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    Production and secretion of proteins of bacterial origin in filamentous fungi
    4.
    发明授权
    Production and secretion of proteins of bacterial origin in filamentous fungi 有权
    细菌起源于丝状真菌的蛋白质的生产和分泌

    公开(公告)号：US07816129B2

    公开(公告)日：2010-10-19

    申请号：US10286993

    申请日：2002-08-13

    申请人： Arja Mäntylä ,  Marja Paloheimo ,  Raija Lantto ,  Richard Fagerström ,  Tarja Lahtinen ,  Pirkko Suominen ,  Jari Vehmaanperä

    发明人： Arja Mäntylä ,  Marja Paloheimo ,  Raija Lantto ,  Richard Fagerström ,  Tarja Lahtinen ,  Pirkko Suominen ,  Jari Vehmaanperä

    IPC分类号： C12N15/00 ,  C12N9/00 ,  C12N9/24 ,  C12N9/30 ,  C07H21/04

    CPC分类号： C12Y302/01032 ,  C12N9/2434 ,  C12N9/248 ,  C12N15/80 ,  C12Y302/01001 ,  C12Y302/01003 ,  C12Y302/01004 ,  C12Y302/01006 ,  C12Y302/01008 ,  C12Y302/01025 ,  C12Y302/01039 ,  C12Y302/01091 ,  D21C5/005

    摘要： The present invention is related to an improved production of bacterial proteins in filamentous fungus, e.g. in Trichoderma and Aspergillus. The improvement is achieved by constructing expression vectors, which comprise the bacterial protein encoding DNA sequences fused in frame with a DNA sequence encoding a filamentous fungus secretable protein or one or more functional domains of said protein. Filamentous fungus hosts transformed with such expression vectors secrete the desired proteins or enzymes, especially xylanases or cellulases originating from bacteria or more preferably from actinornycetes into the culture medium of the host. The desired proteins or enzymes can be used directly from the culture medium after separation of host cells or recovered and treated using down-stream processes, which are appropriate for the respective application. Xylanases or cellulases from actinomycetes produced using the above expression vectors are most suitable for treating plant derived materials e.g. in pulp and paper industries.

    摘要翻译： 本发明涉及丝状真菌中细菌蛋白质的改进生产，例如， 在木霉属和曲霉属中。 通过构建表达载体来实现改善，所述表达载体包含编码与编码丝状真菌可分泌蛋白质的DNA序列或所述蛋白质的一个或多个功能结构域框架融合的DNA序列的细菌蛋白质。 用这种表达载体转化的丝状真菌宿主分泌所需的蛋白质或酶，特别是源自细菌的木聚糖酶或纤维素酶，或更优选从放线菌素分泌到宿主的培养基中。 所需的蛋白质或酶可以在分离宿主细胞之后直接从培养基中使用，或者使用适合于各自应用的下游过程进行回收和处理。 使用上述表达载体产生的放线菌的木聚糖酶或纤维素酶最适合于处理植物来源的材料，例如 在纸浆和造纸工业。

    Method and DNA constructs for increasing the production level of carbohydrate degrading enzymes in filamentous fungi
    9.
    发明授权
    Method and DNA constructs for increasing the production level of carbohydrate degrading enzymes in filamentous fungi 有权
    用于提高丝状真菌中碳水化合物降解酶生产水平的方法和DNA构建体

    公开(公告)号：US07348172B2

    公开(公告)日：2008-03-25

    申请号：US11108163

    申请日：2005-04-18

    申请人： Marja Paloheimo ,  Arja Mantyla ,  Sanna Leskinen ,  Richard Fagerstrom ,  Jarno Kallio ,  Terhi Puranen ,  Raija Lantto ,  Pirkko Suominen

    发明人： Marja Paloheimo ,  Arja Mantyla ,  Sanna Leskinen ,  Richard Fagerstrom ,  Jarno Kallio ,  Terhi Puranen ,  Raija Lantto ,  Pirkko Suominen

    IPC分类号： C12P21/06 ,  C12N9/88 ,  C12N1/15 ,  C12N15/70 ,  C12N15/74 ,  C12Q1/68 ,  C07H21/04

    CPC分类号： C12N9/248 ,  C12N15/80 ,  C12Y302/01008

    摘要： The invention is related to a method and DNA constructs for obtaining in a filamentous fungus host a higher production level of a carbohydrate degrading (CD) enzyme, having in its original state a catalytic module (CAT) and a carbohydrate binding module (CBM) separated by a linker region. The DNA construct comprising a truncated actinomycetes, preferably Nonomuraea flexuosa (NJ) derived DNA sequence encoding a truncated form of the CD enzyme, for example Nf Xyn11A, Nf Xyn10A, and is introduced into a filamentous fungal host. Said CD enzyme contains the catalytically active region of CAT but lacks part or all of the CBM, or all of the CBM and part or all of the linker region and is expressed and secreted under the control of regulatory sequences comprising at least a signal sequence, but also promoters, terminators and DNA sequences encoding a secretable carrier protein or domains thereof, preferably originating from filamentous fungi are included. The production level obtained with DNA sequence having the shortened DNA sequence encoding the truncated form of the CD enzyme is higher than the production level obtained with DNA construct encoding the corresponding full length CD enzyme.

    摘要翻译： 本发明涉及一种用于在丝状真菌宿主中获得更高生产水平的降解碳水化合物（CD）酶的方法和DNA构建体，其具有在其初始状态下分离的催化模块（CAT）和碳水化合物结合模块（CBM） 通过接头区域。 该DNA构建体包含截短的放线菌，优选非曲霉（NJ）衍生的编码CD酶的截短形式的DNA序列，例如Nf Xyn11A，Nf Xyn10A，并被引入丝状真菌宿主中。 所述CD酶含有CAT的催化活性区，但缺少部分或全部CBM，或全部CBM和部分或全部接头区，并且在包含至少一个信号序列的调控序列的控制下表达和分泌， 而且还包括编码可分泌载体蛋白或其结构域的启动子，终止子和DNA序列，优选来源于丝状真菌。 用缩短的编码CD酶的截短形式的DNA序列的DNA序列获得的生产水平高于用编码相应的全长CD酶的DNA构建体获得的产生水平。