• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Nov 1992; 174(22): 7149–7158.
PMCID: PMC207405

Molecular characterization of two Clostridium acetobutylicum ATCC 824 butanol dehydrogenase isozyme genes.

Abstract

A 4-kb segment of DNA containing two previously cloned butanol dehydrogenase (BDH) isozyme genes (D. Petersen, R. Welch, F. Rudolph, and G. Bennett, J. Bacteriol. 173:1831-1834, 1991) was sequenced. Two complete open reading frames (ORFs) were identified (bdhA and bdhB), along with a third truncated ORF (ORF1). The translation products of bdhA and bdhB corresponded to the N-terminal sequences of the purified BDH I and BDH II proteins, respectively. The two isozymes had a high amino acid identity (73%) and showed homology to a newly described class of alcohol dehydrogenases. Northern blots revealed that bdhA and bdhB did not form an operon. Primer extension experiments located single transcriptional start sites 37 and 58 bp upstream of the start codons of bdhA and bdhB, respectively. The -10 and -35 promoter regions for these genes were almost identical. bdhA and bdhB were found to be induced or derepressed immediately prior to significant butanol production in controlled pH 5.0 batch fermentations.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bilofsky HS, Burks C. The GenBank genetic sequence data bank. Nucleic Acids Res. 1988 Mar 11;16(5):1861–1863. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Cary JW, Petersen DJ, Papoutsakis ET, Bennett GN. Cloning and expression of Clostridium acetobutylicum phosphotransbutyrylase and butyrate kinase genes in Escherichia coli. J Bacteriol. 1988 Oct;170(10):4613–4618. [PMC free article] [PubMed]
  • Collado-Vides J, Magasanik B, Gralla JD. Control site location and transcriptional regulation in Escherichia coli. Microbiol Rev. 1991 Sep;55(3):371–394. [PMC free article] [PubMed]
  • Conway T, Ingram LO. Similarity of Escherichia coli propanediol oxidoreductase (fucO product) and an unusual alcohol dehydrogenase from Zymomonas mobilis and Saccharomyces cerevisiae. J Bacteriol. 1989 Jul;171(7):3754–3759. [PMC free article] [PubMed]
  • Conway T, Sewell GW, Osman YA, Ingram LO. Cloning and sequencing of the alcohol dehydrogenase II gene from Zymomonas mobilis. J Bacteriol. 1987 Jun;169(6):2591–2597. [PMC free article] [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • DiLella AG, Woo SL. Cloning large segments of genomic DNA using cosmid vectors. Methods Enzymol. 1987;152:199–212. [PubMed]
  • Drewke C, Ciriacy M. Overexpression, purification and properties of alcohol dehydrogenase IV from Saccharomyces cerevisiae. Biochim Biophys Acta. 1988 May 6;950(1):54–60. [PubMed]
  • Gerischer U, Dürre P. mRNA analysis of the adc gene region of Clostridium acetobutylicum during the shift to solventogenesis. J Bacteriol. 1992 Jan;174(2):426–433. [PMC free article] [PubMed]
  • Goodlove PE, Cunningham PR, Parker J, Clark DP. Cloning and sequence analysis of the fermentative alcohol-dehydrogenase-encoding gene of Escherichia coli. Gene. 1989 Dec 21;85(1):209–214. [PubMed]
  • Janssen PJ, Jones DT, Woods DR. Studies on Clostridium acetobutylicum glnA promoters and antisense RNA. Mol Microbiol. 1990 Sep;4(9):1575–1583. [PubMed]
  • Jörnvall H, Persson B, Jeffery J. Characteristics of alcohol/polyol dehydrogenases. The zinc-containing long-chain alcohol dehydrogenases. Eur J Biochem. 1987 Sep 1;167(2):195–201. [PubMed]
  • Mackenzie KF, Eddy CK, Ingram LO. Modulation of alcohol dehydrogenase isoenzyme levels in Zymomonas mobilis by iron and zinc. J Bacteriol. 1989 Feb;171(2):1063–1067. [PMC free article] [PubMed]
  • Mermelstein LD, Welker NE, Bennett GN, Papoutsakis ET. Expression of cloned homologous fermentative genes in Clostridium acetobutylicum ATCC 824. Biotechnology (N Y) 1992 Feb;10(2):190–195. [PubMed]
  • Narberhaus F, Bahl H. Cloning, sequencing, and molecular analysis of the groESL operon of Clostridium acetobutylicum. J Bacteriol. 1992 May;174(10):3282–3289. [PMC free article] [PubMed]
  • Narberhaus F, Giebeler K, Bahl H. Molecular characterization of the dnaK gene region of Clostridium acetobutylicum, including grpE, dnaJ, and a new heat shock gene. J Bacteriol. 1992 May;174(10):3290–3299. [PMC free article] [PubMed]
  • Neale AD, Scopes RK, Kelly JM, Wettenhall RE. The two alcohol dehydrogenases of Zymomonas mobilis. Purification by differential dye ligand chromatography, molecular characterisation and physiological roles. Eur J Biochem. 1986 Jan 2;154(1):119–124. [PubMed]
  • Osman YA, Conway T, Bonetti SJ, Ingram LO. Glycolytic flux in Zymomonas mobilis: enzyme and metabolite levels during batch fermentation. J Bacteriol. 1987 Aug;169(8):3726–3736. [PMC free article] [PubMed]
  • Petersen DJ, Welch RW, Rudolph FB, Bennett GN. Molecular cloning of an alcohol (butanol) dehydrogenase gene cluster from Clostridium acetobutylicum ATCC 824. J Bacteriol. 1991 Mar;173(5):1831–1834. [PMC free article] [PubMed]
  • Petersen DJ, Bennett GN. Cloning of the Clostridium acetobutylicum ATCC 824 acetyl coenzyme A acetyltransferase (thiolase; EC 2.3.1.9) gene. Appl Environ Microbiol. 1991 Sep;57(9):2735–2741. [PMC free article] [PubMed]
  • Petersen DJ, Welch RW, Walter KA, Mermelstein LD, Papoutsakis ET, Rudolph FB, Bennett GN. Cloning of an NADH-dependent butanol dehydrogenase gene from Clostridium acetobutylicum. Ann N Y Acad Sci. 1991 Dec 27;646:94–98. [PubMed]
  • Rosenberg M, Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Villarroya A, Juan E, Egestad B, Jörnvall H. The primary structure of alcohol dehydrogenase from Drosophila lebanonensis. Extensive variation within insect 'short-chain' alcohol dehydrogenase lacking zinc. Eur J Biochem. 1989 Mar 1;180(1):191–197. [PubMed]
  • de Vries GE, Arfman N, Terpstra P, Dijkhuizen L. Cloning, expression, and sequence analysis of the Bacillus methanolicus C1 methanol dehydrogenase gene. J Bacteriol. 1992 Aug;174(16):5346–5353. [PMC free article] [PubMed]
  • Welch RW, Rudolph FB, Papoutsakis ET. Purification and characterization of the NADH-dependent butanol dehydrogenase from Clostridium acetobutylicum (ATCC 824). Arch Biochem Biophys. 1989 Sep;273(2):309–318. [PubMed]
  • Wiesenborn DP, Rudolph FB, Papoutsakis ET. Thiolase from Clostridium acetobutylicum ATCC 824 and Its Role in the Synthesis of Acids and Solvents. Appl Environ Microbiol. 1988 Nov;54(11):2717–2722. [PMC free article] [PubMed]
  • Williamson VM, Paquin CE. Homology of Saccharomyces cerevisiae ADH4 to an iron-activated alcohol dehydrogenase from Zymomonas mobilis. Mol Gen Genet. 1987 Sep;209(2):374–381. [PubMed]
  • Wills C, Kratofil P, Londo D, Martin T. Characterization of the two alcohol dehydrogenases of Zymomonas mobilis. Arch Biochem Biophys. 1981 Sep;210(2):775–785. [PubMed]
  • Wu L, Welker NE. Cloning and characterization of a glutamine transport operon of Bacillus stearothermophilus NUB36: effect of temperature on regulation of transcription. J Bacteriol. 1991 Aug;173(15):4877–4888. [PMC free article] [PubMed]
  • Yokoyama S, Yokoyama R, Kinlaw CS, Harry DE. Molecular evolution of the zinc-containing long-chain alcohol dehydrogenase genes. Mol Biol Evol. 1990 Mar;7(2):143–154. [PubMed]
  • Youngleson JS, Jones WA, Jones DT, Woods DR. Molecular analysis and nucleotide sequence of the adh1 gene encoding an NADPH-dependent butanol dehydrogenase in the Gram-positive anaerobe Clostridium acetobutylicum. Gene. 1989 May 30;78(2):355–364. [PubMed]
  • Youngleson Jonathan S, Santangelo Joseph D, Jones David T, Woods David R. Cloning and Expression of a Clostridium acetobutylicum Alcohol Dehydrogenase Gene in Escherichia coli. Appl Environ Microbiol. 1988 Mar;54(3):676–682. [PMC free article] [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...