title: How To Make a Glycopeptide: A Synthetic Biology Approach To Expand Antibiotic Chemical Diversity.
abstract: Modification of natural product backbones is a proven strategy for the development of clinically useful antibiotics. Such modifications have traditionally been achieved through medicinal chemistry strategies or via in vitro enzymatic activities. In an orthogonal approach, engineering of biosynthetic pathways using synthetic biology techniques can generate chemical diversity. Here we report the use of a minimal teicoplanin class glycopeptide antibiotic (GPA) scaffold expressed in a production-optimized Streptomyces coelicolor strain to expand GPA chemical diversity. Thirteen scaffold-modifying enzymes from 7 GPA biosynthetic gene clusters in different combinations were introduced into S. coelicolor, enabling us to explore the criteria for in-cell GPA modification. These include identifying specific isozymes that tolerate the unnatural GPA scaffold and modifications that prevent or allow further elaboration by other enzymes. Overall, 15 molecules were detected, 9 of which have not been reported previously. Some of these compounds showed activity against GPA-resistant bacteria. This system allows us to observe the complex interplay between substrates and both non-native and native tailoring enzymes in a cell-based system and establishes rules for GPA synthetic biology and subsequent expansion of GPA chemical diversity.
authors.affiliation: M. G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University , Hamilton, Ontario L8S 4K1, Canada.
authors.forename: Grace
authors.initials: G
authors.lastname: Yim
authors.affiliation: M. G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University , Hamilton, Ontario L8S 4K1, Canada.
authors.forename: Wenliang
authors.initials: W
authors.lastname: Wang
authors.affiliation: M. G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University , Hamilton, Ontario L8S 4K1, Canada.
authors.forename: Maulik N
authors.initials: MN
authors.lastname: Thaker
authors.affiliation: M. G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University , Hamilton, Ontario L8S 4K1, Canada.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Tan
authors.affiliation: M. G. DeGroote Institute for Infectious Disease Research, Department of Biochemistry and Biomedical Sciences, McMaster University , Hamilton, Ontario L8S 4K1, Canada.
authors.forename: Gerard D
authors.initials: GD
authors.lastname: Wright
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D001419:Bacteria / Q000187:drug effects; D053898:Biosynthetic Pathways; D006020:Glycopeptides / Q000096:biosynthesis* / Q000737:chemistry; D005810:Multigene Family; D048372:Streptomyces coelicolor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058615:Synthetic Biology; D017334:Teicoplanin / Q000737:chemistry / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D006020:Glycopeptides; D017334:Teicoplanin
keywords: antibiotic; glycopeptide; heterologous expression; synthetic biosynthesis; tailoring enzymes