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Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus

Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus

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Prüfsumme: MD5:83e412b0d22dad74927012452565091e

FREY, Jasmin, Hendrik RUSCHE, Bernhard SCHINK, David SCHLEHECK, 2016. Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus. In: BMC Microbiology. 16, 280. eISSN 1471-2180. Available under: doi: 10.1186/s12866-016-0899-9

@article{Frey2016-12Cloni-37677, title={Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus}, year={2016}, doi={10.1186/s12866-016-0899-9}, volume={16}, journal={BMC Microbiology}, author={Frey, Jasmin and Rusche, Hendrik and Schink, Bernhard and Schleheck, David}, note={Article Number: 280} }

Frey, Jasmin Schleheck, David Rusche, Hendrik Frey, Jasmin eng 2017-02-22T13:24:10Z Schink, Bernhard Schleheck, David Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus Background<br /><br />The strictly anaerobic, sulfate-reducing bacterium Desulfococcus biacutus can utilize acetone as sole carbon and energy source for growth. Whereas in aerobic and nitrate-reducing bacteria acetone is activated by carboxylation with CO<sub>2</sub> to acetoacetate, D. biacutus involves CO as a cosubstrate for acetone activation through a different, so far unknown pathway. Proteomic studies indicated that, among others, a predicted medium-chain dehydrogenase/reductase (MDR) superfamily, zinc-dependent alcohol dehydrogenase (locus tag DebiaDRAFT_04514) is specifically and highly produced during growth with acetone.<br /><br />Results<br /><br />The MDR gene DebiaDRAFT_04514 was cloned and overexpressed in E. coli. The purified recombinant protein required zinc as cofactor, and accepted NADH/NAD<sup>+</sup> but not NADPH/NADP<sup>+</sup> as electron donor/acceptor. The pH optimum was at pH 8, and the temperature optimum at 45 °C. Highest specific activities were observed for reduction of C<sub>3</sub> - C<sub>5</sub>-aldehydes with NADH, such as propanal to propanol (380 ± 15 mU mg<sup>−1</sup> protein), butanal to butanol (300 ± 24 mU mg<sup>−1</sup>), and 3-hydroxybutanal to 1,3-butanediol (248 ± 60 mU mg<sup>−1</sup>), however, the enzyme also oxidized 3-hydroxybutanal with NAD<sup>+</sup> to acetoacetaldehyde (83 ± 18 mU mg<sup>−1</sup>).<br /><br />Conclusion<br /><br />The enzyme might play a key role in acetone degradation by D. biacutus, for example as a bifunctional 3-hydroxybutanal dehydrogenase/reductase. Its recombinant production may represent an important step in the elucidation of the complete degradation pathway. Schink, Bernhard 2016-12 2017-02-22T13:24:10Z Rusche, Hendrik

Dateiabrufe seit 22.02.2017 (Informationen über die Zugriffsstatistik)

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