Construction method of recombinant bacillus subtilis of high-yield pullulanase

A technology of Bacillus subtilis and pullulanase, which is applied in the field of construction of recombinant Bacillus subtilis and can solve problems such as no resistance
CN107058367APending Publication Date: 2017-08-18BAIYIN SINO BIOTECH
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
BAIYIN SINO BIOTECH
Publication Date
2017-08-18

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Abstract

The invention discloses a construction method of recombinant bacillus subtilis of high-yield pullulanase. The method comprises the steps that acid and heat resistance pullulanase gene is optimized by a codon, then is spiced artificially with two different secretory peptides and promoters to form a artificial operon and integrated into Bacillus subtilis A164; integration sites are the positions at which alkaline protease aprE and amylase gene amyE locate respectively; in addition, by the plasmid expression of Cre recombinase , resistance gene integrated into A164 is knocked down; pullulanase fermentation enzyme activity of pullulanase recombinant production bacterial strain CNP2 exceeds 1000 U / mL.
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Description

technical field

[0001] The invention belongs to the technical field of constructing safe production strains of enzymes used in the sugar industry, and relates to a method for constructing a high-yield pullulanase recombinant Bacillus subtilis. Background technique

[0002] Pullulanase is an indispensable enzyme in the sugar production technology of the food industry. It is mainly a sugar chain debranching enzyme secreted and expressed by prokaryotic microorganisms. Type I pullulanase in this type of enzyme is often used in production and application, which can specifically cut the branched chain branch composed of α-1,6 glycosidic bonds in starch raw materials, so pullulanase and other starch hydrolysis The combined action of enzymes can completely enzymatically hydrolyze starch to obtain single-molecule glucose molecules, which has broad market prospects in food industries such as starch processing. In recent years, a lot of manpower and material resources have been invest...

Claims

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