Bacillomycin D is a cyclic lipopeptide produced by Bacillus amyloliquefaciens fmbJ.At present,no relevant report has described the combinatorial biosynthesis of bacillomycin D.Due to the strong biosynthetic potential ...Bacillomycin D is a cyclic lipopeptide produced by Bacillus amyloliquefaciens fmbJ.At present,no relevant report has described the combinatorial biosynthesis of bacillomycin D.Due to the strong biosynthetic potential of the communication-mediating(COM)domains,its crosstalk between NRPS subunits has been studied to some extent,but the interaction of COM domain between modules is rarely reported.Therefore,in this study,we conducted the combinatorial biosynthesis of bacillomycin D through the deletion of the COM donor and acceptor domains between the modules and elucidated the interaction between the NRPS modules.The results showed that the deletion of the donor domain between modules 2 and 3 did not affect catalysis by upstream modules,but prevented downstream modules from catalysing the extension of the lipopeptide product,ultimately resulting in mutant complexes that could form linear dipeptides with the sequenceβ-NH_(2)FA-Asn-Tyr.However,the engineered hybrid bacillomycin D NRPSs lacking the donor domains between modules 3 and 4 and modules 6 and 7 could form multiple assembly lines that produced bacillomycin D and its analogs(linear tripeptides,cyclic hexapeptides and linear hexapeptides).In addition,all the acceptor domain deletion strains failed to produce bacillomycin D,only truncated peptides produced by module interruption(except for the acceptor domain deletion strains between modules 3 and 4,which also produced cyclic hexapeptides).In conclusion,deletion of the inter-module donor domains led to a more flexible hybrid biosynthetic system for the production of diverse peptide products;compared with the inter-subunit donor domain deletion strains that could only produce truncated peptides,the former had a greater biosynthetic capacity.Meanwhile,the acceptor domains between modules were an important part of module-module interactions and efficient communication within bacillomycin D synthetase.展开更多
Bacillomycin D is a nonribosomal peptide produced by Bacillus amyloliquefaciens fmbJ.In this study,the biosynthesis of bacillomycin D was mediated by replacing the intermodule donor,acceptor,and communication-mediatin...Bacillomycin D is a nonribosomal peptide produced by Bacillus amyloliquefaciens fmbJ.In this study,the biosynthesis of bacillomycin D was mediated by replacing the intermodule donor,acceptor,and communication-mediating(COM)domain pairs and introducing the COM domain between the surfactin subunits.Both the homologous donor and the COM domain pair replacement strains eliminated the selective barrier of the original COM domain pair to a certain extent,resulting in a more flexible hybrid biosynthesis system that provides simultaneous biosynthesis of different lipopeptide products.The synthesis of bacillomycin D in the homologous acceptor replacement strains was barely affected.The COM domains between the surfactin subunits cannot establish efficient communication between the bacillomycin D modules.In conclusion,the COM domains compatibility between the bacillomycin D modules are extremely strong,and the conserved amino acid residues in the acceptor domain are an important part of module–module interactions and efficient communication during bacillomycin D synthesis.展开更多
Skeletonema costatum is a diatom widely distributed in red tide microalgae blooms and as one of the main algae causing harmful algal blooms,because of their rapid reproduction and production of toxic and harmful subst...Skeletonema costatum is a diatom widely distributed in red tide microalgae blooms and as one of the main algae causing harmful algal blooms,because of their rapid reproduction and production of toxic and harmful substances,often play a negative role in aquatic ecosystems,and human health and wellbeing.Bacillomycin D is a nonribosomal cyclic antifungal lipopeptide in the iturins family.In this study,Bacillomycin D was tested for its ability to inhibit the growth of S.costatum.The EC_(50 24h) of Bacillomycin D on S.costatum was 24.70μg/mL.The chlorophyll fluorescence parameters F_(v)/F_(m),F_(v)/F_(o),and yield of the diatoms decreased significantly with increasing concentrations of Bacillomycin D.Study of the mechanism showed that Bacillomycin D induced cell death by changing cell membrane permeability,promoting the release of cellular contents.In this study,transcriptomic analysis showed Bacillomycin D significantly inhibited the photosynthesis and metabolism of S.costatum.These findings investigated the inhibitory effect of Bacillomycin D on the growth of S.costatum and provided a theoretical foundation for the development of new environmentally friendly biological algicide.展开更多
基金supported by grants from the National Natural Science Foundation of China(grant no.31972174).
文摘Bacillomycin D is a cyclic lipopeptide produced by Bacillus amyloliquefaciens fmbJ.At present,no relevant report has described the combinatorial biosynthesis of bacillomycin D.Due to the strong biosynthetic potential of the communication-mediating(COM)domains,its crosstalk between NRPS subunits has been studied to some extent,but the interaction of COM domain between modules is rarely reported.Therefore,in this study,we conducted the combinatorial biosynthesis of bacillomycin D through the deletion of the COM donor and acceptor domains between the modules and elucidated the interaction between the NRPS modules.The results showed that the deletion of the donor domain between modules 2 and 3 did not affect catalysis by upstream modules,but prevented downstream modules from catalysing the extension of the lipopeptide product,ultimately resulting in mutant complexes that could form linear dipeptides with the sequenceβ-NH_(2)FA-Asn-Tyr.However,the engineered hybrid bacillomycin D NRPSs lacking the donor domains between modules 3 and 4 and modules 6 and 7 could form multiple assembly lines that produced bacillomycin D and its analogs(linear tripeptides,cyclic hexapeptides and linear hexapeptides).In addition,all the acceptor domain deletion strains failed to produce bacillomycin D,only truncated peptides produced by module interruption(except for the acceptor domain deletion strains between modules 3 and 4,which also produced cyclic hexapeptides).In conclusion,deletion of the inter-module donor domains led to a more flexible hybrid biosynthetic system for the production of diverse peptide products;compared with the inter-subunit donor domain deletion strains that could only produce truncated peptides,the former had a greater biosynthetic capacity.Meanwhile,the acceptor domains between modules were an important part of module-module interactions and efficient communication within bacillomycin D synthetase.
基金supported by grants from the National Natural Science Foundation of China(Grant No.31972174).
文摘Bacillomycin D is a nonribosomal peptide produced by Bacillus amyloliquefaciens fmbJ.In this study,the biosynthesis of bacillomycin D was mediated by replacing the intermodule donor,acceptor,and communication-mediating(COM)domain pairs and introducing the COM domain between the surfactin subunits.Both the homologous donor and the COM domain pair replacement strains eliminated the selective barrier of the original COM domain pair to a certain extent,resulting in a more flexible hybrid biosynthesis system that provides simultaneous biosynthesis of different lipopeptide products.The synthesis of bacillomycin D in the homologous acceptor replacement strains was barely affected.The COM domains between the surfactin subunits cannot establish efficient communication between the bacillomycin D modules.In conclusion,the COM domains compatibility between the bacillomycin D modules are extremely strong,and the conserved amino acid residues in the acceptor domain are an important part of module–module interactions and efficient communication during bacillomycin D synthesis.
基金financially supported by grants from the National Natural Science Foundation of China (No. 32100037,51971100)China Postdoctoral Science Foundation (No.2019M661767)+3 种基金Jiangsu Planned Projects for Postdoctoral Research Funds (No.2019K015)Natural Science Research General Project of Jiangsu Higher Education Institutions (No. 20KJB550008)Jiangsu Ocean University Research Funds (No. KQ17028)Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘Skeletonema costatum is a diatom widely distributed in red tide microalgae blooms and as one of the main algae causing harmful algal blooms,because of their rapid reproduction and production of toxic and harmful substances,often play a negative role in aquatic ecosystems,and human health and wellbeing.Bacillomycin D is a nonribosomal cyclic antifungal lipopeptide in the iturins family.In this study,Bacillomycin D was tested for its ability to inhibit the growth of S.costatum.The EC_(50 24h) of Bacillomycin D on S.costatum was 24.70μg/mL.The chlorophyll fluorescence parameters F_(v)/F_(m),F_(v)/F_(o),and yield of the diatoms decreased significantly with increasing concentrations of Bacillomycin D.Study of the mechanism showed that Bacillomycin D induced cell death by changing cell membrane permeability,promoting the release of cellular contents.In this study,transcriptomic analysis showed Bacillomycin D significantly inhibited the photosynthesis and metabolism of S.costatum.These findings investigated the inhibitory effect of Bacillomycin D on the growth of S.costatum and provided a theoretical foundation for the development of new environmentally friendly biological algicide.
