Maltooligosyl trehalose trehalohydrolase (MTHase) mutant and application thereof
AssigneeJiangnan University
InventorsJing Wu, Lingqia Su, Zhendong Wang
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Abstract
The present invention discloses a maltooligosyl trehalose trehalohydrolase (MTHase) mutant and application thereof, belonging to the technical fields of gene engineering and enzyme engineering. The present invention provides a series of MTHase mutants to prepare trehalose, having a better effect. Further, the MTHase mutant is expressed in Escherichia coli BL21 (DE3) and the enzyme is optimized by fermentation, which can significantly increase the yield of enzymes.
Claims7
A mutant of an isolated maltooligosyl trehalose trehalohydrolase, consisting of mutations compared with the amino acid sequence of a parent maltooligosyl trehalose trehalohydrolase obtained from Sulfolobus acidocaldarius as set forth in SEQ ID NO.2, wherein the mutations are selected from the group consisting of:
- mutation of methionine (Met) at position 140 to leucine (Leu), and the mutant is named M140L;
- mutation of leucine (Leu) at position 202 to proline (Pro), and the mutant is named L202P;
- mutation of leucine (Leu) at position 218 to aspartic acid (Asp), and the mutant is named L218D;
- mutation of phenylalanine (Phe) at position 338 to serine (Ser), and the mutant is named F338S;
- mutation of isoleucine (Ile) at position 404 to threonine (Thr), and the mutant is named 1404T;
- mutation of leucine (Leu) at position 202 to proline (Pro), and mutation of tyrosine (Tyr) at position 323 to glycine (Gly), and the mutant is named L202P/Y323G;
- mutation of leucine (Leu) at position 202 to proline (Pro), mutation of leucine (Leu) at position 218 to aspartic acid (Asp), and mutation of tyrosine (Tyr) at position 323 to glycine (Gly), and the mutant is named L202P/L218D/Y323G;
- mutation of methionine (Met) at position 140 to leucine (Leu), mutation of leucine (Leu) at position 202 to proline (Pro), mutation of leucine (Leu) at position 218 to aspartic acid (Asp), and mutation of tyrosine (Tyr) at position 323 to glycine (Gly), and the mutant is named M140L/L202P/L218D/Y323G;
- mutation of methionine (Met) at position 140 to leucine (Leu), mutation of leucine (Leu) at position 202 to proline (Pro), mutation of leucine (Leu) at position 218 to aspartic acid (Asp), mutation of tyrosine (Tyr) at position 323 to glycine (Gly), and mutation of phenylalanine (Phe) at position 338 to serine (Ser), and the mutant is named M140L/L202P/L218D/Y323G/F338S;
- and
- mutation of methionine (Met) at position 140 to leucine (Leu), mutation of leucine (Leu) at position 202 to proline (Pro), mutation of leucine (Leu) at position 218 to aspartic acid (Asp), mutation of tyrosine (Tyr) at position 323 to glycine (Gly), mutation of phenylalanine (Phe) at position 338 to serine (Ser), and mutation of isoleucine (Ile) at position 404 to threonine (Thr), and the mutant is named M140L/L202P/L218D/Y323G/F338S/I404T;
- and wherein the mutant has increased enzymatic activity of maltooligosyl trehalose trehalohydrolase compared to the parent maltooligosyl trehalose trehalohydrolase.
Dependencies are inferred from the claim text.