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Metabolic engineering of Gluconobacter oxydans 621H for increased biomass yield.
Appl Microbiol Biotechnol. 2017 Jul; 101(13):5453-5467.AM

Abstract

The obligatory aerobic acetic acid bacterium Gluconobacter oxydans incompletely oxidizes carbon sources regio- and stereoselectively in the periplasm and therefore is used industrially for oxidative biotransformations, e. g., in vitamin C production. However, it has a very low biomass yield as the oxidized products largely remain in the medium and cannot be used for anabolism. Cytoplasmic carbon metabolism occurs via the pentose phosphate pathway and the Entner-Doudoroff pathway, whereas glycolysis and the tricarboxylic acid cycle are incomplete. Acetate is formed as an end product via pyruvate decarboxylase and acetaldehyde dehydrogenase. In order to increase the biomass yield from glucose, we sequentially replaced (i) gdhS encoding the cytoplasmic NADP-dependent glucose dehydrogenase by the Acetobacter pasteurianus sdhCDABE genes for succinate dehydrogenase and the flavinylation factor SdhE (strain IK001), (ii) pdc encoding pyruvate decarboxylase by a second ndh gene encoding a type II NADH dehydrogenase (strain IK002.1), and (iii) gdhM encoding the membrane-bound PQQ-dependent glucose dehydrogenase by sucCD from Gluconacetobacter diazotrophicus encoding succinyl-CoA synthetase (strain IK003.1). Analysis of the strains under controlled cultivation conditions in bioreactors revealed for IK003.1 that neither gluconate nor 2-ketogluconate was formed, but some 5-ketogluconate. Acetate formation was eliminated, and comparable amounts of pyruvate were formed instead. CO2 formation by IK003.1 was more than doubled compared to the reference strain. Growth of IK003.1 was retarded, but the biomass yield of this strain was raised by 60%. IK003.1 serves as suitable host for oxidative biotransformations and for further metabolic engineering.

Authors+Show Affiliations

IBG-1: Biotechnology, Institute of Bio- and Geosciences, Forschungszentrum Jülich, Wilhelm-Johnen-Strasse, D-52425, Jülich, Germany. The Bioeconomy Science Center (BioSC), Forschungszentrum Jülich, D-52425, Jülich, Germany.IBG-1: Biotechnology, Institute of Bio- and Geosciences, Forschungszentrum Jülich, Wilhelm-Johnen-Strasse, D-52425, Jülich, Germany. The Bioeconomy Science Center (BioSC), Forschungszentrum Jülich, D-52425, Jülich, Germany.IBG-1: Biotechnology, Institute of Bio- and Geosciences, Forschungszentrum Jülich, Wilhelm-Johnen-Strasse, D-52425, Jülich, Germany. m.bott@fz-juelich.de. The Bioeconomy Science Center (BioSC), Forschungszentrum Jülich, D-52425, Jülich, Germany. m.bott@fz-juelich.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28484812

Citation

Kiefler, Ines, et al. "Metabolic Engineering of Gluconobacter Oxydans 621H for Increased Biomass Yield." Applied Microbiology and Biotechnology, vol. 101, no. 13, 2017, pp. 5453-5467.
Kiefler I, Bringer S, Bott M. Metabolic engineering of Gluconobacter oxydans 621H for increased biomass yield. Appl Microbiol Biotechnol. 2017;101(13):5453-5467.
Kiefler, I., Bringer, S., & Bott, M. (2017). Metabolic engineering of Gluconobacter oxydans 621H for increased biomass yield. Applied Microbiology and Biotechnology, 101(13), 5453-5467. https://doi.org/10.1007/s00253-017-8308-3
Kiefler I, Bringer S, Bott M. Metabolic Engineering of Gluconobacter Oxydans 621H for Increased Biomass Yield. Appl Microbiol Biotechnol. 2017;101(13):5453-5467. PubMed PMID: 28484812.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Metabolic engineering of Gluconobacter oxydans 621H for increased biomass yield. AU - Kiefler,Ines, AU - Bringer,Stephanie, AU - Bott,Michael, Y1 - 2017/05/08/ PY - 2017/03/16/received PY - 2017/04/24/accepted PY - 2017/04/20/revised PY - 2017/5/10/pubmed PY - 2018/1/5/medline PY - 2017/5/10/entrez KW - Biomass yield KW - Genetic engineering KW - Gluconobacter oxydans KW - Glucose dehydrogenase KW - Succinate dehydrogenase KW - Succinyl-CoA synthetase KW - Tricarboxylic acid cycle SP - 5453 EP - 5467 JF - Applied microbiology and biotechnology JO - Appl. Microbiol. Biotechnol. VL - 101 IS - 13 N2 - The obligatory aerobic acetic acid bacterium Gluconobacter oxydans incompletely oxidizes carbon sources regio- and stereoselectively in the periplasm and therefore is used industrially for oxidative biotransformations, e. g., in vitamin C production. However, it has a very low biomass yield as the oxidized products largely remain in the medium and cannot be used for anabolism. Cytoplasmic carbon metabolism occurs via the pentose phosphate pathway and the Entner-Doudoroff pathway, whereas glycolysis and the tricarboxylic acid cycle are incomplete. Acetate is formed as an end product via pyruvate decarboxylase and acetaldehyde dehydrogenase. In order to increase the biomass yield from glucose, we sequentially replaced (i) gdhS encoding the cytoplasmic NADP-dependent glucose dehydrogenase by the Acetobacter pasteurianus sdhCDABE genes for succinate dehydrogenase and the flavinylation factor SdhE (strain IK001), (ii) pdc encoding pyruvate decarboxylase by a second ndh gene encoding a type II NADH dehydrogenase (strain IK002.1), and (iii) gdhM encoding the membrane-bound PQQ-dependent glucose dehydrogenase by sucCD from Gluconacetobacter diazotrophicus encoding succinyl-CoA synthetase (strain IK003.1). Analysis of the strains under controlled cultivation conditions in bioreactors revealed for IK003.1 that neither gluconate nor 2-ketogluconate was formed, but some 5-ketogluconate. Acetate formation was eliminated, and comparable amounts of pyruvate were formed instead. CO2 formation by IK003.1 was more than doubled compared to the reference strain. Growth of IK003.1 was retarded, but the biomass yield of this strain was raised by 60%. IK003.1 serves as suitable host for oxidative biotransformations and for further metabolic engineering. SN - 1432-0614 UR - https://www.unboundmedicine.com/medline/citation/28484812/Metabolic_engineering_of_Gluconobacter_oxydans_621H_for_increased_biomass_yield_ L2 - https://dx.doi.org/10.1007/s00253-017-8308-3 DB - PRIME DP - Unbound Medicine ER -