biostudies-literatureYang LHunan Provincial Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaChinese Ministry of Education 111 Project1982https://www.ebi.ac.uk/biostudies/studies/S-EPMC11085572biostudies-literatureUnknown29(9)Chalkophomycin is a novel chalkophore with antibiotic activities isolated from <i>Streptomyces</i> sp. CB00271, while its potential in studying cellular copper homeostasis makes it an important probe and drug lead. The constellation of <i>N</i>-hydroxylpyrrole, 2<i>H</i>-oxazoline, diazeniumdiolate, and methoxypyrrolinone functional groups into one compact molecular architecture capable of coordinating cupric ions draws interest to unprecedented enzymology responsible for chalkophomycin biosynthesis. To elucidate the biosynthetic machinery for chalkophomycin production, the <i>chm</i> biosynthetic gene cluster from <i>S</i>. sp. CB00271 was identified, and its involvement in chalkophomycin biosynthesis was confirmed by gene replacement. The <i>chm</i> cluster was localized to a ~31 kb DNA region, consisting of 19 open reading frames that encode five nonribosomal peptide synthetases (ChmHIJLO), one modular polyketide synthase (ChmP), six tailoring enzymes (ChmFGMNQR), two regulatory proteins (ChmAB), and four resistance proteins (ChmA'CDE). A model for chalkophomycin biosynthesis is proposed based on functional assignments from sequence analysis and structure modelling, and is further supported by analogy to over 100 <i>chm</i>-type gene clusters in public databases. Our studies thus set the stage to fully investigate chalkophomycin biosynthesis and to engineer chalkophomycin analogues through a synthetic biology approach.Molecules (Basel, Switzerland)Chalkophomycin Biosynthesis Revealing Unique Enzyme Architecture for a Hybrid Nonribosomal Peptide Synthetase and Polyketide Synthase.PMC1108557282373772BP08200342022JJ4040882204256Yang LGong BHuang YYi LChen LZhu XLi MDuan YfalseChalkophomycin Biosynthesis Revealing Unique Enzyme Architecture for a Hybrid Nonribosomal Peptide Synthetase and Polyketide Synthase.Chalkophomycin is a novel chalkophore with antibiotic activities isolated from <i>Streptomyces</i> sp. CB00271, while its potential in studying cellular copper homeostasis makes it an important probe and drug lead. The constellation of <i>N</i>-hydroxylpyrrole, 2<i>H</i>-oxazoline, diazeniumdiolate, and methoxypyrrolinone functional groups into one compact molecular architecture capable of coordinating cupric ions draws interest to unprecedented enzymology responsible for chalkophomycin biosynthesis. To elucidate the biosynthetic machinery for chalkophomycin production, the <i>chm</i> biosynthetic gene cluster from <i>S</i>. sp. CB00271 was identified, and its involvement in chalkophomycin biosynthesis was confirmed by gene replacement. The <i>chm</i> cluster was localized to a ~31 kb DNA region, consisting of 19 open reading frames that encode five nonribosomal peptide synthetases (ChmHIJLO), one modular polyketide synthase (ChmP), six tailoring enzymes (ChmFGMNQR), two regulatory proteins (ChmAB), and four resistance proteins (ChmA'CDE). A model for chalkophomycin biosynthesis is proposed based on functional assignments from sequence analysis and structure modelling, and is further supported by analogy to over 100 <i>chm</i>-type gene clusters in public databases. Our studies thus set the stage to fully investigate chalkophomycin biosynthesis and to engineer chalkophomycin analogues through a synthetic biology approach.2024-01-01T00:00:00Z2024 Apr2026-05-20T03:17:49.07Z2026-05-20T03:07:53.727ZS-EPMC110855723873147310.3390/molecules29091982