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PROCEDURES AND DNA CONSTRUCTIONS FOR INCREASING THE PRODUCTION LEVEL OF ENZYMES BASED FROM COAL HYDRATE IN FILAMENTS

机译:用于增加长丝中水合物的酶生产水平的方法和DNA的构建

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 (Nf) derived DNA sequence encoding a truncated form of the CD enzyme, for example Nf Xyn1 1A, 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.
机译:本发明涉及用于在丝状真菌宿主中获得较高生产水平的碳水化合物降解(CD)酶的方法和DNA构建体,所述碳水化合物降解酶在其原始状态下具有分离的催化模块(CAT)和碳水化合物结合模块(CBM)。通过链接区。将包含截短的放线菌,优选地Nonomuraea flexuosa(Nf)衍生的DNA序列的DNA构建体编码CD酶的截短形式,例如Nf Xyn1 1A,Nf Xyn10A,并将其引入丝状真菌宿主中。所述CD酶包含CAT的催化活性区域,但缺少部分或全部CBM,或全部CBM和部分或全部接头区域,并在包含至少一个信号序列的调节序列的控制下表达和分泌,但是还包括优选地来源于丝状真菌的启动子,终止子和编码可分泌的载体蛋白或其结构域的DNA序列。用具有缩短的编码CD酶截短形式的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构建体获得的生产水平。

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