Vitamin E is one of the most widely used vitamins.In the classical commercial synthesis of vitamin E(a-tocopherol),the chemical synthesis of isophytol is the key technical barrier.Here,we establish a new process for i...Vitamin E is one of the most widely used vitamins.In the classical commercial synthesis of vitamin E(a-tocopherol),the chemical synthesis of isophytol is the key technical barrier.Here,we establish a new process for isophytol synthesis from microbial fermented farnesene.To achieve an efficient pathway for farnesene production,Saccharomyces cerevisiae was selected as the host strain.First,b-farnesene synthase genes from different sources were screened,and through protein engineering and system metabolic engineering.展开更多
Metabolomics is an essential discipline in omics technology that promotes research on the biology of microbial systems.Streptomyces albus J1074 is a model organism used in fundamental research and industrial microbiol...Metabolomics is an essential discipline in omics technology that promotes research on the biology of microbial systems.Streptomyces albus J1074 is a model organism used in fundamental research and industrial microbiology.Nevertheless,a comprehensive and standardized method for analyzing the metabolome of S.albus J1074 is yet to be developed.Thus,we comprehensively evaluated and optimized the analytical procedure and sample preparation for profiling polar metabolites using hydrophilic interaction liquid chromatography(HILIC)coupled with high-resolution mass spectrometry(HRMS).We systematically examined the HILIC columns,quenching solutions,sample-to-quenching ratios,and extraction methods.Then,the optimal protocol was used to investigate the dynamic intracellular polar metabolite profile of the engineered S.albus J1074 strains during spinosad(spinosyn A and spinosyn D)fermentation.A total of 3648 compounds were detected,and 83 metabolites were matched to the standards.The intracellular metabolomic profiles of engineered S.albus J1074 strains(ADE-AP and OE3)were detected;furthermore,their metabolomes in different stages were analyzed to reveal the reasons for their differences in their spinosad production,as well as the current metabolic limitation of heterologous spinosad production in S.albus J1074.The HILIC-HRMS method is a valuable tool for investigating polar metabolomes,and provides a reference methodology to study other Streptomyces metabolomes.展开更多
基金This work was financially supported by the National Key R&D Program of China(2018YFA0900400)the National Natural Science Foundation of China(31670090).We also acknowledge Professor Zihe Liu fromthe Beijing University of Chemical Technology for kindly providing the pCas and KlURA3 plasmid.
文摘Vitamin E is one of the most widely used vitamins.In the classical commercial synthesis of vitamin E(a-tocopherol),the chemical synthesis of isophytol is the key technical barrier.Here,we establish a new process for isophytol synthesis from microbial fermented farnesene.To achieve an efficient pathway for farnesene production,Saccharomyces cerevisiae was selected as the host strain.First,b-farnesene synthase genes from different sources were screened,and through protein engineering and system metabolic engineering.
基金supported by the National Key R&D Program of China(grant number 2018YFA0900400)the National Natural Science Foundation of China(grant number 32100053).
文摘Metabolomics is an essential discipline in omics technology that promotes research on the biology of microbial systems.Streptomyces albus J1074 is a model organism used in fundamental research and industrial microbiology.Nevertheless,a comprehensive and standardized method for analyzing the metabolome of S.albus J1074 is yet to be developed.Thus,we comprehensively evaluated and optimized the analytical procedure and sample preparation for profiling polar metabolites using hydrophilic interaction liquid chromatography(HILIC)coupled with high-resolution mass spectrometry(HRMS).We systematically examined the HILIC columns,quenching solutions,sample-to-quenching ratios,and extraction methods.Then,the optimal protocol was used to investigate the dynamic intracellular polar metabolite profile of the engineered S.albus J1074 strains during spinosad(spinosyn A and spinosyn D)fermentation.A total of 3648 compounds were detected,and 83 metabolites were matched to the standards.The intracellular metabolomic profiles of engineered S.albus J1074 strains(ADE-AP and OE3)were detected;furthermore,their metabolomes in different stages were analyzed to reveal the reasons for their differences in their spinosad production,as well as the current metabolic limitation of heterologous spinosad production in S.albus J1074.The HILIC-HRMS method is a valuable tool for investigating polar metabolomes,and provides a reference methodology to study other Streptomyces metabolomes.