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Enhancing tylosin production by combinatorial overexpression of efflux,SAM biosynthesis,and regulatory genes in hyperproducing Streptomyces xinghaiensis strain

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Synthetic and Systems Biotechnology 2023年第3期8卷 486-497页

作者： Penghui Dai Yuyao Qin Luyuan Li Haidi Li Lihuo Lv Danying Xu Yuqing Song Tingting Huang Shuangjun Lin Zixin Deng Meifeng Tao State Key Laboratory of Microbial Metabolism School of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghai200240China Zhejiang Apeloa Biotechnology Co. Ltd.Jinhua322109China Zhejiang Apeloa Jiayuan Pharmaceutical Co. Ltd.Jinhua322118China Haihe Laboratory of Synthetic Biology Tianjin300308China

Tylosin is a 16-membered macrolide antibiotic widely used in veterinary medicine to control infections caused by Gram-positive pathogens and *** improve the fermentation titer of tylosin in the hyperproducing Streptomyces xinghaiensis strain TL01,we sequenced its whole genome and identified the biosynthetic gene cluster *** of the tylosin efflux gene tlrC,the cluster-situated S-adenosyl methionine(SAM)synthetase gene metK_(cs),the SAM biosynthetic genes adoK_(cs)-metFcs,or the pathway-specific activator gene tylR enhanced tylosin production by 18%,12%,11%,and 11%in the respective engineered strains TLPH08-2,TLPH09,TLPH10,and ***-overexpression of metK_(cs)and adoK_(cs)-metFcs as two transcripts increased tylosin production by 22%in the resultant strain TLPH11 compared to that in ***,combinational overexpression of tlrC,metK_(cs),adoK_(cs)-metFcs,and tylR as four transcripts increased tylosin production by 23%(10.93g/L)in the resultant strain TLPH17 compared to that in ***,a negligible additive effect was displayed upon combinational overexpression in TLPH17 as suggested by the limited increment of fermentation titer compared to that in *** analyses indicated that the expression of tlrC and three SAM biosynthetic genes in TLPH17 was considerably lower than that of TLPH08-2 and *** on this observation,the five genes were rearranged into one or two operons to coordinate their overexpression,yielding two engineered strains TLPH23 and TLPH24,and leading to further enhancement of tylosin production over *** particular,the production of TLPH23 reached 11.35 g/*** findings indicated that the combinatorial strategy is a promising approach for enhancing tylosin production in high-yielding industrial strains.

关键词： Streptomyces xinghaiensis Tylosin combinational metabolic engineering

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