Protein engineering of glucoamylase to increase ph optimum,...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/56 (2006.01) C12N 9/34 (2006.01) C12Q 1/40 (2006.01)

Patent

CA 2259958

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 formation for cumulative improvements in the engineered glucoamylases.

Cette invention concerne une glucoamylase fongique qui comprend une paire de mutation Asn20Cys couplé à Ala27Cys, et dans laquelle une liaison disulfure est formée entre les deux éléments de la paire. Cette mutation permet d'accroître la stabilité thermique et de réduire la formation d'isomaltose en ce qui concerne l'enzyme. Cette invention concerne également une glucoamylase fongique contenant une mutation 311-314Loop, et dans laquelle la formation d'isomaltose est également réduite grâce à la mutation. Cette invention concerne en outre une glucoamylase fongique contenant un Ser411Ala de mutation, et dans laquelle il est possible d'obtenir un pH optimal et de réduire la formation d'isomaltose grâce à la mutation. Cette invention concerne enfin des combinaisons des mutations dans les glucoamylases obtenues par génie génétique, ainsi que des combinaisons avec d'autres mutations de glucoamylase qui permettent d'accroître la stabilité thermique, d'obtenir un pH optimal, et de réduire la formation d'isomaltose en vue d'améliorations cumulées dans les glucoamylases ainsi obtenues.

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