Molecular cloning of an alcohol (butanol) dehydrogenase gene cluster from Clostridium acetobutylicum ATCC 824.
In Clostridium acetobutylicum, conversion of butyraldehyde to butanol is enzymatically achieved by butanol dehydrogenase (BDH). A C. acetobutylicum gene that encodes this protein was identified by using an oligonucleotide designed on the basis of the N-terminal amino acid sequence of purified C. acetobutylicum NADH-dependent BDH. Enzyme assays of cell extracts of Escherichia coli harboring the clostridial gene demonstrated 15-fold-higher NADH-dependent BDH activity than untransformed E. coli, as well as an additional NADPH-dependent BDH activity. Kinetic, sequence, and isoelectric focusing analyses suggest that the cloned clostridial DNA contains two or more distinct C. acetobutylicum enzymes with BDH activity.
Petersen
D J
DJ
Department of Biochemistry and Cell Biology, William Marsh Rice University, Houston, Texas 77251.
Welch
R W
RW
Rudolph
F B
FB
Bennett
G N
GN
D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D007527:Isoenzymes / Q000235:genetics* / Q000302:isolation & purification; D008970:Molecular Weight; D005810:Multigene Family; D011994:Recombinant Proteins / Q000302:isolation & purification; D015183:Restriction Mapping
D007527:Isoenzymes; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; C060833:butanol dehydrogenase; D000426:Alcohol Dehydrogenase