Method for improving structural isomerism catalytic activity of CE enzyme and mutant thereof

A mutant and epimerase technology, applied in the field of enzyme engineering, can solve the problem of poor substrate affinity, which restricts the process of industrial application, and the catalytic properties of substrate affinity, lactulose conversion rate and thermal stability have not been greatly improved and other issues to achieve good catalytic performance
CN113564151AActive Publication Date: 2021-10-29JIANGNAN UNIV
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Patent Information

Authority / Receiving Office
CN ยท China
Patent Type
Applications(China)
Current Assignee / Owner
JIANGNAN UNIV
Publication Date
2021-10-29

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Abstract

The invention discloses a method for improving the structural isomerism catalytic activity of a CE enzyme and a mutant thereof, and belongs to the field of enzyme engineering. According to the invention, cellobiose epimerase (CsCE) derived from Caldicellosirutorschcharlyticus is taken as a research object, site-saturation mutagenesis is carried out on a site related to substrate binding based on a semi-rational strategy of sequence alignment and crystal structure analysis, a plurality of mutants with obviously improved structural isomerism catalytic activity are obtained, and the structural isomerism activity of the mutants is improved by about 36-232% than that of a wild type CE enzyme. The invention provides a feasible scheme for CE enzyme improvement, and has important significance for promoting enzymatic industrial synthesis of lactulose.
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Description

technical field

[0001] The invention relates to a method for improving the catalytic activity of CE enzyme structural isomerism and a mutant thereof, belonging to the field of enzyme engineering. Background technique

[0002] Lactulose (Lactulose, ฮฒ-D-galactosyl-1,4-D-fructose) is a non-digestible disaccharide, because of its ability to promote the growth of bifidobacteria, regulate intestinal pH, and inhibit the growth of pathogenic bacteria It is widely used in the food and pharmaceutical industries due to its prebiotic properties. At present, commercial lactulose on the market is mainly produced by chemical methods, which will produce high levels of by-products, and chemical methods involve complicated separation and purification steps, which will easily cause serious environmental pollution. These defects restrict the production of lactulose. application. Enzymatic synthesis of lactulose has become a research hotspot in the current production of lactulose because of it...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
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