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PMID: 8300540 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Molecular characterization of an aldehyde/alcohol dehydrogenase gene from Clostridium acetobutylicum ATCC 824.

Journal of bacteriology ·Vol. 176 ·No. 3 ·1994-02-00 ·Pages 871-85

Nair RV, Bennett GN, Papoutsakis ET

Abstract

A gene (aad) coding for an aldehyde/alcohol dehydrogenase (AAD) was identified immediately upstream of the previously cloned ctfA (J. W. Cary, D. J. Petersen, E. T. Papoutsakis, and G. N. Bennett, Appl. Environ. Microbiol. 56:1576-1583, 1990) of Clostridium acetobutylicum ATCC 824 and sequenced. The 2,619-bp aad codes for a 96,517-Da protein. Primer extension analysis identified two transcriptional start sites 83 and 243 bp upstream of the aad start codon. The N-terminal section of AAD shows homology to aldehyde dehydrogenases of bacterial, fungal, mammalian, and plant origin, while the C-terminal section shows homology to alcohol dehydrogenases of bacterial (which includes three clostridial alcohol dehydrogenases) and yeast origin. AAD exhibits considerable amino acid homology (56% identity) over its entire sequence to the trifunctional protein encoded by adhE from Escherichia coli. Expression of aad from a plasmid in C. acetobutylicum showed that AAD, which appears as a approximately 96-kDa band in denaturing protein gels, provides elevated activities of NADH-dependent butanol dehydrogenase, NAD-dependent acetaldehyde dehydrogenase and butyraldehyde dehydrogenase, and a small increase in NADH-dependent ethanol dehydrogenase. A 957-bp open reading frame that could potentially encode a 36,704-Da protein was identified upstream of aad.

Related Genes
aad adh1 adhE ald aspni bdhA bdhB fucO l-ald mdh s-ald vibch
MeSH Terms
Alcohol Dehydrogenase/genetics Alcohol Oxidoreductases/genetics Aldehyde Dehydrogenase/genetics Aldehyde Oxidoreductases/genetics Amino Acid Sequence Bacterial Proteins Base Sequence Clostridium/enzymology,genetics Fermentation Gene Expression Genes, Bacterial Molecular Sequence Data Molecular Weight Multienzyme Complexes/genetics Promoter Regions, Genetic RNA, Messenger/genetics Restriction Mapping Sequence Alignment Sequence Homology, Amino Acid Transcription, Genetic
Chemicals
Bacterial Proteins Multienzyme Complexes RNA, Messenger AAD protein, Clostridium acetobutylicum Alcohol Oxidoreductases Alcohol Dehydrogenase Aldehyde Oxidoreductases Aldehyde Dehydrogenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nair R V
Department of Chemical Engineering, Northwestern University, Evanston, Illinois 60208.
Bennett G N
Papoutsakis E T
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.176.3.871-885.1994
Published
1994-02-00
Pages
871-85
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205125
Subset
IM
Databases
GENBANK
L14817
External Links
PubMed source DOI: 10.1128/jb.176.3.871-885.1994 PMC Full Text View journal details
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