Fermentation process capable of improving production level of recombinant human collagen

Abstract

The invention discloses a fermentation process capable of improving the production level of recombinant human collagen. The fermentation process comprises the following steps: firstly inoculating a Pichia pastoris strain liquid into a sterilized fermentation culture medium, fermentatively culturing for 14 to 18 hours, refilling methanol to carry out induction expression, simultaneously adding sodium pyruvate to the fermentation culture medium, wherein the sodium pyruvate is added in an amount of 0.01 to 10 g / L. In the fermentation process, the sodium pyruvate is added in the methanol-induced expression stage, so that the biosynthesis rate of the recombinant human collagen is improved; a continuous feed-batch way is adopted, so that the biosynthesis rate of the recombinant human collagen is further improved, and the fermentation time is shortened; meanwhile, the expression quantity of the recombinant human collagen is increased, the fermentation level is increased by more than 20%, and the production cost is reduced; the fermentation process is especially suitable for industrialized mass production of recombinant human collagen, and can bring huge practical application value for industrial production.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation and relates to a fermentation process for improving the production level of recombinant human collagen. Background technique [0002] Collagen is an important protein in animals. It is widely distributed in tissues such as skin, cartilage, and blood vessels. It participates in cell migration, differentiation, and reproduction, and plays an important role in maintaining the normal physiological functions of cells, tissues, and organs. It is used in food , Feed, beauty, cosmetics, medicine and other fields are widely used. At present, collagen raw materials are mainly obtained by separation and purification after treating animal skin, bone and other tissues such as pigs and cattle with physical and chemical methods such as acid, alkali and heating. However, the collagen obtained by the above method has complex components and poor water solubility, and because it comes from animal ti...

AI Technical Summary

Problems solved by technology

Compared with the traditional process, the fermentation process of genetically engineered bacteria of collagen has the advantages of easy availability of raw materials, environmental protection, and stable product quality, but there are also problems such as longer fermentation cycle and lower production efficiency.

Chinese patent 201010602214.8 discloses a production method for expressing recombinant human-like collagen in Pichia pastoris. Pichia pastoris C13 is used as the strain, the expression level of recombinant collagen is 5g / L, the fermentation period is 99 hours, and the fermentation level 0.051g / L·h, although the fermentation cycle is short, the protein expression and fermentation level are too low

Chinese patent 201110327865.5 constructed a Pichia pastoris genetically engineered strain of recombinant human collagen. The genetically engineered strain was fermented to obtain recombinant human collagen. The fermentation period was 136 hours, and the protein expression was 16g / L. The level is 0.118g / L·h, although the protein expression has increased, but the fermentation period is too long and the fermentation level is low