Enzymes expressed in saccharomyces cerevisiae, genetically engineered bacterium highly producing alpha- and gamma-tocotrienols as well as construction method of genetically engineered bacterium

Abstract

The invention discloses key enzymes expressed in saccharomyces cerevisiae, genetically engineered bacterium highly producing alpha- and gamma-tocotrienols as well as a construction method of the genetically engineered bacterium. Five enzymes capable of being expressed in the saccharomyces cerevisiae including HPPD, HPT, MPBQMT, TC and gamma-TMT respectively, are obtained through gene cloning and codon optimization; the five enzymes are integrated in saccharomyces cerevisiae genomes step by step, and the genetically engineered saccharomycete strain highly producing alpha- and gamma-tocotrienolsis constructed by further excising chloroplast transit peptides and over-expression rate-limiting enzyme, named YS-356c and has the collection number of CCTCC NO:M2019572. The tocotrienol yield reaches 2.09 mg / g dry cell weight. The engineered bacterium can be cultured by fermentation, alpha- and gamma-tocotrienols can be obtained directly from the bacterium, and the engineered bacterium has goodmarket prospects and application values.

Description

technical field [0001] The invention relates to the fields of genetic engineering and microorganisms, in particular to an enzyme expressed in Saccharomyces cerevisiae and a genetically engineered bacterium with high production of α- and γ-tocotrienol and a construction method thereof. Background technique [0002] Vitamin E is an important fat-soluble compound, including tocopherol and tocotrienol, which is an essential vitamin for maintaining normal metabolism and function of the body. Natural vitamin E only exists in photosynthetic organisms. Humans and animals cannot synthesize it by themselves and can only ingest it from the outside world. Vitamin E products on the market are mainly α-tocopherol, but recent studies have shown that compared to tocopherol, tocotrienol has the effect of lowering cholesterol (tocotrienol is unique), and better anti-oxidation, anti-cancer , anti-inflammatory, cardioprotective and neuroprotective functions. However, the sources of tocotrieno...

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Problems solved by technology

[0003] In 2008, researchers from the University of Stuttgart in Germany realized the synthesis of δ-tocotrienol in Escherichia coli by introducing genes from Arabidopsis and cyanobacteria, but the yield was very low, only 15 μg / g dry weight of cells, and δ- Tocotrienols are unmethylated forms of tocotrienols with limited applications

The invention provides a method for heterologously synthesizing α- and γ-tocotrienols with safe and efficient Saccharomyces cerevisiae as a cell factory, provides a new idea and method for industrial production of tocotrienols, and solves the problem of low output and production problems. high cost problem

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