title: L-serine production by a methylotroph and its related enzymes.
abstract: The production process of L-serine from methanol and glycine has been developed using a methylotroph with the serine pathway. Consecutive reactions of two enzymes, methanol dehydrogenase (MDH) and serine hydroxymethyltransferase (SHMT) are involved in the production. We screened a high producer, Hyphomicrobium methylovorum, which is an obligate methylotroph. With resting cells of the bacterium, 24 mg/ml of L-serine was produced from 100 mg/ml of glycine and 48 mg/ml of methanol in 3 days under optimal conditions. Next, a glycine-resistant mutant GM2 showed improved serine production (32-34 mg/ml). The mutant GM2 was found to have elevated activities of MDH and SHMT. Since there has so far been little report on the systematic characterization of enzymes of the serine pathway in methylotrophs, not only the above two enzymes but also the other three enzymes in H. methylovorum were purified and characterized: MDH, SHMT and hydroxypyruvate reductase (HPR) were crystallized; serine-glyoxylate aminotransferase (SGAT) and glycerate kinase (GK) were purified to homogeneity. As a result, all these enzymes were found to be stable against preservation and to exist abundantly in the bacterium. The gene of SHMT was cloned and its deduced amino acid sequence had homology to those of Escherichia coli (55%) and rabbit liver (44%), whereas the enzyme of the bacterium was immunochemically distinguishable from those of microorganisms other than Hyphomicrobium strains and mammalian livers.
authors.affiliation: Department of Biotechnology, Tottori University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Izumi
authors.forename: T
authors.initials: T
authors.lastname: Yoshida
authors.forename: S S
authors.initials: SS
authors.lastname: Miyazaki
authors.forename: T
authors.initials: T
authors.lastname: Mitsunaga
authors.forename: T
authors.initials: T
authors.lastname: Ohshiro
authors.forename: M
authors.initials: M
authors.lastname: Shimao
authors.forename: A
authors.initials: A
authors.lastname: Miyata
authors.forename: T
authors.initials: T
authors.lastname: Tanabe
mesh_terms: D000429:Alcohol Oxidoreductases / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics / Q000378:metabolism*; D001709:Biotechnology; D012696:Glycine Hydroxymethyltransferase / Q000235:genetics / Q000302:isolation & purification; D006038:Glyoxylates / Q000302:isolation & purification; D050537:Hydroxypyruvate Reductase; D008969:Molecular Sequence Data; D012694:Serine / Q000096:biosynthesis* / Q000302:isolation & purification; D000637:Transaminases / Q000302:isolation & purification
chemical_list: D006038:Glyoxylates; D012694:Serine; D000429:Alcohol Oxidoreductases; D050537:Hydroxypyruvate Reductase; C019641:alcohol dehydrogenase (acceptor); D012696:Glycine Hydroxymethyltransferase; D000637:Transaminases; C022636:serine-glyoxylate aminotransferase