Enhancement of erythritol production by Trichosporonoides oedocephalis ATCC 16958 through regulating key enzyme activity and the NADPH/NADP ratio with metal ion supplementation
2018
Li, Liangzhi | Kang, Pei | Ju, Xin | Chen, Jiajia | Zou, Huibin | Hu, Cuiying | Yan, Lishi
Erythritol, a well-known natural sweetener, is mainly produced by microbial fermentation. Various metal ions (Al³⁺, Cu²⁺, Mn²⁺, and Ni²⁺) were added to the culture medium of Trichosporonoides oedocephalis ATCC 16958 at 30 mg/L in shake flask cultures. Compared with controls, Cu²⁺ increased the erythritol content by 86% and decreased the glycerol by-product by 31%. After 48 hr of shake flask culture, sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that expression levels of erythrose reductase (ER) in the presence of 30 mg/L CuSO₄ · 5H₂O were higher than those obtained after treatment with other examined metal ions. Furthermore, after 108 hr of batch culture in a 5-L bioreactor, supplementation with 30 mg/L of CuSO₄ · 5H₂O increased the specific erythritol content by 27%. Further studies demonstrated that ER activity under 30 mg/L CuSO₄ · 5H₂O supplementation in a fermentor was overtly increased compared with the control after 60 hr, while glycerol-3-phosphate dehydrogenase activity was clearly reduced in most of the fermentation process. Furthermore, the NADPH/NADP ratio was slightly lower in T. oedocephalis cells treated with Cu²⁺ compared with control cells. These results provide further insights into Cu²⁺ effects on erythritol biosynthesis in T. oedocephalis and should improve the industrial production of erythritol by biological processes.
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