CN111363735A - β-甘露聚糖酶耐热突变体、重组菌及其应用 - Google Patents

β-甘露聚糖酶耐热突变体、重组菌及其应用 Download PDF

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CN111363735A
CN111363735A CN202010275102.XA CN202010275102A CN111363735A CN 111363735 A CN111363735 A CN 111363735A CN 202010275102 A CN202010275102 A CN 202010275102A CN 111363735 A CN111363735 A CN 111363735A
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牟海津
刘哲民
宁晨
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Ocean University of China
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Abstract

本发明涉及β‑甘露聚糖酶耐热突变体、重组菌及其应用，属于基因工程和酶工程领域，本发明采用蛋白质表面电荷优化策略，理性设计并筛选突变位点，将白曲霉来源的β‑甘露聚糖酶ManAK进行改良，再通过高通量筛选方法挑选正向突变，最终获得热稳定性提高的23株β‑甘露聚糖酶突变体。本发明获得的β‑甘露聚糖酶突变体耐热性是原始基因的1.2～3.5倍，耐热性均有显著提高，有利于实现其在商业中的应用，从而降低生产成本。

Description

β-甘露聚糖酶耐热突变体、重组菌及其应用
技术领域
本发明属于基因工程和酶工程领域，具体涉及一类β-甘露聚糖酶耐热突变体、重组菌及其应用。
背景技术
甘露聚糖是半纤维素的第二大组分，由吡喃甘露糖通过β-1,4糖苷键连接而成。甘露聚糖的彻底降解需要甘露聚糖酶、甘露糖苷酶、葡萄糖苷酶、半乳糖苷酶和脱乙酰酯化酶的协同作用。其中，甘露聚糖酶为降解过程的关键酶。
β-甘露聚糖酶(EC 3.2.1.78)能通过催化β-1,4甘露糖苷键将甘露聚糖水解为甘露糖和甘露低聚糖。该酶作为一种重要的工业用酶制剂，被广泛应用于动物饲料、食品加工、医药、纺织、生物漂白以及石油等可再生能源领域。其中一些应用领域，特别是饲料制粒、可再生能源生产等过程需要进行高温处理，这就要求所开发的β-甘露聚糖酶具有良好的耐热性能。耐热酶的最适作用温度在60℃以上，较高的反应温度，降低了杂菌的污染，减少了杂菌代谢产物的干扰，可提高酶的催化能力和催化效率，极大降低生产成本。目前，市面上存在的β-甘露聚糖酶耐热性往往达不到工业要求,导致其在生产运输过程中生物活性不稳定，严重限制该酶应用。因此，开发具有优良耐热性的β-甘露聚糖酶具有重要工业意义和经济价值。
发明内容
本发明要解决的技术问题是提供一种耐热性提高的β-甘露聚糖酶突变体。本发明通过理性设计，采用蛋白质表面电荷优化策略，通过筛选设计位点，对白曲霉来源的β-甘露聚糖酶ManAK(氨基酸序列SEQIDNO：1，核苷酸序列SEQ IDNO：2)进行改良，得到热稳定性提高的β-甘露聚糖酶单位点突变体。本发明的另一目的是提供了包含上述突变的β-甘露聚糖酶基因的重组载体。将突变得到的β-甘露聚糖酶基因插入到表达载体合适位点，使其核苷酸序列可操作的与表达调控序列相连接，得到重组酵母表达质粒。本发明还提供了包含上述β-甘露聚糖酶基因的重组菌株，所述菌株为酵母菌。
为了实现上述发明目的，本发明提供以下技术方案:
一种耐热性提高的单位点β-甘露聚糖酶突变体，所述的β-甘露聚糖酶突变体为M7(D97N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第97位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQ ID NO：3所示；
或者所述的β-甘露聚糖酶突变体为M8(D107N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第107位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQ IDNO：4所示；
或者所述的β-甘露聚糖酶突变体为M9(D113N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第113位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得，氨基酸序列如SEQ IDNO：5所示；
或者所述的β-甘露聚糖酶突变体为M10(D123N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第123位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：6所示；
或者所述的β-甘露聚糖酶突变体为M11(D136N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第136位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：7所示；
或者所述的β-甘露聚糖酶突变体为M12(D151N),是由氨基酸序列为SEQID NO：1的β-甘露聚糖酶的第151位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQ IDNO：8所示；
或者所述的β-甘露聚糖酶突变体为M13(D154N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第154位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：9所示；
或者所述的β-甘露聚糖酶突变体M14(D159N),是由氨基酸序列为SEQID NO：1的β-甘露聚糖酶的第159位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQ IDNO：10所示；
或者所述的β-甘露聚糖酶突变体为M15(D197N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第197位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：11所示；
或者所述的β-甘露聚糖酶突变体为M16(D203N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第203位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：12所示；
或者所述的β-甘露聚糖酶突变体为M17(D216N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第216位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得，氨基酸序列如SEQID NO：13所示；
或者所述的β-甘露聚糖酶突变体为M18(D218N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第218位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得，氨基酸序列如SEQID NO：14所示；
或者所述的β-甘露聚糖酶突变体为M19(D225N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第225位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：15所示；
或者所述的β-甘露聚糖酶突变体为M20(D233N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第233位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：16所示；
或者所述的β-甘露聚糖酶突变体为M21(D235N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第235位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：17所示；
或者所述的β-甘露聚糖酶突变体为M22(D255N),所述的突变体M22由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第255位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQ ID NO：18所示；
或者所述的β-甘露聚糖酶突变体为M23(D258N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第258位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：19所示；
或者所述的β-甘露聚糖酶突变体为M24(D267N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第267位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：20所示；
或者所述的β-甘露聚糖酶突变体为M25(D273N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第273位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：21所示；
或者所述的β-甘露聚糖酶突变体为M26(D274N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第274位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：22所示；
或者所述的β-甘露聚糖酶突变体为M30(D341N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第341位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：23所示；
或者所述的β-甘露聚糖酶突变体为M33(D359N),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第359位氨基酸天冬氨酸(D)变为天冬酰胺(N)获得,氨基酸序列如SEQID NO：24所示；
或者所述的β-甘露聚糖酶突变体为M41(E244Q),是由氨基酸序列为SEQ ID NO：1的β-甘露聚糖酶的第244位氨基酸谷氨酸(E)变为谷氨酰胺(Q)获得,氨基酸序列如SEQ IDNO：25所示。
本发明提供了所述β-甘露聚糖酶突变体的应用
本发明提供了含有所述的β-甘露聚糖酶突变体编码基因的重组菌株，所述重组菌株为毕赤酵母X33。
本发明与现有技术相比的有益效果：
本发明提供了分别产生所述β-甘露聚糖酶突变体的氨基酸序列。基于SEQID NO：1的β-甘露聚糖酶ManAK，通过理性设计，设计筛选位点，在SEQ ID NO：1不同位点进行天冬酰胺(N)对天冬氨酸(D)以及谷氨酰胺(Q)对谷氨酸(E)的替换，合理优化蛋白质表面电荷分布，获得23株耐热性显著提高的β-甘露聚糖酶突变体。
本发明根据所取代的位置和氨基酸，与原始β-甘露聚糖酶ManAK相比，改造后的突变体于75℃条件处理，酶活力损失一半所需时间(t1/2)是原始酶ManAK的1.2～3.5倍，说明通过本发明获得的β-甘露聚糖酶糖酶突变体与野生型相比，突变体的耐热性得到显著提高，有利于实现其在商业中的应用，从而降低生产成本。
附图说明
图1为β-甘露聚糖酶突变体与原始基因ManAK耐热性比较。
具体实施方式
本发明公开了一种甘露聚糖酶突变体、其制备方法及应用、编码该甘露聚糖酶突变体的DNA分子、载体、宿主细胞,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。需要特别指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。下面结合附图和具体实施例对本发明的技术方案做进一步详细的说明，但本实施例仅用于解释本发明,而非对本发明范围的限制。
1.菌株和载体：
大肠杆菌DH5α、毕赤酵母X33、载体pPICZαA、Amp、Zeocine购自Invitrogen公司。
2.酶与试剂盒：
PCR酶及连接酶购买自Takara公司，质粒提取试剂盒及胶回收试剂盒购自Omega公司。
3.培养基配方：
大肠杆菌培养基(LB培养基)：1％胰蛋白胨、0.5％酵母提取物、1％NaCl；
LB-AMP培养基：LB培养基加入100μg/mLAMP；
YPD培养基：1％酵母提取物、2％胰蛋白胨，2％葡萄糖；
YPDZ培养基:YPD培养基中加入100μg/mLZeocine；
MD培养基：1.34％YNB，0.4mg/L生物素，2％葡萄糖；
BMGY培养基：1％酵母提取物、2％蛋白胨、100mM磷酸钾缓冲液(pH6.0)、1％甘油；
以上培养基为固体时加入2％的琼脂粉。
4.实验方法：
菌株培养条件：大肠杆菌37℃培养，酵母30℃培养。液体培养时摇床转速为200rpm。
毕赤酵母X33转化方法：
将活化的毕赤酵母X33接种到50mLYPD液体培养基中,30℃摇瓶培养至0D600为1.2～1.5后低温离心收集菌体,依次用40mL冰冷无菌水和10mL浓度为1mol/L的冰冷山梨醇溶液清洗菌体,最后用5mL山梨醇溶液重悬菌体制得酵母感受态悬液。将100uL酵母感受态与10uL线性化载体混匀后转移到预冷的电转杯中电击转化,电转化的条件为1.5kV,5msec。电击后加入1mL山梨醇溶液,转移至1.5mL离心管中30℃温育1h。5000rpm离心5min,收集菌体涂布于MD筛选平板上倒置培养,直至长出阳性单克隆。
β-甘露聚糖酶活性检测根据中华人民共和国国家标准《GB T36861-2018》进行；甘露聚糖酶活性定义是指样品在pH为5.5，温度为37℃条件下，每分钟从浓度为3mg/mL的甘露聚糖(Sigma G0753)溶液中降解释放1μmol还原糖所需要的酶量为一个酶活力单位，以U表示。
5.实施例1：β-甘露聚糖酶热稳定性位点理性设计及引物设计
基于ManAK氨基酸序列(如SEQ ID NO：1所示)，分别将ManAK中的Asp97突变为Asn97、Asp107突变为Asn107、Asp113突变为Asn113、Asp123突变为Asn123、Asp136突变为Asn136、Asp151突变为Asn151、Asp154突变为Asn154、Asp159突变为Asn159、Asp197突变为Asn197、Asp203突变为Asn203、Asp216突变为Asn216、Asp218突变为Asn218、Asp225突变为Asn225、Asp233突变为Asn233、Asp235突变为Asn235、Asp255突变为Asn255、Asp258突变为Asn258、Asp267突变为Asn267、Asp273突变为Asn273、Asp274突变为Asn274、Asp341突变为Asn341、Asp359突变为Asn359、Glu244突变为Gln244，依次获得含相应突变位点的突变体M7、M8、M9、M10、M11、M12、M13、M14、M15、M16、M17、M18、M19、M20、M21、M22、M23、M24、M25、M26、M30、M33、M41，共计23个突变体。根据ManAK核苷酸序列(如SEQ ID NO：2所示)设计PCR引物，由青岛睿博兴科有限公司合成。
6.实施例2：β-甘露聚糖酶定点突变体重组载体构建
以含ManAK的基因的重组载体pPICZαA-ManAK为模板，加入突变位点的引物，用高保真酶进行PCR扩增，产物经DpnⅠ酶切处理，热激转化DH5α大肠杆菌感受态细胞，在含Amp的LB平板上筛选阳性克隆，测序工作由青岛睿博兴科有限公司完成。
7.实施例3：β-甘露聚糖酶定点突变体表达载体构建
将实施例2中突变后测序正确的阳性克隆提取质粒并纯化，将相应质粒用SacⅠ单酶切线性化，电转入毕赤酵母X33中，MD平板筛选得到各突变体表达菌株的转化子，转接到YPD平板中活化。
8.实施例4：β-甘露聚糖酶突变体在毕赤酵母中的高通量表达验证挑取实施例3中活化后的转化子接种含1mLBMGY培养基中，于48孔板中30℃、200rmp摇床培养，24h后首次添加培养体积1％的甲醇进行酶的诱导表达，之后培养过程中每24h诱导一次。诱导表达96h后，将培养液离心获得上清，测定发酵液上清中β-甘露聚糖酶活力和热稳定性。
9.实施例5：β-甘露聚糖酶突变体在毕赤酵母中的扩大培养表达验证将实施例4中验证热稳定性提高的β-甘露聚糖酶突变体活化，接种至20mLBMGY培养基，在摇瓶中30℃、200rmp摇床培养，24h后首次添加培养体积1％的甲醇进行酶的诱导表达，之后培养过程中每24h诱导一次。诱导表达96h后，将培养液离心获得上清，测定发酵液上清中β-甘露聚糖酶活力和热稳定性。如图1所示，结果显示突变后得到的β-甘露聚糖酶M7、M8、M9、M10、M11、M12、M13、M14、M15、M16、M17、M18、M19、M20、M21、M22、M23、M24、M25、M26、M30、M33、M41在75℃条件处理，酶活力损失一半所需时间是原始酶ManAK的1.2～3.5倍,说明上述突变体耐热性菌有提高。
以上结果表明将ManAK的Asp97突变为Asn97、Asp107突变为Asn107、Asp113突变为Asn113、Asp123突变为Asn123、Asp136突变为Asn136、Asp151突变为Asn151、Asp154突变为Asn154、Asp159突变为Asn159、Asp197突变为Asn197、Asp203突变为Asn203、Asp216突变为Asn216、Asp218突变为Asn218、Asp225突变为Asn225、Asp233突变为Asn233、Asp235突变为Asn235、Asp255突变为Asn255、Asp258突变为Asn258、Asp267突变为Asn267、Asp273突变为Asn273、Asp274突变为Asn274、Asp341突变为Asn341、Asp359突变为Asn359、Glu244突变为Gln244所得到的突变体，其耐热性有进一步提高。
序列表
<110> 中国海洋大学
<120> β-甘露聚糖酶耐热突变体、重组菌及其应用
<160> 25
<170> SIPOSequenceListing 1.0
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<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
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Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
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Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
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Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
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Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
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Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
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Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
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Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
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Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
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Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
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Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
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Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
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Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
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Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
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Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
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Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
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<210> 2
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<212> DNA
<213> 白曲霉(Aspergillus candidus Link)
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actgctttgc caaaggcttc tccagctcca tcttcttctt cttcttcctc ttcttctgct 60
tctacttctt ttgcttctac ttctggtttg caatttacta ttgatggtga aactggttac 120
tttgctggta ctaactctta ctggattggt tttcttactg atgattctga tgttgatttg 180
gttatgtctc atttgaagtc ttctggtttg aagattttga gagtttgggg ttttaacgat 240
gttactactc aaccatcttc tggtactgtt tggtaccaat tgcatcaaga tggtaagtct 300
actattaaca ctggtgctga tggtttgcaa agattggatt acgttgtttc ttctgctgaa 360
caacatgata ttaagttgat tattaacttt gttaactact ggactgatta cggtggtatg 420
tctgcttacg tttctgctta cggtggttct gatgaaactg atttttacac ttctgatact 480
atgcaatctg cttaccaaac ttacattaag actgttgttg aaagatactc taactcttct 540
gctgtttttg cttgggaatt ggctaacgaa ccaagatgtc catcttgtga tacttctgtt 600
ttgtacgatt ggattgaaaa gacttctaag tttattaagg gtttggatgc tgatcatatg 660
gtttgtattg gtgatgaagg ttttggtttg aacactgatt ctgatggttc ttacccatac 720
caatttgctg aaggtttgaa ctttactaag aacttgggta ttgatactat tgattttggt 780
actttgcatt tgtacccaga ttcttggggt acttctgatg attggggtaa cggttggatt 840
tctgctcatg gtgctgcttg taaggctgct ggtaagccat gtttgttgga agaatacggt 900
gttacttcta accattgttc tgttgaatct ccatggcaaa agactgcttt gaacactact 960
ggtgtttctg ctgatttgtt ttggcaatac ggtgatgatt tgtctactgg taagtctcca 1020
gatgatggta acactattta ctacggtact tctgattacg aatgtttggt tactgatcat 1080
gttgctgcta ttggttctgc ttaa 1104
<210> 3
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
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Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
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Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
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Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
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Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
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Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asn Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
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Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
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Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
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Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
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Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
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Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
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Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
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Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
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Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
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Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
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Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
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His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
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Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
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Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
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Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
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<210> 4
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<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
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Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
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Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
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Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
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Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
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Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
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Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asn Gly Leu Gln Arg Leu
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Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
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Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
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Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
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Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
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Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
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Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
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Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
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Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
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Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
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His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 5
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 5
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
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Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
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Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
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Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
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Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
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Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
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Asn Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
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Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
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Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
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Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
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Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
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Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
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Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
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Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 6
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 6
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
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Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
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Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
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Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
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Asp Tyr Val Val Ser Ser Ala Glu Gln His Asn Ile Lys Leu Ile Ile
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Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
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Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
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Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
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Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
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Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
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Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 7
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 7
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
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Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
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Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asn Tyr Gly Gly Met Ser Ala Tyr Val
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Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
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Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
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Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
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Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
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Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
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Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
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Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 8
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 8
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asn Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 9
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 9
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
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Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
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Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asn Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 10
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 10
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asn Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 11
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 11
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asn Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 12
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 12
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asn Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 13
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 13
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asn Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 14
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 14
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asn His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 15
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 15
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asn Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 16
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 16
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asn Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 17
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 17
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asn Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 18
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 18
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asn Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 19
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 19
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asn Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 20
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 20
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asn Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 21
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 21
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asn Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 22
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 22
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asn Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 23
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 23
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asn Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 24
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 24
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Glu Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asn His Val Ala Ala Ile Gly Ser Ala
355 360 365
<210> 25
<211> 367
<212> PRT
<213> 白曲霉(Aspergillus candidus Link)
<400> 25
Thr Ala Leu Pro Lys Ala Ser Pro Ala Pro Ser Ser Ser Ser Ser Ser
1 5 10 15
Ser Ser Ser Ala Ser Thr Ser Phe Ala Ser Thr Ser Gly Leu Gln Phe
20 25 30
Thr Ile Asp Gly Glu Thr Gly Tyr Phe Ala Gly Thr Asn Ser Tyr Trp
35 40 45
Ile Gly Phe Leu Thr Asp Asp Ser Asp Val Asp Leu Val Met Ser His
50 55 60
Leu Lys Ser Ser Gly Leu Lys Ile Leu Arg Val Trp Gly Phe Asn Asp
65 70 75 80
Val Thr Thr Gln Pro Ser Ser Gly Thr Val Trp Tyr Gln Leu His Gln
85 90 95
Asp Gly Lys Ser Thr Ile Asn Thr Gly Ala Asp Gly Leu Gln Arg Leu
100 105 110
Asp Tyr Val Val Ser Ser Ala Glu Gln His Asp Ile Lys Leu Ile Ile
115 120 125
Asn Phe Val Asn Tyr Trp Thr Asp Tyr Gly Gly Met Ser Ala Tyr Val
130 135 140
Ser Ala Tyr Gly Gly Ser Asp Glu Thr Asp Phe Tyr Thr Ser Asp Thr
145 150 155 160
Met Gln Ser Ala Tyr Gln Thr Tyr Ile Lys Thr Val Val Glu Arg Tyr
165 170 175
Ser Asn Ser Ser Ala Val Phe Ala Trp Glu Leu Ala Asn Glu Pro Arg
180 185 190
Cys Pro Ser Cys Asp Thr Ser Val Leu Tyr Asp Trp Ile Glu Lys Thr
195 200 205
Ser Lys Phe Ile Lys Gly Leu Asp Ala Asp His Met Val Cys Ile Gly
210 215 220
Asp Glu Gly Phe Gly Leu Asn Thr Asp Ser Asp Gly Ser Tyr Pro Tyr
225 230 235 240
Gln Phe Ala Gln Gly Leu Asn Phe Thr Lys Asn Leu Gly Ile Asp Thr
245 250 255
Ile Asp Phe Gly Thr Leu His Leu Tyr Pro Asp Ser Trp Gly Thr Ser
260 265 270
Asp Asp Trp Gly Asn Gly Trp Ile Ser Ala His Gly Ala Ala Cys Lys
275 280 285
Ala Ala Gly Lys Pro Cys Leu Leu Glu Glu Tyr Gly Val Thr Ser Asn
290 295 300
His Cys Ser Val Glu Ser Pro Trp Gln Lys Thr Ala Leu Asn Thr Thr
305 310 315 320
Gly Val Ser Ala Asp Leu Phe Trp Gln Tyr Gly Asp Asp Leu Ser Thr
325 330 335
Gly Lys Ser Pro Asp Asp Gly Asn Thr Ile Tyr Tyr Gly Thr Ser Asp
340 345 350
Tyr Glu Cys Leu Val Thr Asp His Val Ala Ala Ile Gly Ser Ala
355 360 365

Claims (4)

1.一种耐热性提高的单位点β-甘露聚糖酶突变体，其特征在于所述β-甘露聚糖酶突变体的氨基酸序列为下列序列之一：SEQ ID NO：3、SEQ ID NO：4、SEQ ID NO：5、SEQ ID NO：6、SEQ ID NO：7、SEQ ID NO：8、SEQ ID NO：9、SEQ ID NO：10、SEQ ID NO：11、SEQ ID NO：12、SEQ ID NO：13、SEQ ID NO：14、SEQ ID NO：15、SEQ ID NO：16、SEQ ID NO：17、SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23、SEQ IDNO：24或SEQ ID NO：25。
2.含有权利要求1所述β-甘露聚糖酶突变体的编码基因的重组菌株，所述重组菌株为毕赤酵母X33。
3.权利要求1所述β-甘露聚糖酶突变体的应用。
4.权利要求2所述重组菌株的应用。
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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113201521A (zh) * 2021-06-23 2021-08-03 中国海洋大学 β-甘露聚糖酶耐热突变体M14、重组菌及其应用
CN113201520A (zh) * 2021-06-23 2021-08-03 中国海洋大学 β-甘露聚糖酶耐热突变体M15、重组菌及其应用
CN113215133A (zh) * 2021-06-23 2021-08-06 中国海洋大学 β-甘露聚糖酶耐热突变体M22、重组菌及其应用
CN113215134A (zh) * 2021-06-23 2021-08-06 中国海洋大学 β-甘露聚糖酶耐热突变体M16、重组菌及其应用
CN113322244A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M23、重组菌及其应用
CN113322247A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M19、重组菌及其应用
CN113322246A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M13、重组菌及其应用
CN113322245A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M17、重组菌及其应用
WO2023041040A1 (zh) * 2021-09-18 2023-03-23 青岛蔚蓝生物集团有限公司 耐高温甘露聚糖酶突变体
JP7467666B2 (ja) 2022-01-26 2024-04-15 シージェイ チェイルジェダン コーポレーション マンナナーゼ活性を有する変異型ポリペプチド

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102277342A (zh) * 2011-08-16 2011-12-14 青岛康地恩生物科技有限公司 甘露聚糖酶及其突变体
CN102628056A (zh) * 2012-04-24 2012-08-08 中南林业科技大学 一种耐酸耐高温β-甘露聚糖酶基因及其应用
CN102676477A (zh) * 2011-08-19 2012-09-19 济南诺能生物工程有限公司 酸性β-甘露聚糖酶基因改造及其工程菌的构建
CN102732493A (zh) * 2011-04-07 2012-10-17 中国农业大学 耐热β-甘露聚糖酶、其编码基因、重组菌和应用
CN103184206A (zh) * 2011-08-16 2013-07-03 青岛蔚蓝生物集团有限公司 甘露聚糖酶及其突变体
CN108559739A (zh) * 2018-05-11 2018-09-21 中国农业科学院饲料研究所 耐热性提高的甘露聚糖酶PMan5A突变体及其基因和应用
CN110577946A (zh) * 2018-06-07 2019-12-17 青岛红樱桃生物技术有限公司 酶活性和耐热性提高的β-甘露聚糖酶突变体及其编码基因和应用

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102732493A (zh) * 2011-04-07 2012-10-17 中国农业大学 耐热β-甘露聚糖酶、其编码基因、重组菌和应用
CN102277342A (zh) * 2011-08-16 2011-12-14 青岛康地恩生物科技有限公司 甘露聚糖酶及其突变体
CN103184206A (zh) * 2011-08-16 2013-07-03 青岛蔚蓝生物集团有限公司 甘露聚糖酶及其突变体
CN102676477A (zh) * 2011-08-19 2012-09-19 济南诺能生物工程有限公司 酸性β-甘露聚糖酶基因改造及其工程菌的构建
CN102628056A (zh) * 2012-04-24 2012-08-08 中南林业科技大学 一种耐酸耐高温β-甘露聚糖酶基因及其应用
CN108559739A (zh) * 2018-05-11 2018-09-21 中国农业科学院饲料研究所 耐热性提高的甘露聚糖酶PMan5A突变体及其基因和应用
CN110577946A (zh) * 2018-06-07 2019-12-17 青岛红樱桃生物技术有限公司 酶活性和耐热性提高的β-甘露聚糖酶突变体及其编码基因和应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
QHA94980.1: "beta-mannanase ManAK", 《GENBANK》 *
ZHEMIN LIU等: "High-level expression of a thermophilic and acidophilic β-mannanase from Aspergillus kawachii IFO 4308 with significant potential in mannooligosaccharide preparation", 《BIORESOURCE TECHNOLOGY》 *
罗长财: "一种耐高温β-甘露聚糖酶在毕赤酵母中高效表达及其耐高温机理分析", 《中国博士学位论文全文数据库 工程科技Ⅰ辑》 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113201521A (zh) * 2021-06-23 2021-08-03 中国海洋大学 β-甘露聚糖酶耐热突变体M14、重组菌及其应用
CN113201520A (zh) * 2021-06-23 2021-08-03 中国海洋大学 β-甘露聚糖酶耐热突变体M15、重组菌及其应用
CN113215133A (zh) * 2021-06-23 2021-08-06 中国海洋大学 β-甘露聚糖酶耐热突变体M22、重组菌及其应用
CN113215134A (zh) * 2021-06-23 2021-08-06 中国海洋大学 β-甘露聚糖酶耐热突变体M16、重组菌及其应用
CN113322244A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M23、重组菌及其应用
CN113322247A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M19、重组菌及其应用
CN113322246A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M13、重组菌及其应用
CN113322245A (zh) * 2021-06-23 2021-08-31 中国海洋大学 β-甘露聚糖酶耐热突变体M17、重组菌及其应用
WO2023041040A1 (zh) * 2021-09-18 2023-03-23 青岛蔚蓝生物集团有限公司 耐高温甘露聚糖酶突变体
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