title: Carbon source dependent biosynthesis of acarviose metabolites in Actinoplanes sp. SE50/110.
abstract: In this work the biosynthesis of the type 2 diabetes mellitus therapeutic acarviosyl-maltose (acarbose) and related acarviose metabolites produced by Actinoplanes sp. SE50/110 was studied in liquid minimal medium supplemented with the defined carbon sources maltose, glucose, galactose or mixtures of maltose/glucose and maltose/galactose. Quantifying acarviosyl-maltose by HPLC and UV detection revealed that only cultures grown in maltose-containing minimal media produced acarviosyl-maltose in significant amounts. A qualitative analysis of the cytosolic and extracellular proteome for the presence of proteins from the acarbose biosynthesis gene cluster showed that these were not only synthesized in maltose-containing media, but also in media with glucose or galactose as the sole carbon source. A LC-MS-based detection method was applied to test the hypothesis that different acarviose metabolites are produced in media with maltose, glucose or galactose. The analysis revealed that a spectrum of acarviose metabolites (acarviose with 1-4 glucose equivalent units) was formed under all tested conditions. As expected, in maltose-containing minimal media acarviosyl-maltose was produced as the major component exceeding the remaining minor components by 2-3 orders of magnitude. In minimal medium supplemented with glucose acarviosyl-glucose was the major component, while in minimal medium with galactose no major component was present. Based on the results presented, a model for the intracellular biosynthesis of major and minor acarviose metabolites was developed.
authors.affiliation: Senior Research Group in Genome Research of Industrial Microorganisms, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany; CLIB-Graduate Cluster Industrial Biotechnology, CLIB(2021), Völklinger Strasse 4, 40219 Düsseldorf, Germany. Electronic address: Wendler@CeBiTec.Uni-Bielefeld.de.
authors.forename: Sergej
authors.initials: S
authors.lastname: Wendler
authors.affiliation: Senior Research Group in Genome Research of Industrial Microorganisms, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany; CLIB-Graduate Cluster Industrial Biotechnology, CLIB(2021), Völklinger Strasse 4, 40219 Düsseldorf, Germany; Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Vera
authors.initials: V
authors.lastname: Ortseifen
authors.affiliation: Senior Research Group in Genome Research of Industrial Microorganisms, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany; Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Persicke
authors.affiliation: Product Supply, Bayer Pharma AG, Friedrich Ebert Str. 217-475, 42117 Wuppertal, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Klein
authors.affiliation: CLIB-Graduate Cluster Industrial Biotechnology, CLIB(2021), Völklinger Strasse 4, 40219 Düsseldorf, Germany; Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Armin
authors.initials: A
authors.lastname: Neshat
authors.affiliation: Department of Proteome and Metabolome Research, Faculty of Biology, Bielefeld University, Universitätsstraße 25, 33615 Bielefeld, Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: Niehaus
authors.affiliation: Senior Research Group in Genome Research of Industrial Microorganisms, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Schneiker-Bekel
authors.affiliation: CLIB-Graduate Cluster Industrial Biotechnology, CLIB(2021), Völklinger Strasse 4, 40219 Düsseldorf, Germany; Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany; Department of Proteome and Metabolome Research, Faculty of Biology, Bielefeld University, Universitätsstraße 25, 33615 Bielefeld, Germany.
authors.forename: Frederik
authors.initials: F
authors.lastname: Walter
authors.affiliation: Department for Sports Medicine, University Wuppertal, Pauluskirchstr. 7, 42285 Wuppertal, Germany.
authors.forename: Udo F
authors.initials: UF
authors.lastname: Wehmeier
authors.affiliation: Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Jörn
authors.initials: J
authors.lastname: Kalinowski
authors.affiliation: Senior Research Group in Genome Research of Industrial Microorganisms, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Alfred
authors.initials: A
authors.lastname: Pühler
mesh_terms: D020909:Acarbose / Q000737:chemistry / Q000378:metabolism* / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D002244:Carbon / Q000737:chemistry*; D003924:Diabetes Mellitus, Type 2 / Q000188:drug therapy*; D005690:Galactose / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D016695:Glycosyltransferases / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008320:Maltose / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D008320:Maltose; D002244:Carbon; D016695:Glycosyltransferases; C432471:ACBD protein, Actinoplanes sp. 50-110; D005947:Glucose; D020909:Acarbose; D005690:Galactose
keywords: Acarbose; Acarviose metabolite; Actinoplanes; LC–MS; Proteome