Details zur Publikation
Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1128/AEM.01065-10
Titel (primär) Production of the chiral compound (R)-3-Hydroxybutyrate by a genetically engineered methylotrophic bacterium
Autor Hölscher, T.; Breuer, U.; Adrian, L.; Harms, H.; Maskow, T. ORCID logo
Quelle Applied and Environmental Microbiology
Erscheinungsjahr 2010
Department ISOBIO; UMB
Band/Volume 76
Heft 16
Seite von 5585
Seite bis 5591
Sprache englisch
Abstract In this study, a methylotrophic bacterium, Methylobacteriumrhodesianum MB 126, was used for the production of the chiral compound (R)-3-hydroxybutyrate (R-3HB) from methanol. R-3HB is formed during intracellular degradation of the storage polymer (R)-3-polyhydroxybutyrate (PHB). Since the monomer R-3HB does not accumulate under natural conditions, M. rhodesianum was genetically modified. The gene (hbd) encoding the R-3HB-degrading enzyme, R-3HB dehydrogenase, was inactivated in M. rhodesianum. The resulting hbd mutant still exhibited low growth rates on R-3HB as the sole source of carbon and energy, indicating the presence of alternative pathways for R-3HB utilization. Therefore, transposon mutagenesis was carried out with the hbd mutant, and a double mutant unable to grow on R-3HB was obtained. This mutant was shown to be defective in lipoic acid synthase (LipA), resulting in an incomplete citric acid cycle. Using the hbdlipA mutant, we produced 3.2 to 3.5 mM R-3HB in batch and 27 mM (2,800 mg liter-1) in fed-batch cultures. This was achieved by sequences of cultivation conditions initially favoring growth, then PHB accumulation, and finally PHB degradation.
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Hölscher, T., Breuer, U., Adrian, L., Harms, H., Maskow, T. (2010):
Production of the chiral compound (R)-3-Hydroxybutyrate by a genetically engineered methylotrophic bacterium
Appl. Environ. Microb. 76 (16), 5585 - 5591 10.1128/AEM.01065-10