title: Redox self-sufficient whole cell biotransformation for amination of alcohols.
abstract: Whole cell biotransformation is an upcoming tool to replace common chemical routes for functionalization and modification of desired molecules. In the approach presented here the production of various non-natural (di)amines was realized using the designed whole cell biocatalyst Escherichia coli W3110/pTrc99A-ald-adh-ta with plasmid-borne overexpression of genes for an l-alanine dehydrogenase, an alcohol dehydrogenase and a transaminase. Cascading alcohol oxidation with l-alanine dependent transamination and l-alanine dehydrogenase allowed for redox self-sufficient conversion of alcohols to the corresponding amines. The supplementation of the corresponding (di)alcohol precursors as well as amino group donor l-alanine and ammonium chloride were sufficient for amination and redox cofactor recycling in a resting buffer system. The addition of the transaminase cofactor pyridoxal-phosphate and the alcohol dehydrogenase cofactor NAD(+) was not necessary to obtain complete conversion. Secondary and cyclic alcohols, for example, 2-hexanol and cyclohexanol were not aminated. However, efficient redox self-sufficient amination of aliphatic and aromatic (di)alcohols in vivo was achieved with 1-hexanol, 1,10-decanediol and benzylalcohol being aminated best.
authors.affiliation: Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitaetsstr. 25, 33615 Bielefeld, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Klatte
authors.affiliation: Chair of Genetics of Prokaryotes, Faculty of Biology & CeBiTec, Bielefeld University, Universitaetsstr. 25, 33615 Bielefeld, Germany. Electronic address: volker.wendisch@uni-bielefeld.de.
authors.forename: Volker F
authors.initials: VF
authors.lastname: Wendisch
mesh_terms: D050788:Alanine Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D000438:Alcohols / Q000737:chemistry* / Q000378:metabolism*; D000586:Amination; D055162:Biocatalysis; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D010084:Oxidation-Reduction; D000637:Transaminases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000438:Alcohols; D000426:Alcohol Dehydrogenase; D050788:Alanine Dehydrogenase; D000637:Transaminases
keywords: Alanine dehydrogenase; Alcohol dehydrogenase; Cofactor recycling; Diamines; Escherichia coli; Redox self-sufficient amination; Transaminase; Whole cell biotransformation