CN101151106A - 疏水蛋白在处理固化矿物建材、天然石材、铸石及陶瓷表面中的用途 - Google Patents
疏水蛋白在处理固化矿物建材、天然石材、铸石及陶瓷表面中的用途 Download PDFInfo
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Abstract
本发明涉及疏水蛋白在处理固化矿物建材、天然石材、铸石或陶瓷表面中的用途，处理此类表面的方法，以及具有含疏水蛋白涂层的固化矿物建材、天然石材、铸石或陶瓷表面。
Description
发明领域
本发明涉及疏水蛋白在处理固化矿物建材、天然石材、铸石或陶瓷表面中的用途，处理此类表面的方法，以及具有含疏水蛋白涂层的固化矿物建材、天然石材、铸石或陶瓷表面。
技术背景
已知能为室内及室外的表面添加涂层，以改善其表面的持久性和/或外观。此类涂层能够例如保存、防潮、抑制污损或有助于表面清洁。
该涂层可为永久性防污涂层，例如通过荷花效应灌注的涂层，如EP-A 933 388所公开的那样。
该涂层也可为暂时性涂层。可以借助于表面清洁过程中涂布的清洁制剂中所含的物质来实现该暂时性防污效果。例如，它们可为瓷砖清洁剂。
WO 03/002620公开了二烷基氨基烷基(甲基)丙烯酸酯作为硬表面污物释放聚合物的用途，例如用于细石地板或不锈钢表面。公开的制剂含有以重量计0.1％至5％的聚合物。
DE-1 100 61 897公开了包括含硅酸盐的疏水颗粒的清洁组合物，所述颗粒能改进污物脱离并减少再次污损。所述颗粒被待清洁物质的表面摄取，进而影响其性质。
疏水蛋白为含有约100至150个氨基酸的小蛋白质，为丝状真菌例如普通裂褶菌(Schizophyllum commune)所特有。它们一般具有8个半胱氨酸单位。
疏水蛋白对界面具有显著亲和力，因此可用于表面涂层。例如，用疏水蛋白包涂铁氟龙(Teflon)，可以得到亲水表面。
疏水蛋白可分离自天然来源。但也可以通过化学方法和/或生物技术生产方法合成非天然存在的疏水蛋白。我们的在先申请DE 102005007480.4公开了生产并非天然存在的疏水蛋白的方法。
现有技术提出了疏水蛋白在多种应用中的用途。
WO 96/41882提出疏水蛋白作为乳化剂、增稠剂或表面活性剂的用途，用以赋予疏水表面亲水性、提高亲水物质的防水性、制备水包油乳液或油包水乳液。其他提示包括诸如制备膏剂或霜剂的药物应用，以及化妆品应用如皮肤保护或者生产香波或调理剂。
EP-A 1 252 516公开了于30至80℃用含疏水蛋白的溶液对窗、隐形眼镜、生物传感器、医疗设备、实施测定或贮藏用容器、船体、固体颗粒或轿车框架或车体进行涂层。
WO 03/53383公开了疏水蛋白在化妆品应用中处理角质材料的用途。
WO 03/10331公开了疏水蛋白涂层的传感器(例如测量电极)，该传感器上非共价连有其他物质如电活性物质、抗体或酶。
以上引用的参考文献无一公开过疏水蛋白处理固化矿物建材、天然石材、铸石或陶瓷表面中的用途。
发明内容
本发明的一个目的在于提供处理此类表面的新技术，由此能够获得至少一种防污和/或疏水化和/或保存效果。
我们已经发现通过使用疏水蛋白处理选自固化矿物建材、天然石材、铸石或陶瓷的材料表面，能够实现该目的。在优选实施方案中，包括疏水蛋白及至少一种溶剂的制剂用于该目的。
本发明的第二方面提供了处理表面的方法，包括将所述表面与至少一种疏水蛋白接触，其中所述表面包括选自固化矿物建材、天然石材、铸石或陶瓷的材料表面。
本发明的第三方面提供了用至少一种疏水蛋白包被的表面，所述表面包括固化矿物建材、天然石材、铸石或陶瓷。
令人惊异的是，我们发现即使极其少量的疏水蛋白也足以对固化矿物建材、石材或陶瓷的表面进行有效的防污和/或疏水性和/或保存处理。
令人惊异的是，我们发现即使极其少量的疏水蛋白也足以对硬表面进行有效的防污处理。
本发明详述如下：
根据本发明，使用至少一种疏水蛋白用于处理固化矿物建材、天然石材、铸石或陶瓷表面。也可使用多种不同疏水蛋白的混合物。
此处所用的术语“疏水蛋白”在下文是指具有通式结构(I)的多肽：
Xn-C1-X1-50-C2-X0-5-C3-X1-100-C4-X1-100-C5-X1-50-C6-X0-5-C7-X1-50-C8-Xm (I)
其中X可为20种天然氨基酸中的任一种(苯丙氨酸Phe、亮氨酸Leu、丝氨酸Ser、酪氨酸Tyr、半胱氨酸Cys、色氨酸Trp、脯氨酸Pro、组氨酸His、谷氨酰胺Gln、精氨酸Arg、异亮氨酸Ile、甲硫氨酸Met、苏氨酸Thr、天冬酰胺Asn、赖氨酸Lys、缬氨酸Val、丙氨酸Ala、天冬氨酸Asp、谷氨酸Glu、甘氨酸Gly)。各X可相同也可不同。每一例中X旁的下标为氨基酸的数目，C代表半胱氨酸、丙氨酸、丝氨酸、甘氨酸、甲硫氨酸或苏氨酸，前提是至少四个由C标识的氨基酸为半胱氨酸，下标n和m独立地为范围在0至500的自然数，优选范围为15至300。
式(I)多肽的特征还在于，相对于相同大小的水滴与非包被玻璃表面形成的接触角，该多肽的特性可使(包被后玻璃表面)水滴接触角增加至少20°、优选至少25°、更优选至少30°，且最优选至少35°，其中每一测量均在室温进行。
C1至C8代表的氨基酸优选为半胱氨酸；但它们也可被其他类似大小的氨基酸所取代，优选丙氨酸、丝氨酸、苏氨酸、甲硫氨酸或甘氨酸。然而，C1至C8中至少4个、优选至少5个、更优选至少6个、特别优选至少7个位置应当由半胱氨酸组成。本发明所用蛋白质中的半胱氨酸可以还原形式存在，或彼此之间形成二硫键。特别优选分子内形成C-C桥，尤其是其中包括至少1个、优选2个、更优选3个而最优选4个分子内二硫桥的疏水蛋白。在上述用具有相似大小的氨基酸置换半胱氨酸的情况下，最好在这样的C位置上成对置换，从而可彼此形成分子内二硫桥。
当半胱氨酸、丝氨酸、丙氨酸、甘氨酸、甲硫氨酸或苏氨酸用于X指代的位置时，通用式中单个C位置的编号可据此改变。
优选使用通用式(II)的疏水蛋白：
Xn-C1-X3-25-C2-X0-2-C3-X5-50-C4-X2-35-C5-X2-15-C6-X0-2-C7-X3-35-C8-Xm (II)
其中X、C以及X旁的下标各自定义如上，下标n和m代表从0到300的数字，且该蛋白质的特征还在于上述接触角改变。
优选使用通用式(III)的疏水蛋白：
Xn-C1-X5-9-C2-C3-X11-39-C4-X2-23-C5-X5-9-C6-C7-X6-18-C8-Xm (III)
其中X、C以及X旁的下标各自定义如上，下标n和m代表从0到200的数字，且该蛋白质的特征还在于上述接触角改变，此外C代表的氨基酸中至少6个为半胱氨酸。特别优选所有C代表的氨基酸都为半胱氨酸。
残基Xn和Xm可为与疏水蛋白天然相连的肽序列。然而，残基Xn和Xm之一或两者均可以是不与疏水蛋白天然相连的肽序列。也包括这样的Xn和/或Xm残基，其中天然存在于疏水蛋白中的肽序列由非天然存在于疏水蛋白中的肽序列所延伸。
当Xn和/或Xm为非天然存在于疏水蛋白中的肽序列时，该序列的长度一般为至少20个氨基酸，优选至少35个氨基酸，更优选至少50个氨基酸，而最优选至少100个氨基酸。此类并非天然连于疏水蛋白的残基，下文也称为融合配偶体部分。这是为了说明所述蛋白质是由疏水蛋白部分和融合配偶体部分组成的，此二者自然状态下并不一起存在。
融合配偶体部分可选自多种蛋白质。多个融合配偶体部分也可以连接于一个疏水蛋白部分，例如连于疏水蛋白部分的氨基末端(Xn)或羧基末端(Xm)。但也可以例如将两个融合配偶体部分连于本发明所用的蛋白质的一个位置上(Xn或Xm)。
特别适宜的融合配偶体部分为天然存在于微生物、特别是大肠杆菌(E.coli)或枯草芽孢杆菌(Bacillus subtilis)中的多肽。此类融合配偶体部分的实例为序列yaad(SEQ ID NO：15和16)、yaae(SEQ ID NO：17和18)及硫氧化还原蛋白。同样高度适用的是前述序列的片段或衍生物，其中仅包含部分所述序列，优选为所述序列的70％至99％，更优选80％至98％，或相对于所述序列其中个别氨基酸或核苷酸发生了改变。
还可另外修饰根据本发明所用蛋白质的多肽序列，例如通过糖基化、酰基化或化学交联(例如通过戊二醛交联)。
根据本发明使用的蛋白质的特性之一，在于改变了该蛋白质所包被表面的表面特性。通过测量用蛋白质包被表面前后的水滴接触角并确定两次测量之差，可以试验性确定表面特性的改变。
本领域技术人员原则上知道如何实施接触角测量。测量接触角的具体实验条件描述于实验部分。在其中所述的条件下，根据本发明所用的蛋白质具有增加水滴在玻璃表面接触角的特性，至少增加20°，优选至少25°，更优选至少30°。
目前已知的疏水蛋白中疏水蛋白部分的极性和非极性氨基酸位置保守，从而产生了特征性的疏水性图谱。根据生理性质和疏水性的差异，可将目前已知的疏水蛋白分为两类，I类和II类(Wessels等人，Ann.Rev.Phytopathol.，1994，32，413-437)。
I类疏水蛋白的组装膜高度不可溶(即使是升高温度在1％重量的十二烷基硫酸钠(SDS)水性溶液中也是如此)，仅通过浓缩三氟乙酸(TFA)或甲酸才可再次解离。与之相反，II类疏水蛋白的组装形式较不稳定。它们通过60％重量的乙醇或1％重量的SDS(于室温)即可再次解离。
氨基酸序列的比较揭示，II类疏水蛋白中半胱氨酸C3与半胱氨酸C4之间的区域长度明显小于I类疏水蛋白。II类疏水蛋白还比I类具有更多带电荷的氨基酸。
实施本发明尤其优选的疏水蛋白为dewA、rodA、hypA、hypB、sc3、basf1、basf2型的疏水蛋白，下文的序列表对其进行了结构表征。疏水蛋白也可仅为其部分或衍生物。也可以将多个相同或不同结构的疏水蛋白(优选2个或3个)一起连接于并非天然连接疏水蛋白的相应适宜多肽序列上。
为实施本发明特别合适的还有具有SEQ ID NO：20、22、24所示多肽序列的融合蛋白质及其编码核酸序列，特别是SEQ ID NO：19、21、23的序列。特别优选的实施方案还包括由SEQ ID NO：20、22或24所示多肽序列起始，通过取代、插入或缺失至少1个至多10个、优选5个、更优选所有氨基酸的5％而产生的蛋白质，所述蛋白质仍然具有起始蛋白质至少50％的生物特性。此处，应理解根据本发明所用蛋白质的生物特性是指上述至少20°的接触角改变。
根据本发明所用的多肽可通过肽合成的成熟技术化学制备，例如通过Merrifield固相合成。
可以利用合适的方法将天然存在的疏水蛋白从自然来源中分离出来。例如可见Wsten等人，Eur.J Cell Bio.63，122-129(1994)或WO 96/41882。
非天然融合蛋白质优选通过遗传工程方法制备，其中将编码融合配偶体的核酸序列(特别是DNA序列)与编码疏水蛋白部分的核酸序列(特别是DNA序列)组合，从而通过组合核酸序列的基因表达在宿主微生物中产生所需蛋白质。此类构建方法公开于我们在先申请DE 102005007480.4中。
用于所述构建方法的合适宿主或生产微生物包括原核细胞(包括古生菌(Archaea))或真核细胞，特别是细菌(包括嗜盐菌(halobacteria)和甲烷球菌(methanococci))、真菌、昆虫细胞、植物细胞和哺乳动物细胞，更优选大肠杆菌、枯草芽孢杆菌、巨大芽孢杆菌(Bacillus megaterium)、米曲霉(Aspergillus oryzea)、构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillusniger)、巴斯德毕赤酵母(Pichia pastoris)、假单胞菌属物种(Pseudomonasspec.)、乳酸杆菌(lactobacilli)、多形汉逊酵母(Hansenula polymorpha)、里氏木霉(Trichoderma reesei)、SF9(或相关细胞)等等。
为实施本发明，可以使用得到的调节核酸序列遗传调控下的本发明所用多肽编码核酸序列的表达构建体，以及含有至少一种这些表达构建体的载体来制备疏水蛋白。
所用表达构建体优选包括特定编码序列5’上游的启动子，以及特定编码序列3’下游的终止子序列，适用的情况下还包括各自与编码序列有效连接的其他常见调控元件。
“有效连接”是指启动子、编码序列、终止子以及适用情况下的其他调控元件的有序排列，从而使每一调控元件能够实现其表达编码序列所需的功能。
有效连接序列的实例为寻靶序列以及增强子、聚腺苷化信号等。其他调控元件包括可选择标记、扩增信号、复制起点等。合适的调控序列描述于如Goeddel，Gene Expression Technology：Methods in Enzymology 185，Academic Press，San Diego，CA(1990)中。
除了这些调控序列，在实际结构基因的上游可能还存在对这些序列的天然调控，且适用的情况下可对之进行遗传修饰，从而关闭天然调控而增强基因表达。
优选的核酸构建体最好还包括一个或多个与启动子功能性连接的上述增强子序列，以增强核酸序列的表达。其他有利的序列如其他调控元件或终止子也可插入DNA序列的3’端。
构建体中可能具有一个或多个拷贝的核酸。如为选择构建体所需，适用的情况下，所述构建体还可包括其他标记如抗生素抗性或营养缺陷型补偿基因。
例如，所述方法有利的调控序列存在于启动子如cos、tac、trp、tet、trp-tet、Ipp、lac、Ipp-lac、laclq-T7、T5、T3、gal、trc、ara、rhaP(rhaPBAD)SP6、λ-PR或imlambda-P启动子中，所述启动子最好用于革兰氏阴性细菌。其他有利的调控序列存在于例如革兰氏阳性启动子amy和SP02、酵母或真菌启动子ADC1、MFα、AC、P-60、CYC1、GAPDH、TEF、rp28、ADH中。也可使用人工启动子进行调控。
为在宿主生物中表达核酸构建体，最好将其插入载体(如质粒或噬菌体)中，从而达到基因在宿主中的最佳表达。载体以及质粒和噬菌体，还包括所有其他本身已知的载体，例如病毒(如SV40、CMV、杆状病毒和腺病毒)、转座子、IS元件、噬菌粒、粘粒以及线性或环状DNA，还有农杆菌(Agrobacterium)系统。
这些载体在宿主生物中可以自主或随染色体复制。这些载体构成了本发明的另一形式。合适质粒的实例为，大肠杆菌的pLG338、pACYC184、pBR322、pUC18、pUC19、pKC30、pRep4、pHS1、pKK223-3、pDHE19.2、pHS2、pPLc236、pMBL24、pLG200、pUR290、pIN-III”3-B1、tgt11或pBdCI，链霉菌(Streptomyces)的pIJ101、pIJ364、pIJ702或pIJ361，杆菌的pUB110、pC194或pBD214，棒状杆菌(Corynebacterium)的pSA77或pAJ667，真菌的pALS1、plL2或pBB116，酵母的2α、pAG-1、YEp6、YEp13或pEMBLYe23，或植物的pLGV23、pGHlac+、pBIN19、pAK2004或pDH51。所述质粒构成了可能质粒的一小部分。其他质粒本身已知，可见于例如书籍Cloning Vectors(Pouwels P.H.等人编辑Elsevier，Amsterdam-New York-Oxford，1985，ISBN 0 444 904018)。
为表达其他存在的基因，核酸构建体中最好还包括3’-和/或5’-末端调控序列以增强表达，所述序列根据宿主生物和基因的选择而加以选择，以达到最佳表达。
这些调控序列旨在使基因和蛋白质的表达具有特异性。例如，取决于宿主生物，这可能意味着基因仅在诱导后表达或过表达，或即刻表达和/或过表达。
优选引入基因的表达可受调控序列或因子的正面影响而增强。因而最好利用强转录信号(如启动子和/或增强子)在转录水平增强调控元件。然而，除此之外，也可以通过如提高mRNA的稳定性来增强翻译。
在另一种载体形式中，含有核酸构建体或核酸的载体也可以最好以线性DNA的形式引入微生物，并通过异源或同源重组整合于宿主生物的基因组中。线性DNA可由线性化载体如质粒组成，或仅由该核酸构建体或核酸组成。
为实现生物中异源基因的最佳表达，最好根据生物中特定的密码子使用来修饰核酸序列。通过计算机分析所研究生物的其他已知基因，可以确定密码子的使用。
将合适的启动子与合适的编码核苷酸序列及终止子或聚腺苷化信号融合，制备表达盒。常规重组和克隆技术见于例如T.Maniatis，E.F.Fritseh和J.Sambrook，Molecular Cloning：A Laboratory Manual，Cold SpringHarbor Laboratory，Cold Spring Harbor，NY(1989)，及T.J.Silhavy，M.L Berman和L.W.Enquist，Experiments with Gene Fusions，ColdSpring Harbor Laboratory，Cold Spring Harbor，NY(1984)，以及Ausubel，F.M.等人，Current Protocols in Molecular Biology，Greene PublishingAssoc.and Wiley Interscience(1987)中，可用于此目的。
为实现合适宿主生物中的表达，最好将重组核酸构建体或基因构建体插入宿主特异性的载体中，这可提供宿主中基因的最佳表达。载体本身为已知且可见例如“Cloning Vectors”(Pouwels P.H.等人编辑，Elsevier，Amsterdam-New York-Oxford，1985)。
可以利用载体制备例如由至少一个载体转化的可用于生产本发明所用多肽的重组微生物。最好将上述重组构建体引入合适的宿主系统并表达之。此处，优选使用本领域技术工人已知的成熟的克隆和转染技术以在特定表达系统中表达所述核酸，例如共沉淀、原生质体融合、电穿孔、逆转录病毒转染等。合适系统的描述见于例如Current Protocols in MolecularBiology，F.Ausubel等人编辑，Wiley Interscience，New York 1997，或Sambrook等人.Molecular Cloning：A Laboratory Manual.第2版，ColdSpring Harbor Laboratory，Cold Spring Harbor Laboratory Press，ColdSpring Harbor，NY，1989。
也可以制备同源重组的微生物。为此，可制备包括至少一部分本发明所用基因或编码序列的载体，适用的情况下，所述编码序列中引入至少一个氨基酸缺失、氨基酸添加或氨基酸取代，从而修饰(例如功能性破坏)该序列(敲除载体)。引入序列可为例如相关微生物的同源物或由哺乳动物、酵母或昆虫来源衍生而来。或者，可设计用于同源重组的载体，从而使内源性基因在同源重组时发生突变或其他改变，但仍然编码功能性蛋白质(例如改变上游调控区域由此改变内源性蛋白质的表达)。本发明所用基因的改变了的部分位于同源重组载体中。适于同源重组载体的构建见于例如Thomas，K.R.和Capecchi，M.R.(1987)Cell 51：503。
适用于本发明所用核酸或核酸构建体的重组宿主生物原则上可为任何原核或真核生物。最好使用微生物如细菌、真菌或酵母作为宿主生物。革兰氏阳性或革兰氏阴性细菌，优选肠杆菌科(Enterobacteriaceae)、假单胞菌科(Pseudomonadaceae)、根瘤菌科(Rhizobiaceae)、链霉菌科(Streptomycetaceae)或诺卡氏菌科(Nocardiaceae)的细菌，特别优选使用的细菌为埃希氏菌属(Escherichia)、假单胞菌属、链酶菌属、诺卡氏菌属、伯克霍尔德菌属(Burkholderia)、沙门氏菌属(Salmonella)、农杆菌属或红球菌属(Rhodococcus)的细菌。
根据宿主生物的不同，通过技术工人公知的方式生长或培养用于制备融合蛋白质方法的生物。微生物通常生长于液体培养基中，其中包括碳源(通常为糖的形式)、氮源(通常为有机氮源如酵母膏或盐如硫酸铵的形式)、微量元素(如铁盐、锰盐和镁盐)，以及适用情况下的维生素，温度为0℃至100℃，优选10℃至60℃，同时供以氧气。此处，营养液的pH值可保持在固定值，即培养期间可以调节也可不调节。培养可批量、半批量或连续进行。营养素可以在发酵起始时加入，或以半连续或连续的方式进行随后加料。可以通过实施例中所述的方法从生物中分离酶，也可以粗提物的形式用于反应。
同样适用于重组制备多肽或其功能生物活性片段的方法，是培养多肽生产微生物，适用的情况下诱导多肽表达，并从培养物中分离所述多肽。如果期望，也可通过该方法工业规模生产多肽。可通过已知方法培养并发酵重组微生物。例如，可以在温度20至40℃、pH6-9的TB培养基或LB培养基中繁殖细菌。合适的培养条件详细描述于T.Maniatis，E.F.Fritsch和J.Sambrook，Molecular Cloning：A Laboratory Manual，Cold SpringHarbor Laboratory，Cold Spring Harbor，NY(1989)中。
若多肽并非分泌至培养基中，则破坏细胞并通过已知蛋白质分离方法从裂解物中获取产物。可以根据需要通过高频超声、通过高压(如弗氏细胞压碎器)、通过渗透溶解、通过去污剂、裂解酶或有机溶剂的作用、通过匀浆器或通过两种或多种所列方法的组合来破坏细胞。
可以利用已知的色谱方法纯化多肽，例如分子筛色谱(凝胶过滤)，如Q琼脂糖凝胶色谱、离子交换色谱和疏水色谱，也可使用其他常用方法如超滤、结晶、盐析、透析和非变性凝胶电泳。合适的方法见述于例如Cooper,F.G.，Biochemische Arbeitsmethoden，Verlag Walter de Gruyter，Berlin，New York或Scopes，R.，Protein Purification，Springer Verlag，New York，Heidelberg，Berlin中。
通过载体系统或寡核苷酸分离重组蛋白质可能有利，所述寡核苷酸以特定核苷酸序列延伸了cDNA，从而编码改变的多肽或融合蛋白质，可用于例如简化纯化。此类适当修饰的实例有作为锚起作用的“标签”，例如称为六组氨酸锚的修饰，或能够作为抗原被抗体识别的表位(描述于如Harlow，E.和Lane，D.，1988，Antibodies：A Laboratory Manual.ColdSpring Harbor(N.Y.)Press中)。其他合适标签的实例如HA、钙调蛋白-BD、GST、MBD；几丁质-BD、链酶亲和素-BD-avi-标签、Flag-标签、T7等。这些锚可用于将蛋白质附着到固相支持物如聚合物基质上，例如，其可装载在色谱柱中，或可在微量滴定板或其他支持物上使用。相应的纯化方法可从亲和标签提供商处获得。
如所述制备的蛋白质可作为融合蛋白质直接使用，或者在切割并除去融合配偶体部分后作为“纯”疏水蛋白使用。
当需要移除融合配偶体部分时，最好在融合蛋白质的疏水蛋白部分和融合配偶体部分之间掺入潜在的裂解位点(蛋白酶的特异性识别位点)。合适的裂解位点具体包括通过生物信息学工具能够确定的那些否则将既不存在于疏水蛋白部分也不存在于融合配偶体部分中的肽序列。特别合适的有例如BrCN甲硫氨酸裂解或蛋白酶介导的Xa因子裂解、肠激酶裂解、凝血酶、TEV(烟草蚀纹病毒蛋白酶)裂解。
根据本发明待用疏水蛋白处理的表面包括选自固化矿物建材、天然石材、铸石或陶瓷的材料表面。此类表面尤其见于室内及室外的建筑领域。
本发明的固化矿物建材为基本上混合无机建材及水后由于化学和/或物理反应固化而得到的石样块状物。起始材料为在空气和水中均可固化的水硬固化建材，或仅在空气中固化的空气固化建材。建材还可以包括例如蒸汽固化建材。
此类固化建材的实例具体包括混凝土或灰泥，均可得自从水泥(如硅酸盐水泥、快干水泥或火山灰水泥)，及其与粗骨料(coarse aggregate)(如沙、砂砾或膨化材料)以及水的混合物。它们还可以已知方式包括其他无机和/或有机辅料，例如混凝土超增塑剂(superplasticizer)。其他固化建材的实例包括内部及外部应用的石膏、石灰、底灰。
天然石材包括天然存在的石材如砂岩、花岗岩、片麻岩、板岩、石灰岩或大理石。这些类型的石材不仅可用作形状不规则的碎石，也可以铸型建筑成分的形式使用，例如用作建筑石材、窗台、台阶、栏杆、门柱、铺面石板、地面石板、屋顶石板、装饰元素或雕塑。
铸石包括可与天然石材类似地应用、但并非天然来源而是一般由工业制造的建筑成分。其实例包括瓷砖、炼砖、沙石灰砖、混凝土砌块、加气混凝土砌块或膨胀粘土砌块。
术语“陶瓷”原则上为本领域技术人员公知。陶瓷为由非金属无机化合物制备物件的总称，通常可用于高温操作。
陶瓷可为粗混合物中含至少20％重量粘土材料的粘土陶瓷材料，及特种陶瓷材料，后者不合有粘土材料或仅含有低含量的粘土材料。陶瓷可为细陶瓷或粗陶瓷、多孔或致密陶瓷。陶瓷材料原则上而言可具有釉层。釉质可为彩色，也可为无色。
粘土陶瓷材料的实例包括建筑陶瓷品，例如砌体墙砖或炼砖、粘土管道、耐火砖、屋顶瓷砖、陶器产品、粘土岩产品、石灰石产品、长石产品、瓷器、硬瓷、软瓷或瓷砖，可带有釉质。
特种陶瓷品的实例包括硅石、粘土结合碳化硅、熔铸石、氧化物陶瓷绝缘材料、陶瓷滤器、碳化物陶瓷材料、电瓷、磁性陶瓷或牙科陶瓷。
其他有关陶瓷的细节可见于如Büchner等人，“IndustrielleAnorganische Chemie”，VCH Verlag，Weinheim，New York 1986，431-476页。
表面可由若干种不同材料制成。一个实例为具有由瓷砖和瓷砖灰泥组成表面的瓷砖墙面。表面还可包括不同类型的材料如金属的包埋部分。
根据本发明使用时，疏水蛋白本身就可用于处理所述表面。然而，优选疏水蛋白用作至少一种合适溶剂中的制剂或组合物。
对用于实施本发明的疏水蛋白的选择并无限制。可以使用一种疏水蛋白或多种不同的疏水蛋白。本领域技术人员会做出合适的选择。例如，可以使用融合蛋白质如yaad-Xa-dewA-his(SEQ ID NO：19)或yaad-Xa-rodA-his(SEQ ID NO：21)，其中yaad融合配偶体也可为缩短状态。
制剂用溶剂可包括水和/或有机溶剂。也可使用溶剂混合物。溶剂的种类取决于疏水蛋白、待处理表面的性质以及用途，可由本领域技术人员适当选择。
溶剂优选包括水或者水与水混溶有机溶剂的混合物。此类有机溶剂的实例包括水混溶的单羟基醇或多羟基醇，例如甲醇、乙醇、正丙醇、异丙醇、乙二醇、丙二醇或甘油。也可使用醚醇。实例包括(聚)乙二醇或(聚)丙二醇的单烷基醚，例如乙二醇单丁醚。水溶性及有机溶剂的种类和量由本领域技术人员选定。
为制备根据本发明所用的组合物，优选使用初始合成的、初始分离的和/或初始纯化的水性疏水蛋白溶液。根据其纯度，它们可能仍包括合成中的助剂残留物。但也可以通过冷冻干燥实质性地初步分离疏水蛋白，而在第二步制剂成型。
本领域技术人员可根据表面的种类和/或其规划用途，确定制剂中的疏水蛋白量。但即使相对小量也足以实现防污作用。以重量计占制剂中所有组分总量0.0001％至1％的量即可满足要求，但本发明并不仅限于该范围。该量优选为0.0005％至0.5％重量的范围，更优选为0.001％至0.1％重量的范围。
制剂还任选另外包括其他成分，例如混合材料和/或助剂。此类成分的实例具体包括表面活性剂，如阴离子、非离子、两性和/或阳离子表面活性剂。其他混合材料的实例包括酸或碱，例如羧酸或氨、缓冲体系、聚合物、无机颗粒如SiO2或硅酸盐、染料、香剂或杀虫剂。
根据本发明，将表面与疏水蛋白或包括至少一种疏水蛋白及至少一种溶剂的组合物接触，对表面进行处理。
接触可以通过例如喷洒、刷洗或滚动实施，或将整个物件浸入制剂中来实施接触。后者自然只可能用于未安装的物件。处理时间由本领域技术人员决定。可以持续几秒钟至数小时。处理后可用例如水漂洗表面，以除去过量的处理液。
处理也可以与表面清洁组合实现。通过使用包括至少一种疏水蛋白、至少一种表面活性剂及至少一种溶剂的清洁组合物实现之。
可于室温以下、室温或升高温度进行处理，例如20至100℃，优选20至60℃。
用组合物处理后，干燥处理表面。处理表面可于准室温干燥，也可升高温度干燥。
在处理及适用情况下的表面干燥之后，可升高温度例如高至120℃的温度进行表面的热后处理。热后处理也可与干燥组合进行。热后处理的温度范围优选为30至100℃，更优选范围为40至80℃，例如范围为50至70℃。处理时间由本领域技术人员决定，例如范围可为1分钟至10小时。根据处理的性质，可通过用IR辐射器辐照表面或用热蒸汽鼓吹表面实施热后处理。
本发明的方法提供了选自固化矿物建材、天然石材、铸石或陶瓷的表面，所述表面具有包括至少一种疏水蛋白的涂层。该涂层一般包括位于表面上的至少一层疏水蛋白单分子层。
本发明的处理提供了至少一种防污和/或疏水性和/或保存效果。通常情况下可以得到至少两种益处，特别是组合的疏水性和防污性。即使少量疏水蛋白也具有显著的效果。通常情况下，用仅含0.01％重量疏水蛋白的组合物处理，即可引起表面的有效改变。
可以通过原则上已知的方法确定防污效果，例如通过比较用水冲洗后未处理表面及用疏水蛋白处理后表面的污物可分离性。通过测量接触角的已知方式能够确定疏水性程度
本发明的处理对于陶瓷表面例如瓷砖尤其有效。可以同时得到防污及疏水效果。这特别在潮湿的室内如浴室中具有显著的优势。。
以下实施例对本发明进行了举例说明：
A部分：
本发明所用疏水蛋白的制备及测试
实施例1
克隆yaad-His
6
/yaaE-His
6
的初步工作
利用寡核苷酸Hal570和Hal571(Hal572/Hal573)进行聚合酶链式反应。所用的模板DNA为细菌枯草芽孢杆菌的基因组DNA。得到的PCR片段包括枯草芽孢杆菌yaaD/yaaE基因的编码序列，且其末端处分别为NcoI及BgIII的限制性切割位点。纯化PCR片段并用限制性核酸内切酶NcoI及BgIII切割之。该DNA片段用作插入物并克隆入Qiagen的pQE60载体中，所述载体事先已由限制性核酸内切酶NcoI及BgIII线性化。由此得到载体pQE60YAAD#2/pQE60YaaE#5，可用于表达分别由YAAD::HIS6和YAAE::HIS6组成的蛋白质。
Hal570：gcgcgcccatggctcaaacaggtactga
Hal571：gcagatctccagccgcgttcttgcatac
Hal572：ggccatgggattaacaataggtgtactagg
Hal573：gcagatcttacaagtgccttttgcttatattcc
实施例2
yaad疏水蛋白DewA-His
6
的克隆
利用寡核苷酸KaM 416和KaM 417进行聚合酶链式反应。所用的模板DNA为霉菌构巢曲霉的基因组DNA。得到的PCR片段包括疏水蛋白基因dewA的编码序列，且其N-末端处为Xa因子蛋白酶裂解位点。纯化PCR片段并用限制性核酸内切酶BamHI切割之。该DNA片段用作插入物并克隆入事先已由限制性核酸内切酶BamHI线性化的pQE60YAAD#2载体中。
由此得到载体#508，可用于表达由YAAD::Xa::dewA::HIS6组成的融合蛋白质。
KaM416：
GCAGCCCATCAGGGATCCCTCAGCCTTGGTACCAGCGC
KaM417：
CCCGTAGCTAGTGGATCCATTGAAGGCCGCATGAAGTTCTCCGT
CTCCGC
实施例3
yaad疏水蛋白RodA-His
6
的克隆
利用寡核苷酸KaM434和KaM435，类似于质粒#508克隆质粒#513。
KaM434：
GCTAAGCGGATCCATTGAAGGCCGCATGAAGTTCTCCATTGCTG
C
KaM435：CCAATGGGGATCCGAGGATGGAGCCAAGGG
实施例4
yaad疏水蛋白BASF1-His
6
的克隆
利用寡核苷酸KaM417和KaM418，类似于质粒#508克隆质粒#507。所用的模板DNA为人工合成的DNA序列-疏水蛋白BASF1(见附录)。
KaM417：
CCCGTAGCTAGTGGATCCATTGAAGGCCGCATGAAGTTCTCCGT
CTCCGC
KaM418：
CTGCCATTCAGGGGATCCCATATGGAGGAGGGAGACAG
实施例5
yaad疏水蛋白BASF2-His
6
的克隆
利用寡核苷酸KaM417和KaM418，类似于质粒#508克隆质粒#506。所用的模板DNA为人工合成的DNA序列-疏水蛋白BASF2(见附录)。
KaM417：
CCCGTAGCTAGTGGATCCATTGAAGGCCGCATGAAGTTCTCCGT
CTCCGC
KaM418：
CTGCCATTCAGGGGATCCCATATGGAGGAGGGAGACAG
实施例6
yaad疏水蛋白SC3-His
6
的克隆
利用寡核苷酸KaM464和KaM465，类似于质粒#508克隆质粒#526。
所用的模板DNA为普通裂褶菌cDNA(见附录)。
KaM464：CGTTAAGGATCCGAGGATGTTGATGGGGGTGC
KaM465：GCTAACAGATCTATGTTCGCCCGTCTCCCCGTCGT
实施例7
重组大肠杆菌菌株yaad疏水蛋白DewA-His
6
的发酵
于15mlGreiner试管中以表达yaad疏水蛋白DewA-His6的大肠杆菌菌株接种3mlLB液体培养基。37℃在200rpm的摇床中孵育8小时。每次用1ml预培养物接种2个含隔板和250mlLB培养基(+100μg/ml氨苄青霉素)的1IErlenmeyer烧瓶，并在37℃180rpm的摇床中孵育9小时。于20l的发酵罐中用0.5l预培养物(相对水测得OD600nm1∶10)接种13.5lLB培养基(+100μg/ml氨苄青霉素)。在OD60nm为～3.5时加入140ml100mM的IPTG。3小时后，冷却发酵罐至10℃，并通过离心除去发酵液。使用细胞沉淀进一步纯化。
实施例8
重组疏水蛋白融合蛋白质的纯化
(具有C末端His6标签的疏水蛋白融合蛋白质的纯化)
将100g细胞沉淀(100-500mg疏水蛋白)溶于50mM磷酸钠缓冲液(pH 7.5)中，至总体积为200ml并重悬之。用Ultraturrax T25型(Janke andKunkel；IKA-Labortechnik)处理悬液10分钟，然后与500单位benzonase核酸酶(Merck，Darmstadt；订货号：1.01697.0001)于室温孵育1小时，以降解核酸。在细胞破碎前，利用玻璃管(glass cartridge)(P1)进行过滤。为使细胞破碎并剪切残留的基因组DNA，于1500bar进行两次匀浆(Microfluidizer M-110EH；Microfluidics Corp.)。离心匀浆物(SorvallRC-5B，GSA Rotor，250ml离心杯，60分钟，4℃，12000rpm，23000g)，将上清液置于冰上，并将沉淀重悬于100ml磷酸钠缓冲液(pH7.5)中。离心和重悬步骤重复3次，其中第3次的磷酸钠缓冲液中含1％SDS。重悬后，将溶液搅拌1小时，然后进行最终离心(Sorvall RC-5B，GSA Rotor，250ml离心杯，60分钟，4℃，12000rpm，23000g)。根据SDS-PAGE分析，最终离心的上清液中存在疏水蛋白(图1)。该实验表明，疏水蛋白很可能以包涵体的形式存在于相应的大肠杆菌细胞中。将50ml含有疏水蛋白的上清液施加在50ml镍-琼脂糖高效17-5268-02柱(Amersham)上，其中该柱用50mM Tris-Cl缓冲液(pH8.0)平衡。用50mM Tris-Cl缓冲液(pH8.0)洗柱，随后用含有200mM咪唑的50mM Tris-Cl缓冲液(pH8.0)洗脱疏水蛋白。为除去咪唑，在50mM Tris-Cl缓冲液(pH8.0)中透析溶液。
图1显示了所制备疏水蛋白的纯化：
泳道1：施加于镍-琼脂糖柱的溶液(1∶10稀释)
泳道2：渗漏＝洗涤步骤的洗脱物
泳道3-5：OD280峰值处的洗脱级分
图1的疏水蛋白分子量为约53kD。某些较小的条带代表疏水蛋白的降解产物。
实施例9
性能测试；对疏水蛋白改变水滴在玻璃上的接触角的表征
物质：
玻璃(窗玻璃，Süddeutsche Glas，Mannheim，Germany)：
疏水蛋白浓度：100μg/ml
在50mM醋酸钠(pH4)+0.1％重量Tween20中温育玻璃片过夜(温度80℃)
然后用疏水蛋白涂层在蒸馏水中冲洗玻璃片
然后温育：10分钟/80℃/1％重量的水性十二烷基硫酸钠(SDS)蒸馏水溶液
蒸馏水洗涤
样品风干(室温)，并于室温测定5μl水滴的接触角(度数)。
在Dataphysics Contact Angle System OCA15+，软件SCA20.2.0.(2002年11月)中确定接触角测量值。根据生产商的说明进行测量。
未处理玻璃的接触角为30±5°；实施例8的功能性疏水蛋白(yaad-dewA-his6)涂层的接触角为75±5°。
B部分：
疏水蛋白在陶瓷表面防污涂层中的应用
所用溶液：
利用根据实施例8制备的融合蛋白质yaad-Xa-dewA-his(SEQ ID NO：19)的水溶液进行性能测试。疏水蛋白在溶液中的浓度为100μg/ml(0.01％重量)。
所用陶瓷表面：
陶瓷瓷砖、闪耀白，10cm×15cm(来自Novocker)，用乙醇和水擦拭。
所用污物：
利用IKW砂石土(ballast soil)实施测试(根据Seifen，Fette，le，Wachse(SFW)-Journal，124卷，14/98，1029页)。
处理方法
将2g上述浓度为100μg/ml的水性疏水蛋白溶液滴至每块瓷砖上(1.3μm疏水蛋白/cm2)，并轻轻用布使其分布于整个表面。然后瓷砖风干24小时。
随后用水冲洗瓷砖，并在装有水的玻璃杯中放置3×10分钟。每次冲洗均使用新鲜水。然后将瓷砖垂直风干。
接触角的测量和防污效果
经处理瓷砖上水滴的接触角测量值为56°(10次测量的均值)。相比之下，未处理瓷砖的接触角为20°。因此瓷砖已被显著疏水化。
通过转移移液管分别用50、100及200μgIKW砂石土弄脏经处理瓷砖以及作为对比用的未处理瓷砖，并室温干燥1小时。
然后每次用500ml水3次冲洗瓷砖。尽管这样不能将污物从未处理表面上离解下来，但在疏水蛋白预处理的瓷砖上却观察到部分污物的脱离。
因此，用疏水蛋白预处理减少了污物在陶瓷表面的附着，并导致陶瓷表面的疏水化。
序列表中DNA和多肽序列的命名
| dewA DNA及多肽序列 | SEQ ID NO：1 |
| dewA多肽序列 | SEQ ID NO：2 |
| rodA DNA及多肽序列 | SEQ ID NO：3 |
| rodA多肽序列 | SEQ ID NO：4 |
| hypA DNA及多肽序列 | SEQ ID NO：5 |
| hypA多肽序列 | SEQ ID NO：6 |
| hypB DNA及多肽序列 | SEQ ID NO：7 |
| hypB多肽序列 | SEQ ID NO：8 |
| sc3 DNA及多肽序列 | SEQ ID NO：9 |
| sc3多肽序列 | SEQ ID NO：10 |
| basf1 DNA及多肽序列 | SEQ ID NO：11 |
| basf1多肽序列 | SEQ ID NO：12 |
| basf2 DNA及多肽序列 | SEQ ID NO：13 |
| basf2 多肽序列 | SEQ ID NO：14 |
| yaad DNA及多肽序列 | SEQ ID NO：15 |
| yaad多肽序列 | SEQ ID NO：16 |
| yaae DNA及多肽序列 | SEQ ID NO：17 |
| yaae多肽序列 | SEQ ID NO：18 |
| yaad-Xa-dewA-his DNA及多肽序列 | SEQ ID NO：19 |
| yaad-Xa-dewA-his多肽序列 | SEQ ID NO：20 |
| yaad-Xa-rodA-his DNA及多肽序列 | SEQ ID NO：21 |
| yaad-Xa-rodA-his多肽序列 | SEQ ID NO：22 |
| yaad-Xa-basf1-his DNA及多肽序列 | SEQ ID NO：23 |
| yaad-Xa-basf1-his多肽序列 | SEQ ID NO：24 |
序列表
<110>巴斯福股份公司
<120>疏水蛋白在处理固化矿物建材、天然石材、铸石及陶瓷表面中的用途
<130>PF 56487
<160>24
<170>PatentIn version 3.1
<210>1
<211>405
<212>DNA
<213>basf-dewA
<220>
<221>CDS
<222>(1)..(405)
<223>
<400>1
atg cgc ttc atc gtc tct ctc ctc gcc ttc act gcc gcg gcc acc gcg 48
Met Arg Phe Ile Val Ser Leu Leu Ala Phe Thr Ala Ala Ala Thr Ala
1 5 10 15
acc gcc ctc ccg gcc tct gcc gca aag aac gcg aag ctg gcc acc tcg 96
Thr Ala Leu Pro Ala Ser Ala Ala Lys Asn Ala Lys Leu Ala Thr Ser
20 25 30
gcg gcc ttc gcc aag cag gct gaa ggc acc acc tgc aat gtc ggc tcg 144
Ala Ala Phe Ala Lys Gln Ala Glu Gly Thr Thr Cys Asn Val Gly Ser
35 40 45
atc gct tgc tgc aac tcc ccc gct gag acc aac aac gac agt ctg ttg 192
Ile Ala Cys Cys Asn Ser Pro Ala Glu Thr Asn Asn Asp Ser Leu Leu
50 55 60
agc ggt ctg ctc ggt gct ggc ctt ctc aac ggg ctc tcg ggc aac act 240
Ser Gly Leu Leu Gly Ala Gly Leu Leu Asn Gly Leu Ser Gly Asn Thr
65 70 75 80
ggc agc gcc tgc gcc aag gcg agc ttg att gac cag ctg ggt ctg ctc 288
Gly Ser Ala Cys Ala Lys Ala Ser Leu Ile Asp Gln Leu Gly Leu Leu
85 90 95
gct ctc gtc gac cac act gag gaa ggc ccc gtc tgc aag aac atc gtc 336
Ala Leu Val Asp His Thr Glu Glu Gly Pro Val Cys Lys Asn Ile Val
100 105 110
gct tgc tgc cct gag gga acc acc aac tgt gtt gcc gtc gac aac gct 384
Ala Cys Cys Pro Glu Gly Thr Thr Asn Cys Val Ala Val Asp Asn Ala
115 120 125
ggc gct ggt acc aag gct gag 405
Gly Ala Gly Thr Lys Ala Glu
130 135
<210>2
<211>135
<212>PRT
<213>basf-dewA
<400>2
Met Arg Phe Ile Val Ser Leu Leu Ala Phe Thr Ala Ala Ala Thr Ala
1 5 10 15
Thr Ala Leu Pro Ala Ser Ala Ala Lys Asn Ala Lys Leu Ala Thr Ser
20 25 30
Ala Ala Phe Ala Lys Gln Ala Glu Gly Thr Thr Cys Asn Val Gly Ser
35 40 45
Ile Ala Cys Cys Asn Ser Pro Ala Glu Thr Asn Asn Asp Ser Leu Leu
50 55 60
Ser Gly Leu Leu Gly Ala Gly Leu Leu Asn Gly Leu Ser Gly Asn Thr
65 70 75 80
Gly Ser Ala Cys Ala Lys Ala Ser Leu Ile Asp Gln Leu Gly Leu Leu
85 90 95
Ala Leu Val Asp His Thr Glu Glu Gly Pro Val Cys Lys Asn Ile Val
100 105 110
Ala Cys Cys Pro Glu Gly Thr Thr Asn Cys Val Ala Val Asp Asn Ala
115 120 125
Gly Ala Gly Thr Lys Ala Glu
130 135
<210>3
<211>471
<212>DNA
<213>basf-rodA
<220>
<221>CDS
<222>(1)..(471)
<223>
<400>3
atg aag ttc tcc att gct gcc gct gtc gtt gct ttc gcc gcc tcc gtc 48
Met Lys Phe Ser Ile Ala Ala Ala Val Val Ala Phe Ala Ala Ser Val
1 5 10 15
gcg gcc ctc cct cct gcc cat gat tcc cag ttc gct ggc aat ggt gtt 96
Ala Ala Leu Pro Pro Ala His Asp Ser Gln Phe Ala Gly Asn Gly Val
20 25 30
ggc aac aag ggc aac agc aac gtc aag ttc cct gtc ccc gaa aac gtg 144
Gly Asn Lys Gly Asn Ser Asn Val Lys Phe Pro Val Pro Glu Asn Val
35 40 45
acc gtc aag cag gcc tcc gac aag tgc ggt gac cag gcc cag ctc tct 192
Thr Val Lys Gln Ala Ser Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser
50 55 60
tgc tgc aac aag gcc acg tac gcc ggt gac acc aca acc gtt gat gag 240
Cys Cys Asn Lys Ala Thr Tyr Ala Gly Asp Thr Thr Thr Val Asp Glu
65 70 75 80
ggt ctt ctg tct ggt gcc ctc agc ggc ctc atc ggc gcc ggg tct ggt 288
Gly Leu Leu Ser Gly Ala Leu Ser Gly Leu Ile Gly Ala Gly Ser Gly
85 90 95
gcc gaa ggt ctt ggt ctc ttc gat cag tgc tcc aag ctt gat gtt gct 336
Ala Glu Gly Leu Gly Leu Phe Asp Gln Cys Ser Lys Leu Asp Val Ala
100 105 110
gtc ctc att ggc atc caa gat ctt gtc aac cag aag tgc aag caa aac 384
Val Leu Ile Gly Ile Gln Asp Leu Val Asn Gln Lys Cys Lys Gln Asn
115 120 125
att gcc tgc tgc cag aac tcc ccc tcc agc gcg gat ggc aac ctt att 432
Ile Ala Cys Cys Gln Asn Ser Pro Ser Ser Ala Asp Gly Asn Leu Ile
130 135 140
ggt gtc ggt ctc cct tgc gtt gcc ctt ggc tcc atc ctc 471
Gly Val Gly Leu Pro Cys Val Ala Leu Gly Ser Ile Leu
145 150 155
<210>4
<211>157
<212>PRT
<213>basf-rodA
<400>4
Met Lys Phe Ser Ile Ala Ala Ala Val Val Ala Phe Ala Ala Ser Val
1 5 10 15
Ala Ala Leu Pro Pro Ala His Asp Ser Gln Phe Ala Gly Asn Gly Val
20 25 30
Gly Asn Lys Gly Asn Ser Asn Val Lys Phe Pro Val Pro Glu Asn Val
35 40 45
Thr Val Lys Gln Ala Ser Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser
50 55 60
Cys Cys Asn Lys Ala Thr Tyr Ala Gly Asp Thr Thr Thr Val Asp Glu
65 70 75 80
Gly Leu Leu Ser Gly Ala Leu Ser Gly Leu Ile Gly Ala Gly Ser Gly
85 90 95
Ala Glu Gly Leu Gly Leu Phe Asp Gln Cys Ser Lys Leu Asp Val Ala
100 105 110
Val Leu Ile Gly Ile Gln Asp Leu Val Asn Gln Lys Cys Lys Gln Asn
115 120 125
Ile Ala Cys Cys Gln Asn Ser Pro Ser Ser Ala Asp Gly Asn Leu Ile
130 135 140
Gly Val Gly Leu Pro Cys Val Ala Leu Gly Ser Ile Leu
145 150 155
<210>5
<211>336
<212>DNA
<213>basf-HypA
<220>
<221>CDS
<222>(1)..(336)
<223>
<400>5
atg atc tct cgc gtc ctt gtc gct gct ctc gtc gct ctc ccc gct ctt 48
Met Ile Ser Arg Val Leu Val Ala Ala Leu Val Ala Leu Pro Ala Leu
1 5 10 15
gtt act gca act cct gct ccc gga aag cct aaa gcc agc agt cag tgc 96
Val Thr Ala Thr Pro Ala Pro Gly Lys Pro Lys Ala Ser Ser Gln Cys
20 25 30
gac gtc ggt gaa atc cat tgc tgt gac act cag cag act ccc gac cac 144
Asp Val Gly Glu Ile His Cys Cys Asp Thr Gln Gln Thr Pro Asp His
35 40 45
acc agc gcc gcc gcg tct ggt ttg ctt ggt gtt ccc atc aac ctt ggt 192
Thr Ser Ala Ala Ala Ser Gly Leu Leu Gly Val Pro Ile Asn Leu Gly
50 55 60
gct ttc ctc ggt ttc gac tgt acc ccc att tcc gtc ctt ggc gtc ggt 240
Ala Phe Leu Gly Phe Asp Cys Thr Pro Ile Ser Val Leu Gly Val Gly
65 70 75 80
ggc aac aac tgt gct gct cag cct gtc tgc tgc aca gga aat caa ttc 288
Gly Asn Asn Cys Ala Ala Gln Pro Val Cys Cys Thr Gly Asn Gln Phe
85 90 95
acc gca ttg att aac gct ctt gac tgc tct cct gtc aat gtc aac ctc 336
Thr Ala Leu Ile Asn Ala Leu Asp Cys Ser Pro Val Asn Val Asn Leu
100 105 110
<210>6
<211>112
<212>PRT
<213>basf-HypA
<400>6
Met Ile Ser Arg Val Leu Val Ala Ala Leu Val Ala Leu Pro Ala Leu
1 5 10 15
Val Thr Ala Thr Pro Ala Pro Gly Lys Pro Lys Ala Ser Ser Gln Cys
20 25 30
Asp Val Gly Glu Ile His Cys Cys Asp Thr Gln Gln Thr Pro Asp His
35 40 45
Thr Ser Ala Ala Ala Ser Gly Leu Leu Gly Val Pro Ile Asn Leu Gly
50 55 60
Ala Phe Leu Gly Phe Asp Cys Thr Pro Ile Ser Val Leu Gly Val Gly
65 70 75 80
Gly Asn Asn Cys Ala Ala Gln Pro Val Cys Cys Thr Gly Asn Gln Phe
85 90 95
Thr Ala Leu Ile Asn Ala Leu Asp Cys Ser Pro Val Asn Val Asn Leu
100 105 110
<210>7
<211>357
<212>DNA
<213>basf-HypB
<220>
<221>CDS
<222>(1)..(357)
<223>
<400>7
atg gtc agc acg ttc atc act gtc gca aag acc ctt ctc gtc gcg ctc 48
Met Val Ser Thr Phe Ile Thr Val Ala Lys Thr Leu Leu Val Ala Leu
1 5 10 15
ctc ttc gtc aat atc aat atc gtc gtt ggt act gca act acc ggc aag 96
Leu Phe Val Asn Ile Asn Ile Val Val Gly Thr Ala Thr Thr Gly Lys
20 25 30
cat tgt agc acc ggt cct atc gag tgc tgc aag cag gtc atg gat tct 144
His Cys Ser Thr Gly Pro Ile Glu Cys Cys Lys Gln Val Met Asp Ser
35 40 45
aag agc cct cag gct acg gag ctt ctt acg aag aat ggc ctt ggc ctg 192
Lys Ser Pro Gln Ala Thr Glu Leu Leu Thr Lys Asn Gly Leu Gly Leu
50 55 60
ggt gtc ctt gct ggc gtg aag ggt ctt gtt ggc gcg aat tgc agc cct 240
Gly Val Leu Ala Gly Val Lys Gly Leu Val Gly Ala Asn Cys Ser Pro
65 70 75 80
atc acg gca att ggt att ggc tcc ggc agc caa tgc tct ggc cag acc 288
Ile Thr Ala Ile Gly Ile Gly Ser Gly Ser Gln Cys Ser Gly Gln Thr
85 90 95
gtt tgc tgc cag aat aat aat ttc aac ggt gtt gtc gct att ggt tgc 336
Val Cys Cys Gln Asn Asn Asn Phe Asn Gly Val Val Ala Ile Gly Cys
100 105 110
act ccc att aat gcc aat gtg 357
Thr Pro Ile Asn Ala Asn Val
115
<210>8
<211>119
<212>PRT
<213>basf-HypB
<400>8
Met Val Ser Thr Phe Ile Thr Val Ala Lys Thr Leu Leu Val Ala Leu
1 5 10 15
Leu Phe Val Asn Ile Asn Ile Val Val Gly Thr Ala Thr Thr Gly Lys
20 25 30
His Cys Ser Thr Gly Pro Ile Glu Cys Cys Lys Gln Val Met Asp Ser
35 40 45
Lys Ser Pro Gln Ala Thr Glu Leu Leu Thr Lys Asn Gly Leu Gly Leu
50 55 60
Gly Val Leu Ala Gly Val Lys Gly Leu Val Gly Ala Asn Cys Ser Pro
65 70 75 80
Ile Thr Ala Ile Gly Ile Gly Ser Gly Ser Gln Cys Ser Gly Gln Thr
85 90 95
Val Cys Cys Gln Asn Asn Asn Phe Asn Gly Val Val Ala Ile Gly Cys
100 105 110
Thr Pro Ile Asn Ala Asn Val
115
<210>9
<211>408
<212>DNA
<213>basf-sc3
<220>
<221>CDS
<222>(1)..(408)
<223>
<400>9
atg ttc gcc cgt ctc ccc gtc gtg ttc ctc tac gcc ttc gtc gcg ttc 48
Met Phe Ala Arg Leu Pro Val Val Phe Leu Tyr Ala Phe Val Ala Phe
1 5 10 15
ggc gcc ctc gtc gct gcc ctc cca ggt ggc cac ccg ggc acg acc acg 96
Gly Ala Leu Val Ala Ala Leu Pro Gly Gly His Pro Gly Thr Thr Thr
20 25 30
ccg ccg gtt acg acg acg gtg acg gtg acc acg ccg ccc tcg acg acg 144
Pro Pro Val Thr Thr Thr Val Thr Val Thr Thr Pro Pro Ser Thr Thr
35 40 45
acc atc gcc gcc ggt ggc acg tgt act acg ggg tcg ctc tct tgc tgc 192
Thr Ile Ala Ala Gly Gly Thr Cys Thr Thr Gly Ser Leu Ser Cys Cys
50 55 60
aac cag gtt caa tcg gcg agc agc agc cct gtt acc gcc ctc ctc ggc 240
Asn Gln Val Gln Ser Ala Ser Ser Ser Pro Val Thr Ala Leu Leu Gly
65 70 75 80
ctg ctc ggc att gtc ctc agc gac ctc aac gtt ctc gtt ggc atc agc 288
Leu Leu Gly Ile Val Leu Ser Asp Leu Asn Val Leu Val Gly Ile Ser
85 90 95
tgc tct ccc ctc act gtc atc ggt gtc gga ggc agc ggc tgt tcg gcg 336
Cys Ser Pro Leu Thr Val Ile Gly Val Gly Gly Ser Gly Cys Ser Ala
100 105 110
cag acc gtc tgc tgc gaa aac acc caa ttc aac ggg ctg atc aac atc 384
Gln Thr Val Cys Cys Glu Asn Thr Gln Phe Asn Gly Leu Ile Asn Ile
115 120 125
ggt tgc acc ccc atc aac atc ctc 408
Gly Cys Thr Pro Ile Asn Ile Leu
130 135
<210>10
<211>136
<212>PRT
<213>basf-sc3
<400>10
Met Phe Ala Arg Leu Pro Val Val Phe Leu Tyr Ala Phe Val Ala Phe
1 5 10 15
Gly Ala Leu Val Ala Ala Leu Pro Gly Gly His Pro Gly Thr Thr Thr
20 25 30
Pro Pro Val Thr Thr Thr Val Thr Val Thr Thr Pro Pro Ser Thr Thr
35 40 45
Thr Ile Ala Ala Gly Gly Thr Cys Thr Thr Gly Ser Leu Ser Cys Cys
50 55 60
Asn Gln Val Gln Ser Ala Ser Ser Ser Pro Val Thr Ala Leu Leu Gly
65 70 75 80
Leu Leu Gly Ile Val Leu Ser Asp Leu Asn Val Leu Val Gly Ile Ser
85 90 95
Cys Ser Pro Leu Thr Val Ile Gly Val Gly Gly Ser Gly Cys Ser Ala
100 105 110
Gln Thr Val Cys Cys Glu Asn Thr Gln Phe Asn Gly Leu Ile Asn Ile
115 120 125
Gly Cys Thr Pro Ile Asn Ile Leu
130 135
<210>11
<211>483
<212>DNA
<213>basf-BASF1
<220>
<221>CDS
<222>(1)..(483)
<223>
<400>11
atg aag ttc tcc gtc tcc gcc gcc gtc ctc gcc ttc gcc gcc tcc gtc 48
Met Lys Phe Ser Val Ser Ala Ala Val Leu Ala Phe Ala Ala Ser Val
1 5 10 15
gcc gcc ctc cct cag cac gac tcc gcc gcc ggc aac ggc aac ggc gtc 96
Ala Ala Leu Pro Gln His Asp Ser Ala Ala Gly Asn Gly Asn Gly Val
20 25 30
ggc aac aag ttc cct gtc cct gac gac gtc acc gtc aag cag gcc acc 144
Gly Asn Lys Phe Pro Val Pro Asp Asp Val Thr Val Lys Gln Ala Thr
35 40 45
gac aag tgc ggc gac cag gcc cag ctc tcc tgc tgc aac aag gcc acc 192
Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys Ala Thr
50 55 60
tac gcc ggc gac gtc ctc acc gac atc gac gag ggc atc ctc gcc ggc 240
Tyr Ala Gly Asp Val Leu Thr Asp Ile Asp Glu Gly Ile Leu Ala Gly
65 70 75 80
ctc ctc aag aac ctc atc ggc ggc ggc tcc ggc tcc gag ggc ctc ggc 288
Leu Leu Lys Asn Leu Ile Gly Gly Gly Ser Gly Ser Glu Gly Leu Gly
85 90 95
ctc ttc gac cag tgc gtc aag ctc gac ctc cag atc tcc gtc atc ggc 336
Leu Phe Asp Gln Cys Val Lys Leu Asp Leu Gln Ile Ser Val Ile Gly
100 105 110
atc cct atc cag gac ctc ctc aac cag gtc aac aag cag tgc aag cag 384
Ile Pro Ile Gln Asp Leu Leu Asn Gln Val Asn Lys Gln Cys Lys Gln
115 120 125
aac atc gcc tgc tgc cag aac tcc cct tcc gac gcc acc ggc tcc ctc 432
Asn Ile Ala Cys Cys Gln Asn Ser Pro Ser Asp Ala Thr Gly Ser Leu
130 135 140
gtc aac ctc ggc ctc ggc aac cct tgc atc cct gtc tcc ctc ctc cat 480
Val Asn Leu Gly Leu Gly Asn Pro Cys Ile Pro Val Ser Leu Leu His
145 150 155 160
atg 483
Met
<210>12
<211>161
<212>PRT
<213>basf-BASF1
<400>12
Met Lys Phe Ser Val Ser Ala Ala Val Leu Ala Phe Ala Ala Ser Val
1 5 10 15
Ala Ala Leu Pro Gln His Asp Ser Ala Ala Gly Asn Gly Asn Gly Val
20 25 30
Gly Asn Lys Phe Pro Val Pro Asp Asp Val Thr Val Lys Gln Ala Thr
35 40 45
Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys Ala Thr
50 55 60
Tyr Ala Gly Asp Val Leu Thr Asp Ile Asp Glu Gly Ile Leu Ala Gly
65 70 75 80
Leu Leu Lys Asn Leu Ile Gly Gly Gly Ser Gly Ser Glu Gly Leu Gly
85 90 95
Leu Phe Asp Gln Cys Val Lys Leu Asp Leu Gln Ile Ser Val Ile Gly
100 105 110
Ile Pro Ile Gln Asp Leu Leu Asn Gln Val Asn Lys Gln Cys Lys Gln
115 120 125
Asn Ile Ala Cys Cys Gln Asn Ser Pro Ser Asp Ala Thr Gly Ser Leu
130 135 140
Val Asn Leu Gly Leu Gly Asn Pro Cys Ile Pro Val Ser Leu Leu His
145 150 155 160
Met
<210>13
<211>465
<212>DNA
<213>basf-BASF2
<220>
<221>CDS
<222>(1)..(465)
<223>
<400>13
atg aag ttc tcc gtc tcc gcc gcc gtc ctc gcc ttc gcc gcc tcc gtc 48
Met Lys Phe Ser Val Ser Ala Ala Val Leu Ala Phe Ala Ala Ser Val
1 5 10 15
gcc gcc ctc cct cag cac gac tcc gcc gcc ggc aac ggc aac ggc gtc 96
Ala Ala Leu Pro Gln His Asp Ser Ala Ala Gly Asn Gly Asn Gly Val
20 25 30
ggc aac aag ttc cct gtc cct gac gac gtc acc gtc aag cag gcc acc 144
Gly Asn Lys Phe Pro Val Pro Asp Asp Val Thr Val Lys Gln Ala Thr
35 40 45
gac aag tgc ggc gac cag gcc cag ctc tcc tgc tgc aac aag gcc acc 192
Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys Ala Thr
50 55 60
tac gcc ggc gac gtc acc gac atc gac gag ggc atc ctc gcc ggc ctc 240
Tyr Ala Gly Asp Val Thr Asp Ile Asp Glu Gly Ile Leu Ala Gly Leu
65 70 75 80
ctc aag aac ctc atc ggc ggc ggc tcc ggc tcc gag ggc ctc ggc ctc 288
Leu Lys Asn Leu Ile Gly Gly Gly Ser Gly Ser Glu Gly Leu Gly Leu
85 90 95
ttc gac cag tgc gtc aag ctc gac ctc cag atc tcc gtc atc ggc atc 336
Phe Asp Gln Cys Val Lys Leu Asp Leu Gln Ile Ser Val Ile Gly Ile
100 105 110
cct atc cag gac ctc ctc aac cag cag tgc aag cag aac atc gcc tgc 384
Pro Ile Gln Asp Leu Leu Asn Gln Gln Cys Lys Gln Asn Ile Ala Cys
115 120 125
tgc cag aac tcc cct tcc gac gcc acc ggc tcc ctc gtc aac ctc ggc 432
Cys Gln Asn Ser Pro Ser Asp Ala Thr Gly Ser Leu Val Asn Leu Gly
130 135 140
aac cct tgc atc cct gtc tcc ctc ctc cat atg 465
Asn Pro Cys Ile Pro Val Ser Leu Leu His Met
145 150 155
<210>14
<211>155
<212>PRT
<213>basf-BASF2
<400>14
Met Lys Phe Ser Val Ser Ala Ala Val Leu Ala Phe Ala Ala Ser Val
1 5 10 15
Ala Ala Leu Pro Gln His Asp Ser Ala Ala Gly Asn Gly Asn Gly Val
20 25 30
Gly Asn Lys Phe Pro Val Pro Asp Asp Val Thr Val Lys Gln Ala Thr
35 40 45
Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys Ala Thr
50 55 60
Tyr Ala Gly Asp Val Thr Asp Ile Asp Glu Gly Ile Leu Ala Gly Leu
65 70 75 80
Leu Lys Asn Leu Ile Gly Gly Gly Ser Gly Ser Glu Gly Leu Gly Leu
85 90 95
Phe Asp Gln Cys Val Lys Leu Asp Leu Gln Ile Ser Val Ile Gly Ile
100 105 110
Pro Ile Gln Asp Leu Leu Asn Gln Gln Cys Lys Gln Asn Ile Ala Cys
115 120 125
Cys Gln Asn Ser Pro Ser Asp Ala Thr Gly Ser Leu Val Asn Leu Gly
130 135 140
Asn Pro Cys Ile Pro Val Ser Leu Leu His Met
145 150 155
<210>15
<211>882
<212>DNA
<213>basf-yaad
<220>
<221>CDS
<222>(1)..(882)
<223>
<400>15
atg gct caa aca ggt act gaa cgt gta aaa cgc gga atg gca gaa atg 48
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
caa aaa ggc ggc gtc atc atg gac gtc atc aat gcg gaa caa gcg aaa 96
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
atc gct gaa gaa gct gga gct gtc gct gta atg gcg cta gaa cgt gtg 144
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
cca gca gat att cgc gcg gct gga gga gtt gcc cgt atg gct gac cct 192
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
aca atc gtg gaa gaa gta atg aat gca gta tct atc ccg gta atg gca 240
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
aaa gcg cgt atc gga cat att gtt gaa gcg cgt gtg ctt gaa gct atg 288
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
ggt gtt gac tat att gat gaa agt gaa gtt ctg acg ccg gct gac gaa 336
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
gaa ttt cat tta aat aaa aat gaa tac aca gtt cct ttt gtc tgt ggc 384
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
tgc cgt gat ctt ggt gaa gca aca cgc cgt att gcg gaa ggt gct tct 432
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
atg ctt cgc aca aaa ggt gag cct gga aca ggt aat att gtt gag gct 480
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
gtt cgc cat atg cgt aaa gtt aac gct caa gtg cgc aaa gta gtt gcg 528
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
atg agt gag gat gag cta atg aca gaa gcg aaa aac cta ggt gct cct 576
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
tac gag ctt ctt ctt caa att aaa aaa gac ggc aag ctt cct gtc gtt 624
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
aac ttt gcc gct ggc ggc gta gca act cca gct gat gct gct ctc atg 672
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
atg cag ctt ggt gct gac gga gta ttt gtt ggt tct ggt att ttt aaa 720
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
tca gac aac cct gct aaa ttt gcg aaa gca att gtg gaa gca aca act 768
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
cac ttt act gat tac aaa tta atc gct gag ttg tca aaa gag ctt ggt 816
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
act gca atg aaa ggg att gaa atc tca aac tta ctt cca gaa cag cgt 864
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
atg caa gaa cgc ggc tgg 882
Met Gln Glu Arg Gly Trp
290
<210>16
<211>294
<212>PRT
<213>basf-yaad
<400>16
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
Met Gln Glu Arg Gly Trp
290
<210>17
<211>591
<212>DNA
<213>basf-yaae
<220>
<221>CDS
<222>(1)..(591)
<223>
<400>17
atg gga tta aca ata ggt gta cta gga ctt caa gga gca gtt aga gag 48
Met Gly Leu Thr Ile Gly Val Leu Gly Leu Gln Gly Ala Val Arg Glu
1 5 10 15
cac atc cat gcg att gaa gca tgc ggc gcg gct ggt ctt gtc gta aaa 96
His Ile His Ala Ile Glu Ala Cys Gly Ala Ala Gly Leu Val Val Lys
20 25 30
cgt ccg gag cag ctg aac gaa gtt gac ggg ttg att ttg ccg ggc ggt 144
Arg Pro Glu Gln Leu Asn Glu Val Asp Gly Leu Ile Leu Pro Gly Gly
35 40 45
gag agc acg acg atg cgc cgt ttg atc gat acg tat caa ttc atg gag 192
Glu Ser Thr Thr Met Arg Arg Leu Ile Asp Thr Tyr Gln Phe Met Glu
50 55 60
ccg ctt cgt gaa ttc gct gct cag ggc aaa ccg atg ttt gga aca tgt 240
Pro Leu Arg Glu Phe Ala Ala Gln Gly Lys Pro Met Phe Gly Thr Cys
65 70 75 80
gcc gga tta att ata tta gca aaa gaa att gcc ggt tca gat aat cct 288
Ala Gly Leu Ile Ile Leu Ala Lys Glu Ile Ala Gly Ser Asp Asn Pro
85 90 95
cat tta ggt ctt ctg aat gtg gtt gta gaa cgt aat tca ttt ggc cgg 336
His Leu Gly Leu Leu Asn Val Val Val Glu Arg Asn Ser Phe Gly Arg
100 105 110
cag gtt gac agc ttt gaa gct gat tta aca att aaa ggc ttg gac gag 384
Gln Val Asp Ser Phe Glu Ala Asp Leu Thr Ile Lys Gly Leu Asp Glu
115 120 125
cct ttt act ggg gta ttc atc cgt gct ccg cat att tta gaa gct ggt 432
Pro Phe Thr Gly Val Phe Ile Arg Ala Pro His Ile Leu Glu Ala Gly
130 135 140
gaa aat gtt gaa gtt cta tcg gag cat aat ggt cgt att gta gcc gcg 480
Glu Asn Val Glu Val Leu Ser Glu His Asn Gly Arg Ile Val Ala Ala
145 150 155 160
aaa cag ggg caa ttc ctt ggc tgc tca ttc cat ccg gag ctg aca gaa 528
Lys Gln Gly Gln Phe Leu Gly Cys Ser Phe His Pro Glu Leu Thr Glu
165 170 175
gat cac cga gtg acg cag ctg ttt gtt gaa atg gtt gag gaa tat aag 576
Asp His Arg Val Thr Gln Leu Phe Val Glu Met Val Glu Glu Tyr Lys
180 185 190
caa aag gca ctt gta 591
Gln Lys Ala Leu Val
195
<210>18
<211>197
<212>PRT
<213>basf-yaae
<400>18
Met Gly Leu Thr Ile Gly Val Leu Gly Leu Gln Gly Ala Val Arg Glu
1 5 10 15
His Ile His Ala Ile Glu Ala Cys Gly Ala Ala Gly Leu Val Val Lys
20 25 30
Arg Pro Glu Gln Leu Asn Glu Val Asp Gly Leu Ile Leu Pro Gly Gly
35 40 45
Glu Ser Thr Thr Met Arg Arg Leu Ile Asp Thr Tyr Gln Phe Met Glu
50 55 60
Pro Leu Arg Glu Phe Ala Ala Gln Gly Lys Pro Met Phe Gly Thr Cys
65 70 75 80
Ala Gly Leu Ile Ile Leu Ala Lys Glu Ile Ala Gly Ser Asp Asn Pro
85 90 95
His Leu Gly Leu Leu Asn Val Val Val Glu Arg Asn Ser Phe Gly Arg
100 105 110
Gln Val Asp Ser Phe Glu Ala Asp Leu Thr Ile Lys Gly Leu Asp Glu
115 120 125
Pro Phe Thr Gly Val Phe Ile Arg Ala Pro His Ile Leu Glu Ala Gly
130 135 140
Glu Asn Val Glu Val Leu Ser Glu His Asn Gly Arg Ile Val Ala Ala
145 150 155 160
Lys Gln Gly Gln Phe Leu Gly Cys Ser Phe His Pro Glu Leu Thr Glu
165 170 175
Asp His Arg Val Thr Gln Leu Phe Val Glu Met Val Glu Glu Tyr Lys
180 185 190
Gln Lys Ala Leu Val
195
<210>19
<211>1329
<212>DNA
<213>basf-yaad-Xa-dewA-his
<220>
<221>CDS
<222>(1)..(1329)
<223>
<400>19
atg gct caa aca ggt act gaa cgt gta aaa cgc gga atg gca gaa atg 48
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
caa aaa ggc ggc gtc atc atg gac gtc atc aat gcg gaa caa gcg aaa 96
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
atc gct gaa gaa gct gga gct gtc gct gta atg gcg cta gaa cgt gtg 144
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
cca gca gat att cgc gcg gct gga gga gtt gcc cgt atg gct gac cct 192
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
aca atc gtg gaa gaa gta atg aat gca gta tct atc ccg gta atg gca 240
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
aaa gcg cgt atc gga cat att gtt gaa gcg cgt gtg ctt gaa gct atg 288
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
ggt gtt gac tat att gat gaa agt gaa gtt ctg acg ccg gct gac gaa 336
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
gaa ttt cat tta aat aaa aat gaa tac aca gtt cct ttt gtc tgt ggc 384
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
tgc cgt gat ctt ggt gaa gca aca cgc cgt att gcg gaa ggt gct tct 432
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
atg ctt cgc aca aaa ggt gag cct gga aca ggt aat att gtt gag gct 480
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
gtt cgc cat atg cgt aaa gtt aac gct caa gtg cgc aaa gta gtt gcg 528
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
atg agt gag gat gag cta atg aca gaa gcg aaa aac cta ggt gct cct 576
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
tac gag ctt ctt ctt caa att aaa aaa gac ggc aag ctt cct gtc gtt 624
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
aac ttt gcc gct ggc ggc gta gca act cca gct gat gct gct ctc atg 672
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
atg cag ctt ggt gct gac gga gta ttt gtt ggt tct ggt att ttt aaa 720
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
tca gac aac cct gct aaa ttt gcg aaa gca att gtg gaa gca aca act 768
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
cac ttt act gat tac aaa tta atc gct gag ttg tca aaa gag ctt ggt 816
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
act gca atg aaa ggg att gaa atc tca aac tta ctt cca gaa cag cgt 864
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
atg caa gaa cgc ggc tgg aga tcc att gaa ggc cgc atg cgc ttc atc 912
Met Gln Glu Arg Gly Trp Arg Ser Ile Glu Gly Arg Met Arg Phe Ile
290 295 300
gtc tct ctc ctc gcc ttc act gcc gcg gcc acc gcg acc gcc ctc ccg 960
Val Ser Leu Leu Ala Phe Thr Ala Ala Ala Thr Ala Thr Ala Leu Pro
305 310 315 320
gcc tct gcc gca aag aac gcg aag ctg gcc acc tcg gcg gcc ttc gcc 1008
Ala Ser Ala Ala Lys Asn Ala Lys Leu Ala Thr Ser Ala Ala Phe Ala
325 330 335
aag cag gct gaa ggc acc acc tgc aat gtc ggc tcg atc gct tgc tgc 1056
Lys Gln Ala Glu Gly Thr Thr Cys Asn Val Gly Ser Ile Ala Cys Cys
340 345 350
aac tcc ccc gct gag acc aac aac gac agt ctg ttg agc ggt ctg ctc 1104
Asn Ser Pro Ala Glu Thr Asn Asn Asp Ser Leu Leu Ser Gly Leu Leu
355 360 365
ggt gct ggc ctt ctc aac ggg ctc tcg ggc aac act ggc agc gcc tgc 1152
Gly Ala Gly Leu Leu Asn Gly Leu Ser Gly Asn Thr Gly Ser Ala Cys
370 375 380
gcc aag gcg agc ttg att gac cag ctg ggt ctg ctc gct ctc gtc gac 1200
Ala Lys Ala Ser Leu Ile Asp Gln Leu Gly Leu Leu Ala Leu Val Asp
385 390 395 400
cac act gag gaa ggc ccc gtc tgc aag aac atc gtc gct tgc tgc cct 1248
His Thr Glu Glu Gly Pro Val Cys Lys Asn Ile Val Ala Cys Cys Pro
405 410 415
gag gga acc acc aac tgt gtt gcc gtc gac aac gct ggc gct ggt acc 1296
Glu Gly Thr Thr Asn Cys Val Ala Val Asp Asn Ala Gly Ala Gly Thr
420 425 430
aag gct gag gga tct cat cac cat cac cat cac 1329
Lys Ala Glu Gly Ser His His His His His His
435 440
<210>20
<211>443
<212>PRT
<213>basf-yaad-Xa-dewA-his
<400>20
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
Met Gln Glu Arg Gly Trp Arg Ser Ile Glu Gly Arg Met Arg Phe Ile
290 295 300
Val Ser Leu Leu Ala Phe Thr Ala Ala Ala Thr Ala Thr Ala Leu Pro
305 310 315 320
Ala Ser Ala Ala Lys Asn Ala Lys Leu Ala Thr Ser Ala Ala Phe Ala
325 330 335
Lys Gln Ala Glu Gly Thr Thr Cys Asn Val Gly Ser Ile Ala Cys Cys
340 345 350
Asn Ser Pro Ala Glu Thr Asn Asn Asp Ser Leu Leu Ser Gly Leu Leu
355 360 365
Gly Ala Gly Leu Leu Asn Gly Leu Ser Gly Asn Thr Gly Ser Ala Cys
370 375 380
Ala Lys Ala Ser Leu Ile Asp Gln Leu Gly Leu Leu Ala Leu Val Asp
385 390 395 400
His Thr Glu Glu Gly Pro Val Cys Lys Asn Ile Val Ala Cys Cys Pro
405 410 415
Glu Gly Thr Thr Asn Cys Val Ala Val Asp Asn Ala Gly Ala Gly Thr
420 425 430
Lys Ala Glu Gly Ser His His His His His His
435 440
<210>21
<211>1395
<212>DNA
<213>basf-yaad-Xa-rodA-his
<220>
<221>CDS
<222>(1)..(1395)
<223>
<400>21
atg gct caa aca ggt act gaa cgt gta aaa cgc gga atg gca gaa atg 48
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
caa aaa ggc ggc gtc atc atg gac gtc atc aat gcg gaa caa gcg aaa 96
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
atc gct gaa gaa gct gga gct gtc gct gta atg gcg cta gaa cgt gtg 144
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
cca gca gat att cgc gcg gct gga gga gtt gcc cgt atg gct gac cct 192
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
aca atc gtg gaa gaa gta atg aat gca gta tct atc ccg gta atg gca 240
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
aaa gcg cgt atc gga cat att gtt gaa gcg cgt gtg ctt gaa gct atg 288
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
ggt gtt gac tat att gat gaa agt gaa gtt ctg acg ccg gct gac gaa 336
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
gaa ttt cat tta aat aaa aat gaa tac aca gtt cct ttt gtc tgt ggc 384
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
tgc cgt gat ctt ggt gaa gca aca cgc cgt att gcg gaa ggt gct tct 432
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
atg ctt cgc aca aaa ggt gag cct gga aca ggt aat att gtt gag gct 480
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
gtt cgc cat atg cgt aaa gtt aac gct caa gtg cgc aaa gta gtt gcg 528
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
atg agt gag gat gag cta atg aca gaa gcg aaa aac cta ggt gct cct 576
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
tac gag ctt ctt ctt caa att aaa aaa gac ggc aag ctt cct gtc gtt 624
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
aac ttt gcc gct ggc ggc gta gca act cca gct gat gct gct ctc atg 672
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
atg cag ctt ggt gct gac gga gta ttt gtt ggt tct ggt att ttt aaa 720
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
tca gac aac cct gct aaa ttt gcg aaa gca att gtg gaa gca aca act 768
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
cac ttt act gat tac aaa tta atc gct gag ttg tca aaa gag ctt ggt 816
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
act gca atg aaa ggg att gaa atc tca aac tta ctt cca gaa cag cgt 864
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
atg caa gaa cgc ggc tgg aga tct att gaa ggc cgc atg aag ttc tcc 912
Met Gln Glu Arg Gly Trp Arg Ser Ile Glu Gly Arg Met Lys Phe Ser
290 295 300
att gct gcc gct gtc gtt gct ttc gcc gcc tcc gtc gcg gcc ctc cct 960
Ile Ala Ala Ala Val Val Ala Phe Ala Ala Ser Val Ala Ala Leu Pro
305 310 315 320
cct gcc cat gat tcc cag ttc gct ggc aat ggt gtt ggc aac aag ggc 1008
Pro Ala His Asp Ser Gln Phe Ala Gly Asn Gly Val Gly Asn Lys Gly
325 330 335
aac agc aac gtc aag ttc cct gtc ccc gaa aac gtg acc gtc aag cag 1056
Asn Ser Asn Val Lys Phe Pro Val Pro Glu Asn Val Thr Val Lys Gln
340 345 350
gcc tcc gac aag tgc ggt gac cag gcc cag ctc tct tgc tgc aac aag 1104
Ala Ser Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys
355 360 365
gcc acg tac gcc ggt gac acc aca acc gtt gat gag ggt ctt ctg tct 1152
Ala Thr Tyr Ala Gly Asp Thr Thr Thr Val Asp Glu Gly Leu Leu Ser
370 375 380
ggt gcc ctc agc ggc ctc atc ggc gcc ggg tct ggt gcc gaa ggt ctt 1200
Gly Ala Leu Ser Gly Leu Ile Gly Ala Gly Ser Gly Ala Glu Gly Leu
385 390 395 400
ggt ctc ttc gat cag tgc tcc aag ctt gat gtt gct gtc ctc att ggc 1248
Gly Leu Phe Asp Gln Cys Ser Lys Leu Asp Val Ala Val Leu Ile Gly
405 410 415
atc caa gat ctt gtc aac cag aag tgc aag caa aac att gcc tgc tgc 1296
Ile Gln Asp Leu Val Asn Gln Lys Cys Lys Gln Asn Ile Ala Cys Cys
420 425 430
cag aac tcc ccc tcc agc gcg gat ggc aac ctt att ggt gtc ggt ctc 1344
Gln Asn Ser Pro Ser Ser Ala Asp Gly Asn Leu Ile Gly Val Gly Leu
435 440 445
cct tgc gtt gcc ctt ggc tcc atc ctc gga tct cat cac cat cac cat 1392
Pro Cys Val Ala Leu Gly Ser Ile Leu Gly Ser His His His His His
450 455 460
cac 1395
His
465
<210>22
<211>465
<212>PRT
<213>basf-yaad-Xa-rodA-his
<400>22
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
Met Gln Glu Arg Gly Trp Arg Ser Ile Glu Gly Arg Met Lys Phe Ser
290 295 300
Ile Ala Ala Ala Val Val Ala Phe Ala Ala Ser Val Ala Ala Leu Pro
305 310 315 320
Pro Ala His Asp Ser Gln Phe Ala Gly Asn Gly Val Gly Asn Lys Gly
325 330 335
Asn Ser Asn Val Lys Phe Pro Val Pro Glu Asn Val Thr Val Lys Gln
340 345 350
Ala Ser Asp Lys Cys Gly Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys
355 360 365
Ala Thr Tyr Ala Gly Asp Thr Thr Thr Val Asp Glu Gly Leu Leu Ser
370 375 380
Gly Ala Leu Ser Gly Leu Ile Gly Ala Gly Ser Gly Ala Glu Gly Leu
385 390 395 400
Gly Leu Phe Asp Gln Cys Ser Lys Leu Asp Val Ala Val Leu Ile Gly
405 410 415
Ile Gln Asp Leu Val Asn Gln Lys Cys Lys Gln Asn Ile Ala Cys Cys
420 425 430
Gln Asn Ser Pro Ser Ser Ala Asp Gly Asn Leu Ile Gly Val Gly Leu
435 440 445
Pro Cys Val Ala Leu Gly Ser Ile Leu Gly Ser His His His His His
450 455 460
His
465
<210>23
<211>1407
<212>DNA
<213>basf-yaad-Xa-BASFl-his
<220>
<221>CDS
<222>(1)..(1407)
<223>
<400>23
atg gct caa aca ggt act gaa cgt gta aaa cgc gga atg gca gaa atg 48
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
caa aaa ggc ggc gtc atc atg gac gtc atc aat gcg gaa caa gcg aaa 96
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
atc gct gaa gaa gct gga gct gtc gct gta atg gcg cta gaa cgt gtg 144
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
cca gca gat att cgc gcg gct gga gga gtt gcc cgt atg gct gac cct 192
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
aca atc gtg gaa gaa gta atg aat gca gta tct atc ccg gta atg gca 240
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
aaa gcg cgt atc gga cat att gtt gaa gcg cgt gtg ctt gaa gct atg 288
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
ggt gtt gac tat att gat gaa agt gaa gtt ctg acg ccg gct gac gaa 336
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
gaa ttt cat tta aat aaa aat gaa tac aca gtt cct ttt gtc tgt ggc 384
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
tgc cgt gat ctt ggt gaa gca aca cgc cgt att gcg gaa ggt gct tct 432
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
atg ctt cgc aca aaa ggt gag cct gga aca ggt aat att gtt gag gct 480
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
gtt cgc cat atg cgt aaa gtt aac gct caa gtg cgc aaa gta gtt gcg 528
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
atg agt gag gat gag cta atg aca gaa gcg aaa aac cta ggt gct cct 576
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
tac gag ctt ctt ctt caa att aaa aaa gac ggc aag ctt cct gtc gtt 624
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
aac ttt gcc gct ggc ggc gta gca act cca gct gat gct gct ctc atg 672
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
atg cag ctt ggt gct gac gga gta ttt gtt ggt tct ggt att ttt aaa 720
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
tca gac aac cct gct aaa ttt gcg aaa gca att gtg gaa gca aca act 768
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
cac ttt act gat tac aaa tta atc gct gag ttg tca aaa gag ctt ggt 816
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
act gca atg aaa ggg att gaa atc tca aac tta ctt cca gaa cag cgt 864
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
atg caa gaa cgc ggc tgg aga tct att gaa ggc cgc atg aag ttc tcc 912
Met Gln Glu Arg Gly Trp Arg Ser Ile Glu Gly Arg Met Lys Phe Ser
290 295 300
gtc tcc gcc gcc gtc ctc gcc ttc gcc gcc tcc gtc gcc gcc ctc cct 960
Val Ser Ala Ala Val Leu Ala Phe Ala Ala Ser Val Ala Ala Leu Pro
305 310 315 320
cag cac gac tcc gcc gcc ggc aac ggc aac ggc gtc ggc aac aag ttc 1008
Gln His Asp Ser Ala Ala Gly Asn Gly Asn Gly Val Gly Asn Lys Phe
325 330 335
cct gtc cct gac gac gtc acc gtc aag cag gcc acc gac aag tgc ggc 1056
Pro Val Pro Asp Asp Val Thr Val Lys Gln Ala Thr Asp Lys Cys Gly
340 345 350
gac cag gcc cag ctc tcc tgc tgc aac aag gcc acc tac gcc ggc gac 1104
Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys Ala Thr Tyr Ala Gly Asp
355 360 365
gtc ctc acc gac atc gac gag ggc atc ctc gcc ggc ctc ctc aag aac 1152
Val Leu Thr Asp Ile Asp Glu Gly Ile Leu Ala Gly Leu Leu Lys Asn
370 375 380
ctc atc ggc ggc ggc tcc ggc tcc gag ggc ctc ggc ctc ttc gac cag 1200
Leu Ile Gly Gly Gly Ser Gly Ser Glu Gly Leu Gly Leu Phe Asp Gln
385 390 395 400
tgc gtc aag ctc gac ctc cag atc tcc gtc atc ggc atc cct atc cag 1248
Cys Val Lys Leu Asp Leu Gln Ile Ser Val Ile Gly Ile Pro Ile Gln
405 410 415
gac ctc ctc aac cag gtc aac aag cag tgc aag cag aac atc gcc tgc 1296
Asp Leu Leu Asn Gln Val Asn Lys Gln Cys Lys Gln Asn Ile Ala Cys
420 425 430
tgc cag aac tcc cct tcc gac gcc acc ggc tcc ctc gtc aac ctc ggc 1344
Cys Gln Asn Ser Pro Ser Asp Ala Thr Gly Ser Leu Val Asn Leu Gly
435 440 445
ctc ggc aac cct tgc atc cct gtc tcc ctc ctc cat atg gga tct cat 1392
Leu Gly Asn Pro Cys Ile Pro Val Ser Leu Leu His Met Gly Ser His
450 455 460
cac cat cac cat cac 1407
His His His His His
465
<210>24
<211>469
<212>PRT
<213>basf-yaad-Xa-BASF1-his
<400>24
Met Ala Gln Thr Gly Thr Glu Arg Val Lys Arg Gly Met Ala Glu Met
1 5 10 15
Gln Lys Gly Gly Val Ile Met Asp Val Ile Asn Ala Glu Gln Ala Lys
20 25 30
Ile Ala Glu Glu Ala Gly Ala Val Ala Val Met Ala Leu Glu Arg Val
35 40 45
Pro Ala Asp Ile Arg Ala Ala Gly Gly Val Ala Arg Met Ala Asp Pro
50 55 60
Thr Ile Val Glu Glu Val Met Asn Ala Val Ser Ile Pro Val Met Ala
65 70 75 80
Lys Ala Arg Ile Gly His Ile Val Glu Ala Arg Val Leu Glu Ala Met
85 90 95
Gly Val Asp Tyr Ile Asp Glu Ser Glu Val Leu Thr Pro Ala Asp Glu
100 105 110
Glu Phe His Leu Asn Lys Asn Glu Tyr Thr Val Pro Phe Val Cys Gly
115 120 125
Cys Arg Asp Leu Gly Glu Ala Thr Arg Arg Ile Ala Glu Gly Ala Ser
130 135 140
Met Leu Arg Thr Lys Gly Glu Pro Gly Thr Gly Asn Ile Val Glu Ala
145 150 155 160
Val Arg His Met Arg Lys Val Asn Ala Gln Val Arg Lys Val Val Ala
165 170 175
Met Ser Glu Asp Glu Leu Met Thr Glu Ala Lys Asn Leu Gly Ala Pro
180 185 190
Tyr Glu Leu Leu Leu Gln Ile Lys Lys Asp Gly Lys Leu Pro Val Val
195 200 205
Asn Phe Ala Ala Gly Gly Val Ala Thr Pro Ala Asp Ala Ala Leu Met
210 215 220
Met Gln Leu Gly Ala Asp Gly Val Phe Val Gly Ser Gly Ile Phe Lys
225 230 235 240
Ser Asp Asn Pro Ala Lys Phe Ala Lys Ala Ile Val Glu Ala Thr Thr
245 250 255
His Phe Thr Asp Tyr Lys Leu Ile Ala Glu Leu Ser Lys Glu Leu Gly
260 265 270
Thr Ala Met Lys Gly Ile Glu Ile Ser Asn Leu Leu Pro Glu Gln Arg
275 280 285
Met Gln Glu Arg Gly Trp Arg Ser Ile Glu Gly Arg Met Lys Phe Ser
290 295 300
Val Ser Ala Ala Val Leu Ala Phe Ala Ala Ser Val Ala Ala Leu Pro
305 310 315 320
Gln His Asp Ser Ala Ala Gly Asn Gly Asn Gly Val Gly Asn Lys Phe
325 330 335
Pro Val Pro Asp Asp Val Thr Val Lys Gln Ala Thr Asp Lys Cys Gly
340 345 350
Asp Gln Ala Gln Leu Ser Cys Cys Asn Lys Ala Thr Tyr Ala Gly Asp
355 360 365
Val Leu Thr Asp Ile Asp Glu Gly Ile Leu Ala Gly Leu Leu Lys Asn
370 375 380
Leu Ile Gly Gly Gly Ser Gly Ser Glu Gly Leu Gly Leu Phe Asp Gln
385 390 395 400
Cys Val Lys Leu Asp Leu Gln Ile Ser Val Ile Gly Ile Pro Ile Gln
405 410 415
Asp Leu Leu Asn Gln Val Asn Lys Gln Cys Lys Gln Asn Ile Ala Cys
420 425 430
Cys Gln Asn Ser Pro Ser Asp Ala Thr Gly Ser Leu Val Asn Leu Gly
435 440 445
Leu Gly Asn Pro Cys Ile Pro Val Ser Leu Leu His Met Gly Ser His
450 455 460
His His His His His
465
Claims (9)
1.疏水蛋白在处理选自固化矿物建材、天然石材、铸石或陶瓷的材料表面中的用途。
2.权利要求1的用途，其中通过使用包括溶剂及至少一种疏水蛋白的组合物实现所述处理。
3.处理表面的方法，包括将所述表面与至少一种疏水蛋白接触，其中所述表面包括选自固化矿物建材、天然石材、铸石或陶瓷的材料表面。
4.权利要求3的方法，其中通过使用包括溶剂及至少一种疏水蛋白的组合物实现所述处理。
5.权利要求3或4的方法，其中所述溶剂包括水。
6.权利要求3至5中任一项的方法，其中所述组合物中疏水蛋白的量以重量计占所述制剂所有组分总量0.0001％至1％的范围。
7.由至少一种疏水蛋白涂层的表面，所述表面包括选自固化矿物建材、天然石材、铸石或陶瓷的材料表面。
8.权利要求7的涂层表面，其特征在于防污。
9.权利要求7或8的涂层表面，其特征在于疏水性。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE102005014843.3 | 2005-03-30 | ||
| DE102005014843A DE102005014843A1 (de) | 2005-03-30 | 2005-03-30 | Verwendung von Hydrophobinen zur Oberflächenbehandlung von gehärteten mineralischen Baustoffen, Naturstein, Kunststein und Keramiken |
| DE102005036339.3 | 2005-07-29 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101151106A true CN101151106A (zh) | 2008-03-26 |
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ID=37055188
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006800102386A Pending CN101151106A (zh) | 2005-03-30 | 2006-03-28 | 疏水蛋白在处理固化矿物建材、天然石材、铸石及陶瓷表面中的用途 |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN101151106A (zh) |
| DE (1) | DE102005014843A1 (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110325222A (zh) * | 2017-01-25 | 2019-10-11 | 生活技术有限公司 | 使用良性细菌清洁内部空间和技术设备中的表面的方法 |
| CN114800746A (zh) * | 2022-05-13 | 2022-07-29 | 淮阴师范学院 | 利用杂色云芝制备纤维板的方法 |
-
2005
- 2005-03-30 DE DE102005014843A patent/DE102005014843A1/de not_active Withdrawn
-
2006
- 2006-03-28 CN CNA2006800102386A patent/CN101151106A/zh active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110325222A (zh) * | 2017-01-25 | 2019-10-11 | 生活技术有限公司 | 使用良性细菌清洁内部空间和技术设备中的表面的方法 |
| CN114800746A (zh) * | 2022-05-13 | 2022-07-29 | 淮阴师范学院 | 利用杂色云芝制备纤维板的方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| DE102005014843A1 (de) | 2006-10-19 |
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