Method for producing recombinant human-like collagen and host cell protein by using pichia pastoris

Abstract

The invention discloses a method for producing recombinant human-like collagen and host cell protein by using pichia pastoris, wherein the method comprises the steps: carrying out methanol induction on a strain Pichia pastoris-col3-6 in the fermentation culture process, and carrying out centrifugation, filtration sterilization, ultrafiltration desalination concentration, ion exchange chromatography, ultrafiltration desalination and freeze-drying on the obtained fermentation liquor to obtain the recombinant human-like collagen and the host cell protein. High yield, high yield and high purity of fermentation production of the recombinant human-like collagen are maintained, and meanwhile, the problem of high cost of industrial large-scale production of the recombinant human-like collagen expressed by pichia pastoris is solved.

Description

technical field [0001] The invention belongs to the technical field of protein engineering, and in particular relates to a method for separating and purifying recombinant human-like collagen expressed by Pichia pastoris and host cell protein. Background technique [0002] Collagen is the most abundant protein in the body, accounting for about 25% to 33% of human protein. It is widely distributed in various tissues and organs of the human body, such as skin, bones, cornea, blood vessels, etc., especially in skin and other tissues. Collagen, as a kind of adhesive substance of connective tissue, plays an important role in maintaining the normal physiological functions of cells, tissues and organs. Recombinant collagen is widely used in biomedicine, cosmetics, food and other industries due to its low toxicity, low antigenicity, low immunity, ability to guide cell regeneration and good biocompatibility. [0003] Protein Maker is made of a variety of purified proteins of known mo...

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Problems solved by technology

[0004] At present, during the purification process of recombinant human-like collagen expressed by Pichia pastoris, the production cost of recombinant human-like collagen is relatively high due to the investment in equipment, personnel and production operations.

At the same time, the host cell protein expressed by Pichia pastoris cannot be effectively utilized during the production process (the reason includes the limited content of the host cell protein in the fermentation broth, which is usually treated as an impurity in purification, for example, Chinese patent CN108027032A)

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