Title |
Cloning, functional expression and characterization of a bifunctional 3-hydroxybutanal dehydrogenase /reductase involved in acetone metabolism by Desulfococcus biacutus
|
---|---|
Published in |
BMC Microbiology, November 2016
|
DOI | 10.1186/s12866-016-0899-9 |
Pubmed ID | |
Authors |
Jasmin Frey, Hendrik Rusche, Bernhard Schink, David Schleheck |
Abstract |
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 CO2 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. The MDR gene DebiaDRAFT_04514 was cloned and overexpressed in E. coli. The purified recombinant protein required zinc as cofactor, and accepted NADH/NAD(+) but not NADPH/NADP(+) 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 C3 - C5-aldehydes with NADH, such as propanal to propanol (380 ± 15 mU mg(-1) protein), butanal to butanol (300 ± 24 mU mg(-1)), and 3-hydroxybutanal to 1,3-butanediol (248 ± 60 mU mg(-1)), however, the enzyme also oxidized 3-hydroxybutanal with NAD(+) to acetoacetaldehyde (83 ± 18 mU mg(-1)). 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. |
Twitter Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Venezuela, Bolivarian Republic of | 1 | 50% |
Unknown | 1 | 50% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 50% |
Scientists | 1 | 50% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 9 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Lecturer | 2 | 22% |
Lecturer > Senior Lecturer | 1 | 11% |
Librarian | 1 | 11% |
Student > Bachelor | 1 | 11% |
Student > Ph. D. Student | 1 | 11% |
Other | 2 | 22% |
Unknown | 1 | 11% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 3 | 33% |
Environmental Science | 1 | 11% |
Immunology and Microbiology | 1 | 11% |
Earth and Planetary Sciences | 1 | 11% |
Social Sciences | 1 | 11% |
Other | 1 | 11% |
Unknown | 1 | 11% |