Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. 1995 Sep; 177(18): 5261–5269.
PMCID: PMC177317

Identification of the transcriptional activator controlling the butanediol fermentation pathway in Klebsiella terrigena.


The gene budR, whose product is responsible for induction of the butanediol formation pathway under fermentative growth conditions in Klebsiella terrigena, has been cloned and sequenced. This gene is separated from the budABC operon by a nontranslated region of 106 bp and transcribed in the opposite direction. budR codes for a protein of molecular weight 32,124, the sequence of which exhibits characteristics of regulators belonging to the LysR family. When transferred into the heterologous host Escherichia coli, budR activates expression of budA'-lacZ transcriptional and translational fusions with a regulatory pattern identical to that in K. terrigena, namely, induction by acetate, low pH, and anaerobiosis. Induction by acetate was specific, indicating that it is the physiological inducer. Primer extension analysis located the start site of transcription to two positions, 23 and 24 bp upstream of the budR initiation codon, and also showed that BudR strongly autoregulates its own expression. The products of fhlA, arcA, hip, ntrA, and katF did not influence expression of the bud operon. A mutation in fnr, however, led to a threefold increase in expression, indicating that Fnr acts as a repressor. The results support the notion that BudR coordinates the activity of the energy-conserving, nonreductive, but acidifying acetate formation pathway with the expression of the non-energy-conserving, reductive, but nonacidifying butanediol pathway.

Full Text

The Full Text of this article is available as a PDF (381K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aiba H, Adhya S, de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed]
  • Birkmann A, Sawers RG, Böck A. Involvement of the ntrA gene product in the anaerobic metabolism of Escherichia coli. Mol Gen Genet. 1987 Dec;210(3):535–542. [PubMed]
  • Birkmann A, Zinoni F, Sawers G, Böck A. Factors affecting transcriptional regulation of the formate-hydrogen-lyase pathway of Escherichia coli. Arch Microbiol. 1987 Jun;148(1):44–51. [PubMed]
  • Blomqvist K, Nikkola M, Lehtovaara P, Suihko ML, Airaksinen U, Stråby KB, Knowles JK, Penttilä ME. Characterization of the genes of the 2,3-butanediol operons from Klebsiella terrigena and Enterobacter aerogenes. J Bacteriol. 1993 Mar;175(5):1392–1404. [PMC free article] [PubMed]
  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed]
  • Brown TD, Pereira CR, Stormer FC. Studies of the acetate kinase-phosphotransacetylase and the butanediol-forming systems in Aerobacter aerogenes. J Bacteriol. 1972 Dec;112(3):1106–1111. [PMC free article] [PubMed]
  • Casadaban MJ, Cohen SN. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. [PMC free article] [PubMed]
  • Chen YM, Lin EC. Regulation of the adhE gene, which encodes ethanol dehydrogenase in Escherichia coli. J Bacteriol. 1991 Dec;173(24):8009–8013. [PMC free article] [PubMed]
  • Green J, Guest JR. Regulation of transcription at the ndh promoter of Escherichia coli by FNR and novel factors. Mol Microbiol. 1994 May;12(3):433–444. [PubMed]
  • Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. [PubMed]
  • Henikoff S, Haughn GW, Calvo JM, Wallace JC. A large family of bacterial activator proteins. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6602–6606. [PMC free article] [PubMed]
  • Holmes DS, Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. [PubMed]
  • Iuchi S, Furlong D, Lin EC. Differentiation of arcA, arcB, and cpxA mutant phenotypes of Escherichia coli by sex pilus formation and enzyme regulation. J Bacteriol. 1989 May;171(5):2889–2893. [PMC free article] [PubMed]
  • Iuchi S, Lin EC. Adaptation of Escherichia coli to redox environments by gene expression. Mol Microbiol. 1993 Jul;9(1):9–15. [PubMed]
  • Johansen L, Bryn K, Stormer FC. Physiological and biochemical role of the butanediol pathway in Aerobacter (Enterobacter) aerogenes. J Bacteriol. 1975 Sep;123(3):1124–1130. [PMC free article] [PubMed]
  • Kleckner N, Barker DF, Ross DG, Botstein D. Properties of the translocatable tetracycline-resistance element Tn10 in Escherichia coli and bacteriophage lambda. Genetics. 1978 Nov;90(3):427–461. [PMC free article] [PubMed]
  • Lange R, Hengge-Aronis R. Identification of a central regulator of stationary-phase gene expression in Escherichia coli. Mol Microbiol. 1991 Jan;5(1):49–59. [PubMed]
  • Nash HA, Robertson CA, Flamm E, Weisberg RA, Miller HI. Overproduction of Escherichia coli integration host factor, a protein with nonidentical subunits. J Bacteriol. 1987 Sep;169(9):4124–4127. [PMC free article] [PubMed]
  • Pabo CO, Sauer RT. Protein-DNA recognition. Annu Rev Biochem. 1984;53:293–321. [PubMed]
  • Renna MC, Najimudin N, Winik LR, Zahler SA. Regulation of the Bacillus subtilis alsS, alsD, and alsR genes involved in post-exponential-phase production of acetoin. J Bacteriol. 1993 Jun;175(12):3863–3875. [PMC free article] [PubMed]
  • Rossmann R, Sawers G, Böck A. Mechanism of regulation of the formate-hydrogenlyase pathway by oxygen, nitrate, and pH: definition of the formate regulon. Mol Microbiol. 1991 Nov;5(11):2807–2814. [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]
  • Sauter M, Böhm R, Böck A. Mutational analysis of the operon (hyc) determining hydrogenase 3 formation in Escherichia coli. Mol Microbiol. 1992 Jun;6(11):1523–1532. [PubMed]
  • Sawers G, Böck A. Novel transcriptional control of the pyruvate formate-lyase gene: upstream regulatory sequences and multiple promoters regulate anaerobic expression. J Bacteriol. 1989 May;171(5):2485–2498. [PMC free article] [PubMed]
  • Sawers G, Suppmann B. Anaerobic induction of pyruvate formate-lyase gene expression is mediated by the ArcA and FNR proteins. J Bacteriol. 1992 Jun;174(11):3474–3478. [PMC free article] [PubMed]
  • Schell MA. Molecular biology of the LysR family of transcriptional regulators. Annu Rev Microbiol. 1993;47:597–626. [PubMed]
  • Schlaman HR, Okker RJ, Lugtenberg BJ. Regulation of nodulation gene expression by NodD in rhizobia. J Bacteriol. 1992 Aug;174(16):5177–5182. [PMC free article] [PubMed]
  • Schlensog V, Böck A. Identification and sequence analysis of the gene encoding the transcriptional activator of the formate hydrogenlyase system of Escherichia coli. Mol Microbiol. 1990 Aug;4(8):1319–1327. [PubMed]
  • Simons RW, Houman F, Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. [PubMed]
  • Stormer FC. Evidence for induction of the 2,3-butanediol-forming enzymes in Aerobacter aerogenes. FEBS Lett. 1968 Nov;2(1):36–38. [PubMed]
  • Störmer FC. The pH 6 acetolactate-forming enzyme from Aerobacter aerogenes. I. Kinetic studies. J Biol Chem. 1968 Jul 10;243(13):3735–3739. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Zinoni F, Birkmann A, Stadtman TC, Böck A. Nucleotide sequence and expression of the selenocysteine-containing polypeptide of formate dehydrogenase (formate-hydrogen-lyase-linked) from Escherichia coli. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4650–4654. [PMC free article] [PubMed]

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


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...