期刊
APPLIED BIOCHEMISTRY AND MICROBIOLOGY
卷 51, 期 2, 页码 249-254出版社
PLEIADES PUBLISHING INC
DOI: 10.1134/S0003683815020076
关键词
yeast Yarrowia lipolytica; threo-Ds-isocitric acid; rapeseed (canola) oil; mutagenesis; isocitrate liase; itaconic acid
资金
- Russian Foundation of Basic Research (RFBR)
- Government of Moscow oblast [14-48-03540]
- Ministry of Education and Science of the Russian Federation [RFMEFI62514X0005]
We studied the biosynthesis of isocitric acid from rapeseed (canola) oil by the yeast Yarrowia lipolytica and its regulation. We determined a fundamental possibility for directed biosynthesis of isocitric acid by Y. lipolytica yeast, with only minimal amounts of citric acid byproduct, when grown on a medium containing canola oil. Wild type strains of Y. lipolytica were mutagenized by UV irradiation and treatment with N-methyl-N'-nitro-N-nitrosoguanidine (NG). Subsequent selection on media with acetate and isocitrate resulted in isolation of a mutant Y. lipolytica UV/NG that synthesized isocitrate and citrate at a ratio of 2.7 : 1. In the parent strain, this ratio is 1 : 1. Inhibition of isocitrate lyase, a key enzyme in the metabolism of isocitric acid, by the addition of itaconic acid resulted in increased synthesis of isocitrate, with a ratio of isocitrate to citrate reaching 6 : 1. The cultivation of the Y. lipolytica UV/NG mutant in a pilot industrial fermenter in the presence of itaconic acid resulted in the production of 88.7 g/L of isocitric acid with a yield of 90%.
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