此重要的是，在批料中無論如何一定要使用相同量的經(jīng)熱處理的多肽和100％-對照，這例如通過使用相同體積的細(xì)胞裂解物來提供。

為了測定溫度穩(wěn)定性，也可以考慮酶促降解試驗(yàn)的測量信號(例如ms信號、消光等)來代替催化活性。如果所述測量信號與酶促活性(例如，經(jīng)轉(zhuǎn)化的fb1的峰面積)成正比，那么多肽的t(50％)-值為這樣的溫度，在所述溫度下，經(jīng)熱處理的多肽的測量信號值具有該多肽的100％-對照的測量信號值的50％。

測得具有seqidno.46的多肽的溫度穩(wěn)定性(t(50％))為42℃，具有seqidno.1的多肽的溫度穩(wěn)定性為45℃。因此，通過縮短n-末端序列可以達(dá)到的溫度穩(wěn)定性的相對提高為大約7％。此外，還可以查明具有seqidno.1的多肽相比于具有seqidno.46的親本多肽而言的提高的酶促活性。

具有seqidno.1的多肽的溫度穩(wěn)定性可以通過單個氨基酸的有針對性的置換而進(jìn)一步得到提高。這些多肽變體相比于具有seqidno.1的親本多肽而言的溫度穩(wěn)定性的相對提高呈現(xiàn)在表1中。

表1：多肽變體的修飾以及其相比于具有seqidno.1的親本酶而言的以百分比表示的溫度穩(wěn)定性的相對提高

實(shí)施例4：降解鐮孢毒素的多肽的依賴于溫度的活性(溫度活性)

在試驗(yàn)進(jìn)行之前純化待就其依賴于溫度的活性進(jìn)行測試的、降解鐮孢毒素的多肽變體。為此，借助于陰離子交換色譜法和隨后的大小排阻色譜法以兩步驟過程從發(fā)酵上清液中純化出多肽變體。將多肽變體調(diào)整至1mg/ml的濃度，并且在1ml的反應(yīng)體積中以10^-5-10^-6的稀釋度用于反應(yīng)批料中。如在實(shí)施例3中所描述的試驗(yàn)?zāi)菢?，通過fb1水解和隨后的借助于lc-ms的fb1的定量來進(jìn)行活性測定，其中所述試驗(yàn)在各種不同的溫度下進(jìn)行。用兩個加熱板(eppendorf,thermomixer)在溫度10℃、20℃、30℃、35℃、40℃、45℃、50℃、55℃、60℃、65℃和70℃下進(jìn)行溫育。在熱作用開始后30分鐘，各取出100μl的反應(yīng)批料并在99℃下熱滅活5分鐘。在此，在加熱板中于30℃下進(jìn)行的試驗(yàn)用作100％-對照。在表2中列舉了示例性結(jié)果。表2：降解鐮孢毒素的多肽的依賴于溫度的活性。<loq的值低于檢測極限(定量水平(levelofquantification)：<0.15u/l，在試驗(yàn)批料中)

實(shí)施例5：降解鐮孢毒素的多肽的?；€(wěn)定性的測定

借助于標(biāo)準(zhǔn)方法，將所選擇的多肽變體克隆到巴斯德畢赤酵母中，在受控的有氧條件下在生物反應(yīng)器中進(jìn)行表達(dá)，并分泌到細(xì)胞外。將清澈的上清液與生物質(zhì)分開，向其摻入載體物質(zhì)(麥芽糖糊精)，并且借助于噴霧干燥設(shè)備將其加工成可?；姆勰?。將以粉末形式存在的降解鐮孢毒素的多肽變體各自以100u/kg的相同濃度與仔豬飼養(yǎng)飼料相混合，并且在受控的過程中加工成飼料顆粒。在顆粒化過程期間，將飼料以單個批次在精確確定的溫度(75-95℃，以5℃的間距)下借助于熱蒸汽進(jìn)行潤濕并加熱。在該準(zhǔn)備階段后，進(jìn)行真正的?；^程。如在實(shí)施例2中所描述的那樣，測定在所述顆粒中所包含的降解鐮孢毒素的多肽變體的剩余活性，其中未經(jīng)粒化的具有各自的降解鐮孢毒素的多肽變體的飼料用作100％-對照。因此，在顆?；^程后殘留的酶活性被定義為剩余活性。所述值在表3中給出。

表3：降解鐮孢毒素的多肽的粒化溫度和剩余活性。<loq的值低于檢測極限(定量水平：<0.15u/l，在試驗(yàn)批料中)

序列表

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<120>裂解鐮孢毒素的多肽變體、包含所述多肽變體的添加劑和所述添加劑的用途，以及用于裂解鐮孢毒素的方法

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prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargphegluarg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>4

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>4

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>5

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>5

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglyphemet

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleuvalglyaspserglnpheasnasngly

355360365

ilegluleupheseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpthrserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>6

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>6

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyasngluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>7

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>7

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleugluglyglyalaglyalaseraspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargphegluarg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>8

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>8

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserhispheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>9

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>9

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

hisglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>10

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>10

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetserglyalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>11

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>11

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>12

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>12

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valalametpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>13

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>13

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualaalaglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>14

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>14

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserleupheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>15

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>15

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>16

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>16

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>17

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>17

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>18

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>18

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

serglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>19

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>19

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheaspile

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>20

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>20

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

glyglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>21

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>21

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetserseralatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>22

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>22

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysthrhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>23

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>23

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualaserglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>24

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>24

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspthrglnpheasnasngly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>25

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>25

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspargglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>26

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>26

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrglnserhistrpproargtyrasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>27

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>27

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>28

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>28

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnileasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserasnalacysglnproserlys

485490

<210>29

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>29

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ileglumetleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserasnalacysglnproserlys

485490

<210>30

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>30

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserleupheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>31

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>31

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>32

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>32

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspthrleupheasnasngly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheaspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>33

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>33

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspthrglnpheasnasngly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheaspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>34

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>34

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserleupheasnhisgly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valalavalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualaalaglumetserseralatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheaspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>35

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>35

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnhisgly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valalavalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualaalaglumetserseralatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheaspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>36

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>36

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspthrglnpheasnhisgly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>37

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>37

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valalavalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualaalaglumetserseralatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

argglygluilemetthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>38

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>38

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysthrhistrpproargpheaspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysglnproserlys

485490

<210>39

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>39

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspthrglnpheasnasngly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetserseralatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysthrhistrpproargpheaspile

450455460

argglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>40

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>40

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ilegluleuleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valsermetpheglyserleualaglyglyalaglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>41

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>41

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ileglumetleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealavalhis

435440445

glyvalproaspglnglythrlysserhistrpproargpheasparg

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserasnalacysproproserlys

485490

<210>42

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>42

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ileglumetleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargphethrglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualathrglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspargglythrlysserhistrpproargpheasparg

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserasnalacysproproserlys

485490

<210>43

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>43

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnhisgly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valalavalpheglyserleualaglyglyvalglyproalaaspile

420425430

lysleualaalaglumetserseralatrpvalserphealaalahis

435440445

glyvalproaspglnglythrlysthrhistrpproargpheaspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserlysalacysproproserlys

485490

<210>44

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>44

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnasngly

355360365

ileglumetleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valservalpheglyserleualaglyglyvalglyalaalaaspile

420425430

lysleualaalaglumetseralaalatrpvalserphealaalahis

435440445

glyvalproaspargglythrglnserhistrpproargtyrasparg

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserasnalacysproproserlys

485490

<210>45

<211>493

<212>prt

<213>人工序列

<220>

<223>seqidno.1的多肽變體

<400>45

glnthraspaspprolysleuvalarghisthrglnserglyalaval

151015

gluglyvalgluglyaspvalgluthrpheleuglyilepropheala

202530

alaproprovalglyaspleuargtrpargproproalaproproarg

354045

alatrpalaglythrargaspglyargargphealaproaspcysile

505560

glyaspgluargleuarggluglyserargalaalaglythrserglu

65707580

aspcysleutyrleuasniletrpserprolysglnvalglylysgly

859095

glyleuprovalmetiletrpvaltyrglyglyglypheserglygly

100105110

serglyalavalprotyrtyraspglyseralaleualaglnlysgly

115120125

valvalvalvalthrpheasntyrargalaglyileleuglypheleu

130135140

alahisproalaleuserlysgluserproasnglyvalserglyasn

145150155160

tyrglyleuleuaspmetleualaalaphelystrpvalglnasnasn

165170175

ileargglupheglyglyaspproasnargvalthrvalpheglyglu

180185190

seralaglyalaseralaileglyleuleuleuthrserproleuser

195200205

gluseralapheasnglnalaileleuglnserproglyleualaarg

210215220

proleualathrleusergluserglualaasnglyleugluleugly

225230235240

alaaspileseralaleuargargalaaspalaglygluleuthrlys

245250255

ilealaglnserargileprometserargglnphethrlysproarg

260265270

prometglyproileleuaspglytyrvalleuargthrleuaspval

275280285

aspalaphealalysglyalaphearglysileprovalilevalgly

290295300

glyasnalaaspgluglyargalaphethraspargleuprovallys

305310315320

thrvalleuglutyrargalatyrleuthrgluglnpheglyaspglu

325330335

alaaspalatrpgluargcystyrproalaasnseraspalaaspval

340345350

proalaalavalalaargleupheglyaspserglnpheasnhisgly

355360365

ilegluvalleuseralaalaphevallystrpargthrproleutrp

370375380

argtyrargpheleuglyileproglyproglyargargproalathr

385390395400

hisglyaspgluileprotyrvalphealaasnleuglyproserser

405410415

valalavalpheglyserleualaglyglyvalglyproalaaspile

420425430

lysleualaalaglumetserseralatrpvalserphealaalahis

435440445

glyvalproaspargglythrglnthrhistrpproargtyraspile

450455460

serglygluilelysthrpheglyserglnvalglyserglyglugly

465470475480

leuglyvalserproserasnalacysproproserlys

485490

<210>46

<211>540

<212>prt

<213>sphingopyxissp.

<400>46

vallysgluhisglncysargglyglyargalaserproalaalapro

151015

alathrtrpleualaargileservalserargglyalaseralaile

202530

alatrpthrphemetleuglyalathralaileprovalalaalagln

354045

thraspaspprolysleuvalarghisthrglnserglyalavalglu

505560

glyvalgluglyaspvalgluthrpheleuglyileprophealaala

65707580

proprovalglyaspleuargtrpargproproalaproproargala

859095

trpalaglythrargaspglyargargphealaproaspcysilegly

100105110

asngluargleuarggluglyserargalaalaglythrsergluasp

115120125

cysleutyrleuasniletrpserprolysglnvalglylysglygly

130135140

leuprovalmetiletrpvaltyrglyglyglypheserglyglyser

145150155160

glyalavalprotyrtyraspglyseralaleualaglnlysglyval

165170175

valvalvalthrpheasntyrargalaglyileleuglypheleuala

180185190

hisproalaleuserlysgluserproasnglyvalserglyasntyr

195200205

glyleuleuaspmetleualaalaphelystrpvalglnasnasnile

210215220

argglupheglyglyaspproasnargvalthrvalpheglygluser

225230235240

alaglyalaseralaleuglyleuleuleuthrserproleuserglu

245250255

seralapheasnglnalaileleuglnserproglyleualaargpro

260265270

leualathrleusergluserglualaasnglyleugluleuglyala

275280285

aspileseralaleuargargalaaspalaglygluleuthrlysile

290295300

alaglnserargileprometserargglnphethrlysproargpro

305310315320

metglyproileleuaspglytyrvalleuargthrleuaspvalasp

325330335

alaphealalysglyalaphearglysileprovalleuvalglygly

340345350

asnalaaspgluglyargalaphethraspargleuprovallysthr

355360365

valleuglutyrargalatyrleuthrgluglnpheglyaspgluala

370375380

aspalatrpgluargcystyrproalaasnseraspalaaspvalpro

385390395400

alaalavalalaargleupheglyaspserglnpheasnasnglyile

405410415

gluleuleuseralaalaphealalystrpargthrproleutrparg

420425430

tyrargphethrglyileproglyalaglyargargproalathrhis

435440445

glyaspgluileprotyrvalphealaasnleuglyproserserval

450455460

sermetpheglyserleugluglyglyalaglyalaseraspilelys

465470475480

leualathrglumetseralaalatrpvalserphealavalhisgly

485490495

valproaspglnglythrlysserhistrpproargphegluargarg

500505510

glygluilemetthrpheglyserglnvalglyserglygluglyleu

515520525

glyvalserproserlysalacysglnproserlys

530535540