Preparation method for improving enzyme activity of pichia pastoris fermentation lipase and yield of methanol protein

Abstract

A preparation method for improving the enzyme activity of a pichia pastoris fermentation lipase and the yield of a methanol protein comprises the steps of: culture of primary seeds, culture of secondary seeds, high-density fermentation, induction for enzyme production, enzyme extraction, spray drying of enzymatic liquid and obtaining of the single-cell methanol protein; in the stage of induction for enzyme production, the seeds in the high-density fermentation stage are cultured until the dissolved oxygen rebounds, the seeds are continuously and hungrily cultured for 2-3 h; the seeds enter themethanol-polyol induction stage, the polyol is one of pentaerythritol, glycerin, trimethylolethane, xylitol and sorbitol; the polyol MPC is added to methanol in a volume ratio of 0.5-2:1, and the substances are fed together; the stirring speed is constant, methanol-polyol containing 12 mL / L of PTM1 is fed at a constant speed and is used for feeding induction; the concentration of methanol-polyolis increased in a gradient manner, from 1 mL / L / h-6 mL / L / h, and the concentration is increased by 1 mL / L / h per hour; the dissolved oxygen is controlled to be 10%-20%. With the preparation method provided by the invention, the enzyme activity of the pichia pastoris fermentation lipase can be improved and meanwhile the yield of the methanol protein is increased.

Description

technical field [0001] The invention relates to new material application technology, in particular to a preparation method for improving the enzyme activity of Pichia pastoris fermentation lipase and the yield of methanol protein. Background technique [0002] Lipase is a class of ester bond hydrolase that mainly hydrolyzes triglycerides formed from glycerol and water-insoluble long-chain fatty acids. It is widely used in food processing, new biomaterials, biomedicine, chiral drug resolution and other fields. The application of lipase in baked food can strengthen the gluten, improve the texture of dough and the quality of bread. The hydrolysis of milk fat by lipase can improve the quality of dairy products, and can be used to enhance the flavor of cheese and milk powder, cheese ripening, esterification modification of cream and ice cream, etc. Lipase is mainly used in the pharmaceutical industry to aid digestion and reduce blood fat, and can also be used for Clinical diagno...

AI Technical Summary

Problems solved by technology

However, the current production of methanol protein has the problems of single product, outdated fermentation process, high cost and low benefit; lipase, as a biological enzyme preparation, has mild reaction conditions and high catalytic efficiency, and is widely used in food, fermentation, tanning, Pharmaceutical, daily chemical, feed and other industries, however, at present, the lipase enzyme activity produced in the process of simultaneous joint fermentation of lipase and methanol protein is low and the output of methanol protein is low