Pneumocandin B_(0)(PB_(0))is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate(Cancidas®).However,the low titer of PB_(0)result...Pneumocandin B_(0)(PB_(0))is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate(Cancidas®).However,the low titer of PB_(0)results in fermentation and purification costs during caspofungin production,limiting its widespread clinical application.Here,we engineered an efficient PB_(0)-producing strain of G.lozoyensis by systems metabolic engineering strategies,including multi-omics analysis and multilevel metabolic engineering.We overexpressed four rate-limiting enzymes:thioesterase GLHYD,two cytochrome P450s GLP450s,and chorismate synthase GLCS;knocked out two competing pathways responsible for producing 6-methylsalicylic acid and pyranidine E;and overexpressed the global transcriptional activator GLHYP.As a result,the PB_(0)titer increased by 108.7%to 2.63 g/L at the shake-flask level through combinatorial strategies.Our study provides valuable insights into achieving high-level production of PB_(0)and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.展开更多
基金funded by the National Key R&D Program of China(2021YFC2102600)the National Natural Science Foundation of China(32370060 and 32070063)the Shenzhen Science and Technology Program(JCYJ20241202124932044).
文摘Pneumocandin B_(0)(PB_(0))is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate(Cancidas®).However,the low titer of PB_(0)results in fermentation and purification costs during caspofungin production,limiting its widespread clinical application.Here,we engineered an efficient PB_(0)-producing strain of G.lozoyensis by systems metabolic engineering strategies,including multi-omics analysis and multilevel metabolic engineering.We overexpressed four rate-limiting enzymes:thioesterase GLHYD,two cytochrome P450s GLP450s,and chorismate synthase GLCS;knocked out two competing pathways responsible for producing 6-methylsalicylic acid and pyranidine E;and overexpressed the global transcriptional activator GLHYP.As a result,the PB_(0)titer increased by 108.7%to 2.63 g/L at the shake-flask level through combinatorial strategies.Our study provides valuable insights into achieving high-level production of PB_(0)and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.
