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Genetically engineered bacteria highly expressing α-glucosidase and its construction method

A technology of glucosidase and genetically engineered bacteria, applied in the field of genetic engineering, can solve the problems of undiscovered patent publications and the like, and achieve the effects of reducing the cost of extraction and purification, the transformation method is simple, and the production cost is reduced

Inactive Publication Date: 2016-01-20
TIANJIN UNIV OF SCI & TECH
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Problems solved by technology

[0005] Through the search, no patent publications related to the patent application of the present invention have been found

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  • Genetically engineered bacteria highly expressing α-glucosidase and its construction method
  • Genetically engineered bacteria highly expressing α-glucosidase and its construction method
  • Genetically engineered bacteria highly expressing α-glucosidase and its construction method

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[0033] Below in conjunction with the examples, the present invention is further described, the following examples are illustrative, not limiting, and the protection scope of the present invention cannot be limited by the following examples.

[0034] The main idea of ​​the present invention is: the α-glucosidase gene is cloned and placed in other host strains for heterologous expression, because of the relationship of glycosylation, the properties of the transglycosidase will change, affecting the conversion rate of IMO, and at the same time There is also the problem of codon preference in source expression. Only by introducing it into its own host strain and performing homologous expression in the original strain can its yield be increased without changing its properties. Aspergillus niger is a strain producing α-glucosidase. It is not only an excellent expression system, but also a food-grade safe production strain with strong protein secretion ability. Therefore, the construct...

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Abstract

The invention relates to a genetically engineered strain efficiently expressing alpha-transglucosidase. A host bacterium of the genetically engineered strain is aspergillus niger, and the genetically engineered strain comprises an alpha-transglucosidase gene expression cassette. A genetically engineered bacterium can efficiently express alpha-transglucosidase, and improves the secretion efficiency of alpha-transglucosidase. In addition, alpha-transglucosidase is secreted to the outside of a cell, so that the cost of alpha-transglucosidase extraction purification is lowered, industrial materials are saved, the production cost is lowered, and the genetically engineered strain has an obvious social benefit. The enzyme activity of a crude enzyme obtained after the strain is fermented is improved by 4.0-12.1 times in comparison to a starting strain; a foundation is laid for an application of alpha-transglucosidase in the food industry and industrial production; a bottleneck that alpha-transglucosidase depends on import is broken; and the genetically engineered strain has huge economic and social benefits, and has a broad market development prospect.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a genetic engineering strain and a construction method thereof, in particular to a genetic engineering strain highly expressing α-glucosidase and a construction method thereof. Background technique [0002] α-glucosidase (α-transglucosidase, EC2.4.1.24), also known as α-glucosidase, transfer glucosidase, it can cut α-1 from the non-reducing end of the oligosaccharide substrate , 4 glycosidic bonds, release glucose, or transfer free glucose residues to another carbohydrate substrate to form α-1, 6 glycosidic bonds, thereby obtaining non-fermentable functional isomaltooligosaccharides (IMO), sugar lipids or glycopeptides. α-glucosidase is an important industrial enzyme, which has shown broad application prospects in food, feed, chemical industry, medicine and other fields, especially the huge application potential in IMO production has already aroused the attention of th...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/15C12N15/54C12N15/63C12R1/685
Inventor 黎明路福平刘萌
Owner TIANJIN UNIV OF SCI & TECH
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