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Applied Microbiology and Biotechnology

, Volume 39, Issue 4–5, pp 427–432 | Cite as

l-Serine production by a methylotroph and its related enzymes

  • Yoshikazu Izumi
  • Toyokazu Yoshida
  • Silvia Susana Miyazaki
  • Toshio Mitsunaga
  • Takashi Ohshiro
  • Masayuki Shimao
  • Atsuro Miyata
  • Tadashi Tanabe
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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.

Keywords

Enzyme Deduce Amino Acid Sequence Rabbit Liver Mammalian Liver Hyphomicrobium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Yoshikazu Izumi
    • 1
  • Toyokazu Yoshida
    • 2
  • Silvia Susana Miyazaki
    • 3
  • Toshio Mitsunaga
    • 2
  • Takashi Ohshiro
    • 1
  • Masayuki Shimao
    • 1
  • Atsuro Miyata
    • 4
  • Tadashi Tanabe
    • 4
  1. 1.Department of BiotechnologyTottori UniversityTottoriJapan
  2. 2.Department of Food and NutritionKinki UniversityNaraJapan
  3. 3.Department of Microbiology, Faculty of AgronomyBuenos Aires UniversityBuenos AiresArgentina
  4. 4.Department of PharmacologyNational Cardiovascular Center Research InstituteOsakaJapan

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