Abstract
For the conversion of glucose to 5-keto-d-gluconate (5-KGA), a precursor of the industrially important l-(+)-tartaric acid, Gluconobacter strains were genetically engineered. In order to increase 5-KGA formation, a plasmid-encoded copy of the gene encoding the gluconate:NADP-5 oxidoreductase (gno) was overexpressed in G. oxydans strain DSM 2434. This enzyme is involved in the nonphosphorylative ketogenic oxidation of glucose and oxidizes gluconate to 5-KGA. As the 5-KGA reductase activity depends on the cofactor NADP+, the sthA gene (encoding Escherichia coli transhydrogenase) was cloned and overexpressed in the GNO-overproducing G. oxydans strain. Growth of the sthA-carrying strains was indistinguishable from the G. oxydans wild-type strain and therefore they were chosen for the coupled overexpression of sthA and gno. G. oxydans strain DSM 2343/pRS201-gno-sthA overproducing both enzymes showed an enhanced accumulation of 5-KGA.
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This work was supported by the BMBF Verbund Mikrobielle Genomforschung (grant 031U213A) and the Fonds der Chemischen Industrie.
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Herrmann, U., Merfort, M., Jeude, M. et al. Biotransformation of glucose to 5-keto-d-gluconic acid by recombinant Gluconobacter oxydans DSM 2343. Appl Microbiol Biotechnol 64, 86–90 (2004). https://doi.org/10.1007/s00253-003-1455-8
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DOI: https://doi.org/10.1007/s00253-003-1455-8