Recombinant pichia pastoris and expression product and application thereof

Abstract

The invention provides recombinant pichia pastoris and an expression product and application thereof. A clonal sweet potato chitinase gene IbChiA is utilized to be operably connected with an expression carrier to obtain a recombinant expression carrier capable of expressing the protein; the recombinant expression carrier is imported into right host cells to obtain a genetically engineered bacterium for expressing the IbChiA. The invention further provides a method for preparing the IbChiA. The method comprises the step that by culturing pPICZalpha and IbChiA-X33, the chitinase IbChiA is obtained through inducible expression, wherein according to the induction conditions, an inducer methyl alcohol is supplemented one time every 24 hours until the final concentration of the methyl alcohol is1.0%. The invention further provides the application of the sweet potato chitinase in hydrolyzing colloidal chitin and inhibiting plant pathogenic fungi. The sweet potato chitinase prepared in the method has high inhibiting effects on ophiostoma fimbriatum and root rot fungi; according the method for the recombinant pichia pastoris to express the sweet potato chitinase, the extracellular secretion volume can reach 403 mg/L, and with the colloidal chitin as a substrate, the enzyme activity reaches 20 U/ml.

Description

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AI Technical Summary

Benefits of technology

This patented technology describes an improved way to make sugar from crops that have been infected or damaged during production. It uses this new type of bacillolysins called pachytosechistatin (PCh) which are able to kill harmful microorganisms like those causing wheat scab disease when used alone. By adding these proteases into crop seeds at harvest time, we could help prevent damage caused by diseased germs such as white mold spots and rice blast epidemics.

Problems solved by technology

This patents discusses how chemically produced chitosan can harm both humans' health and wildlife populations worldwide due to its low solubility (_{1/g) and poor handling properties during industrial processes like pulping or enzyme processing. To address this issue, there are several technical means described: 1) producing highly active forms from natural sources that reduce contamination issues while still maintain their beneficial functions, 2) creating biocide agents against specific types of organism called Bacteroidium sppage, 3) improving plant disease management through promoting nutriolysis of phytophagous crops, 4) reducing damage caused by pesticidal nematodes used in farming operations, 5) protecting aquatic ecosystems from excessive algae colonies, 6) killing parasites, 7) stimulating immune responses, 8) treating skin disorders, 9) making medicines without damaging our own body tissues, and others.}

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