Cultivation method for saccharomycete excreting expression proteolytic enzyme

Abstract

The invention provides a method for culturing microzyme of secretory expression proteolytic enzyme. The method is characterized in that an inorganic salt culture medium is adopted for fermenting and culturing a recombinant methylotrophic yeast. Fermentation and culture comprise the growth period of the microzyme and the secretory expression period of the proteolytic enzyme. In the secretory expression period of the proteolytic enzyme, the feed flow of methanol can be controlled in real time according to the variation of oxygen dissolved in fermentation liquid, and the PH value of the fermentation liquid is adjusted with ammonia, thus expressing the proteolytic enzyme at high efficiency. Compared with the prior art, the method for culturing the microzyme of the secretory expression proteolytic enzyme has the characteristics of low culture cost, convenient and accurate reaction control mode and high expression activity of the proteolytic enzyme.

Description

Technical field: [0001] The invention relates to a yeast culture method for secreting and expressing protease. Background technique: [0002] Mannan-oligosaccharides and xylo-oligosaccharides are considered to be high-end products in functional oligosaccharides because of their good bifidus factor effects. Xylanase and mannanase are necessary biological enzymes for large-scale production of these two oligosaccharides. [0003] In order to obtain highly active xylanase and mannanase, people have tried various methods, especially applying genetic engineering means to obtain various recombinant strains to express secretase protein. In the published literature reports, Bacillus is more widely used. In recent years, some people have used Pichia pastoris (P. pastoris) to express mannanase and xylanase. For example, literature 1 uses the recombinant bacteria P. pastoris GS115-Atx to prepare xylanase. It is believed that the organic medium composed of bran and beef extract can be...

AI Technical Summary

Problems solved by technology

These methods, in the actual fermentation operation, are difficult to achieve real-time monitoring, there is a long delay between sampling and process adjustment, and the operation is cumbersome and the operating cost is relatively high, so it is difficult to achieve the best effect of the fermentation process

[0009] For the low-cost and efficient secretory expression of xylanase or mannanase, there is still a lack of corresponding technology