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Enhanced lincomycin A production by calcium gluconate feeding in fermentation of Streptomyces lincolnensis

Enhanced lincomycin A production by calcium gluconate feedin...

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中国生物工程学会第十三届学术年会暨2019年全国生物技术大会

作者： Zhihui Zhuang Liwen Zhang Cancan Yang Deyu Zhu Quangui Mao Qiyao Wang Shuhong Gao State Key Laboratory of Bioreactor Engineering East China University of Science and Technology Topfond Pharmaceutical Co. Ltd

Lincomycin A is a clinically important antibiotic produced by Streptomyces lincolnensis that is used against gram-positive bacteria. To increase the yield of lincomycin A, a calcium gluconate feeding strategy was studied in a 15 L bioreactor. The results showed that the addition time of calcium gluconate was optimal during the late fermentation process to ensure a higher yield of lincomycin A. The optimum addition was continuous feeding at a speed of 0.0638 g/L/h from 111 to 158 h, which can increase the lincomycin A titer to 9160 mg/L, 41.3% higher than that without gluconate feeding. Enzyme activities of the central metabolic pathways, accumulation of intermediate metabolites, nadph and nadh concentrations, and nadph/nadh ratio were determined to investigate the mechanism of enhanced lincomycin A production by calcium gluconate addition. The activities of key enzymes of the pentose phosphate pathway(PPP)(glucose 6-phosphate dehydrogenase) and TCA cycle(isocitrate dehydrogenase) were enhanced by approximately twofold. A higher ratio of nadph/nadh was observed in the fermentation process with the optimized feeding strategy providing su cient reducing power. These data indicated that more flux flows through the PPP and the TCA cycle to provide more precursors, ATP and reducing power to support the synthesis of lincomycin A. The results showed a new strategy to improve the production of lincomycin A by manipulating the flux through the PPP and the TCA cycle.

关键词： Streptomyces lincolnensis Lincomycin Calcium gluconate nadph/nadh ratio

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