A method for non-methanol-induced production of antimicrobial peptides

Abstract

The invention discloses a method for producing antibacterial peptide through non-methanol induction. The invention relates to the technical field of the bioengineering and bio-pharmaceuticals, the bad expression effect of the antibacterial peptide in the pichia pastoris is solved, the most important is the methanol resides in the antibacterial peptide product expressed by use of the methanol induction, the toxicity of the methanol enables the antibacterial peptide product to be not applied in the fields of the food, the fodder and the medicines. The technical scheme provided by the invention is to add the inductor choline chloride into an inoculated culture medium so that the final concentration of the choline chloride in the culture medium is 1%-5%w / v. The method disclosed by the invention has the advantages that the PisL9K22WK expression level is high, the production cost is low, the product is safe and free from poisonous residue; the engineering saccharomycetes and the produced pisL9K22WK product can be applied to the antibacterial medicines or the animal or aquiculture fodder additives.

Description

technical field [0001] The invention relates to the technical fields of bioengineering and biopharmaceuticals, and more particularly relates to a method for inducing the high-efficiency expression of the improved grouper antimicrobial peptide pisL9K22WK in Pichia pink by using a non-toxic inducer. Background technique [0002] Antimicrobial peptides (AMP) are positively charged, cation-rich, and amphiphilic small molecule polypeptides in organisms. The main characteristics are as follows: high antibacterial activity, wide antibacterial spectrum, and many types. It is not easy to produce drug resistance to bacteria. The post-metabolism of the body is fast, it is not easy to produce harmful residues in the body, and it has killing or inhibiting effects on fungi, viruses, protozoa and even tumors. Due to its outstanding biological characteristics, it has the most potential as a new type of safe and harmless antibiotic substitute, and has attracted the attention of the scientifi...

AI Technical Summary

Problems solved by technology

[0006] However, the current Pichia pastoris expression system faces many problems in the fermentation and amplification process: methanol is used as a carbon source when inducing the expression of exogenous recombinant proteins. Methanol is toxic, not suitable for the production of food or additives, and is flammable and explosive. It is easy to be volatile, and the workshop needs to be anti-riot design in industrial production, which leads to increased costs; methanol fermentation consumes a lot of oxygen, and it is generally difficult to meet the demand for oxygen by increasing the air volume and speed of the air, so it is necessary to pass a lot of pure oxygen. Industrialized production brings inconvenience. In addition, the more methanol consumed, the greater the heat produced, and the higher the cooling capacity of the required equipment; the efficient transcription of the Pichia pastoris AOX1 promoter cannot do without methanol, which suffers from other carbon sources. carbon source repression

[0007] These problems lead to poor expression of antimicrobial peptides in Pichia pastoris. More importantly, methanol is retained in the antimicrobial peptide products induced by methanol. The toxicity of methanol makes this antimicrobial peptide product unable to be used in the fields of food, feed and medicine.

Images