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AssigneeJiangnan University
InventorsXueqin LV, Long LIU, Jian CHEN, Guocheng Du, Jianghua Li, Yanfeng LIU, Xiang XIU
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The invention provides a method for increasing the yield of 7-dehydrocholesterol in yeast by using compartmentalization. The method includes steps of: by taking yeast as a starting strain, expressing heterologous sterol delta 24-reductase and cholestenol delta-isomerase, wherein the yield of 7-DHC in Saccharomyces cerevisiae S288C is detected to be 10.15 mg/L. According to the method of the invention, partial enzymes in a 7-DHC synthesis path are positioned in compartments in Saccharomyces cerevisiae by using a peroxisome and a mitochondrial positioning tag, and a relatively independent 7-DHC synthesis path is formed. Meanwhile, the storage space of precursor substances needed by 7-DHC synthesis is increased, the feedback effect is reduced, the conversion efficiency between enzymes is improved in the same compartment, the loss of the acting substrate is reduced, and finally the yield of the 7-DHC is improved by 4 times and reaches 53.31 mg/L.
A method for increasing the yield of 7-dehydrocholesterol in yeast by using compartmentalization, comprising steps of:
A yeast engineered bacterium, wherein the yeast engineered bacterium expresses heterologous sterol delta 24-reductase and cholestenol delta-isomerase in peroxisomes and mitochondria of a yeast host respectively, and uses a peroxisome positioning tag and a mitochondrial positioning tag to express the enzymes in the 7-dehydrocholesterol synthesis pathway in peroxisomes and mitochondria respectively.
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