title: In vitro characterization of LmbK and LmbO: identification of GDP-D-erythro-α-D-gluco-octose as a key intermediate in lincomycin A biosynthesis.
abstract: Lincomycin A is a clinically useful antibiotic isolated from Streptomyces lincolnensis. It contains an unusual methylmercapto-substituted octose, methylthiolincosamide (MTL). While it has been demonstrated that the C8 backbone of MTL moiety is derived from D-fructose 6-phosphate and D-ribose 5-phosphate via a transaldol reaction catalyzed by LmbR, the subsequent enzymatic transformations leading to the MTL moiety remain elusive. Here, we report the identification of GDP-D-erythro-α-D-gluco-octose (GDP-D-α-D-octose) as a key intermediate in the MTL biosynthetic pathway. Our data show that the octose 1,8-bisphosphate intermediate is first converted to octose 1-phosphate by a phosphatase, LmbK. The subsequent conversion of the octose 1-phosphate to GDP-D-α-D-octose is catalyzed by the octose 1-phosphate guanylyltransferase, LmbO. These results provide significant insight into the lincomycin biosynthetic pathway, because the activated octose likely serves as the acceptor for the installation of the C1 sulfur appendage of MTL.
authors.affiliation: Division of Medicinal Chemistry, College of Pharmacy, and Department of Chemistry, University of Texas at Austin , Austin, Texas 78712, United States.
authors.forename: Chia-I
authors.initials: CI
authors.lastname: Lin
authors.forename: Eita
authors.initials: E
authors.lastname: Sasaki
authors.forename: Aoshu
authors.initials: A
authors.lastname: Zhong
authors.forename: Hung-wen
authors.initials: HW
authors.lastname: Liu
mesh_terms: D006156:Guanosine Diphosphate Sugars / Q000378:metabolism*; D008034:Lincomycin / Q000096:biosynthesis*; D009005:Monosaccharides / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology
chemical_list: D006156:Guanosine Diphosphate Sugars; D009005:Monosaccharides; D008034:Lincomycin; D009713:Nucleotidyltransferases; C020842:guanylyltransferase; D010744:Phosphoric Monoester Hydrolases