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    • 6. 发明授权
    • Method and DNA constructs for increasing the production level of carbohydrate degrading enzymes in filamentous fungi
    • 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
    • US07348172B2
    • 2008-03-25
    • US11108163
    • 2005-04-18
    • Marja PaloheimoArja MantylaSanna LeskinenRichard FagerstromJarno KallioTerhi PuranenRaija LanttoPirkko Suominen
    • Marja PaloheimoArja MantylaSanna LeskinenRichard FagerstromJarno KallioTerhi PuranenRaija LanttoPirkko Suominen
    • C12P21/06C12N9/88C12N1/15C12N15/70C12N15/74C12Q1/68C07H21/04
    • C12N9/248C12N15/80C12Y302/01008
    • 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.
    • 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。
    • 8. 发明申请
    • Method and DNA constructs for increasing the production level of carbohydrate degrading enzymes in filamentous fungi
    • 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
    • US20060014247A1
    • 2006-01-19
    • US11108163
    • 2005-04-18
    • Marja PaloheimoArja MantylaSanna LeskinenRichard FagerstromJarno KallioTerhi PuranenRaija LanttoPirkko Suominen
    • Marja PaloheimoArja MantylaSanna LeskinenRichard FagerstromJarno KallioTerhi PuranenRaija LanttoPirkko Suominen
    • C12N9/88C12N1/16C12N15/74C07H21/04C12P21/06
    • C12N9/248C12N15/80C12Y302/01008
    • 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.
    • 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。