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2. 发明授权

US06537792B1 Protein engineering of glucoamylase to increase pH optimum, substrate specificity and thermostability 失效
标题翻译：葡糖淀粉酶的蛋白质工程，以提高pH最佳，底物特异性和热稳定性
公开(公告)号：US06537792B1
公开(公告)日：2003-03-25
申请号：US09236063
申请日：1999-01-22
申请人： Martin J. Allen , Tsuei-Yun Fang , Yuxing Li , Hsuan-Liang Liu , Hsiu-Mei Chen , Pedro Coutinho , Richard Honzatko , Clark Ford
发明人： Martin J. Allen , Tsuei-Yun Fang , Yuxing Li , Hsuan-Liang Liu , Hsiu-Mei Chen , Pedro Coutinho , Richard Honzatko , Clark Ford
IPC分类号： C12N924
CPC分类号： C12N9/2428
摘要： A fungal glucoamylase including a mutation pair Asn20Cys coupled with Ala27Cys forming a disulfide bond between the two members of the pair. The mutation provides increased thermal stability and reduced isomaltose formation to the enzyme. A fungal glucoamylase including a 311-314Loop mutation wherein reduced isomaltose formation is provided by the mutation is also provided. A fungal glucoamylase including a mutation Ser411Ala wherein increased pH optimum and reduced isomaltose formation is provided by the mutation is also provided. Combinations of the mutations in engineered glucoamylases are also provided as are combinations with other glucoamylase mutations that provide increased thermal stability, increased pH optimum and reduced isomaltose froamtion for cumulative improvements in the engineered glucoamylases. Also provided is a fungal glucoamylase including a mutation of Ser30Pro coupled with at least two stabilizing mutations forming a disulfide bond between the two stabilizing members. A fungal glucoamylase including a Ser30Pro/Gly137Ala/311-314 Loop is provided. A fungal glucoamylase including a mutation Ser30Pro/Glu137Ala/Ser411Ala is also provided.
摘要翻译：一种真菌葡糖淀粉酶，其包括与成对的两个成员之间形成二硫键的Ala27Cys偶联的突变对Asn20Cys。该突变提供了增加的热稳定性和减少异黄酮对酶的形成。还提供了包含311-314Loop突变的真菌葡糖淀粉酶，其中通过突变提供了减少的异麦芽糖形成。还提供了包括突变Ser411Ala的真菌葡糖淀粉酶，其中通过突变提供了增加的pH最佳和降低的异麦芽糖形成。还提供了工程化葡糖淀粉酶突变的组合，与其它葡糖淀粉酶突变的组合一起提供，其提供增加的热稳定性，增加的pH最佳值和降低的异麦芽糖减少，用于工程化葡糖淀粉酶的累积改进。还提供了真菌葡糖淀粉酶，其包括Ser30Pro突变与至少两个在两个稳定化合物之间形成二硫键的稳定突变。提供了包含Ser30Pro / Gly137Ala / 311-314环的真菌葡糖淀粉酶。还提供了包含突变Ser30Pro / Glu137Ala / Ser411Ala的真菌葡糖淀粉酶。

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