Method for preparing high temperature-resistant lipase

Abstract

The invention discloses a method for preparing high temperature-resistant lipase. The method comprise the steps of cloning high temperature-resistant lipase genes coming from Bacillus subtilis into pichia pastoris; performing activating treatment on pichia pastoris, and then inoculating pichia pastoris having undergone the activating treatment into a shake flask to perform shake-flask culture; transferring bacterial solution undergoing the shake-flask culture to a seed tank to perform culture; transferring bacterial solution cultured in the seed tank to a fermentation tank to perform fermentation culture; finally adding lipase synergist in fermentation broth, and obtaining the lipase after adsorption and drying. The lipase obtained according to the preparation method has the advantages of resisting high temperature, having a wide pH action sphere and having good tolerance to organic solvents, the applied range of the lipase is expanded, and the problem that the lipase is not resistant to high temperature in feed granulation is solved.

Description

technical field [0001] The invention relates to the field of fermentation, in particular to a preparation method of high temperature resistant lipase. Background technique [0002] Lipase (triacylglycerollipase EC3.1.1.3) is a ubiquitous hydrolase that can catalyze the breaking of triglyceride bonds. Because of its diversity of catalytic functions and its superiority as a biocatalyst, it is widely used in food, tanning, feed, washing, oleochemical and other fields. As a feed additive, lipase has broad application prospects in animal breeding. First, lipase can eliminate lipid anti-nutritional factors in plant-based feed materials, and at the same time, it can make up for the deficiency of lipase in the digestive tract of poultry and young animals. However, in the lipase products that have been industrialized, the pH stability is mostly in the range of neutral and alkaline, and the tolerance to acid is poor, and there are few varieties suitable for feed. [0003] The prepar...

AI Technical Summary

Problems solved by technology

Plants such as the seeds of oil crops, animals such as the pancreas and adipose tissue of higher animals are the source of lipase extraction method, the extraction method has limited resources, complicated process, and low yield; microbial fermentation method is not affected by the environment, resources Abundant, short enzyme production cycle, relatively simple product and low cost, easy to manage in production, easy to realize industrial production, but the temperature resistance of lipase prepared by microbial fermentation method is relatively poor

[0004] Bacillus is an important member of lipase-producing microorganisms. The lipase produced by it has the characteristics of high temperature tolerance, wide pH range and good tolerance to organic solvents. However, under natural conditions , the enzyme yield of the bacteria is low, and it is difficult to meet the requirements of industrial production by conventional breeding methods

If it is possible to study a new preparation method using lipase, it is necessary to overcome the low temperature resistance of the lipase prepared by microbial fermentation in the prior art and to ensure the industrial production of the preparation method.

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