Activating and fermenting method of escherichia coli

Abstract

The invention is suitable for the technical field of microorganisms, and provides an activating and fermenting method of escherichia coli. The activating and fermenting method comprises the steps of taking escherichia coli, and performing inoculating in a culture medium to obtain a first-class flat; taking single colonies, continuing performing inoculating on the culture medium to obtain a second-class flat; taking many single colonies, performing inoculating on a liquid culture medium, and performing rotary culturing to obtain first-class seed liquid; continuing inoculating the liquid culturemedium with the first-class seed liquid, and performing rotary culturing to obtain second-class seed liquid; inoculating a fermentation base solution with the second-class seed liquid, performing fermentation, and supplementing foods in an oxygen dissolving regeneration manner; and adding an induction culture medium, inducing protein expression, taking constant-pH food supplementing for supplementing the foods, and continuing performing fermentation. According to the activating and fermenting method of escherichia coli provided by the embodiment of the invention, through flat culture twice and liquid culture twice, the second-class seed liquid is obtained, so that rejuvenation of strains can be effectively realized, the stability of a subsequent fermentation process can be effectively improved, and the expression level of target products can be increased effectively.

Description

technical field

[0001] The invention belongs to the technical field of microbes, and in particular relates to a method for activating and fermenting Escherichia coli. Background technique

[0002] Fermentation engineering is an important part of biotechnology and an important link in the industrialization of biotechnology. Fermentation technology has a long history. As a modern scientific concept, microbial fermentation industry is based on traditional fermentation technology, combined with modern new technologies such as genetic engineering and cell engineering. The fermentation process is often accompanied by protein expression. In the process of microbial fermentation, the activation of strains is a prerequisite to ensure the stability of fermentation and also affects the expression of target proteins, which is a crucial link.

[0003] However, the existing literature reports on the activation of strains often adopt the method of rejuvenating the strains, cultivating t...

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