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Journal of Bacteriology, January 1999, p. 319-330, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Regulation of the sol Locus Genes for Butanol and Acetone Formation in Clostridium acetobutylicum ATCC 824 by a Putative Transcriptional Repressor

Ramesh V. Nair,1,dagger Edward M. Green,2 David E. Watson,2 George N. Bennett,2 and Eleftherios T. Papoutsakis1,*

Department of Chemical Engineering, Northwestern University, Evanston, Illinois 60208,1 and Department of Biochemistry and Cell Biology, Rice University, Houston, Texas 772512

Received 4 June 1998/Accepted 28 October 1998

A gene (orf1, now designated solR) previously identified upstream of the aldehyde/alcohol dehydrogenase gene aad (R. V. Nair, G. N. Bennett, and E. T. Papoutsakis, J. Bacteriol. 176:871-885, 1994) was found to encode a repressor of the sol locus (aad, ctfA, ctfB and adc) genes for butanol and acetone formation in Clostridium acetobutylicum ATCC 824. Primer extension analysis identified a transcriptional start site 35 bp upstream of the solR start codon. Amino acid comparisons of SolR identified a potential helix-turn-helix DNA-binding motif in the C-terminal half towards the center of the protein, suggesting a regulatory role. Overexpression of SolR in strain ATCC 824(pCO1) resulted in a solvent-negative phenotype owing to its deleterious effect on the transcription of the sol locus genes. Inactivation of solR in C. acetobutylicum via homologous recombination yielded mutants B and H (ATCC 824 solR::pO1X) which exhibited deregulated solvent production characterized by increased flux towards butanol and acetone formation, earlier induction of aad, lower overall acid production, markedly improved yields of solvents on glucose, a prolonged solvent production phase, and increased biomass accumulation compared to those of the wild-type strain.

* Corresponding author. Mailing address: Department of Chemical Engineering, Northwestern University, 2145 Sheridan Rd., Evanston, IL 60208-3120. Phone: (847) 491-7455. Fax: (847) 491-3728. E-mail: e-paps{at}nwu.edu.

dagger Present address: DuPont Life Sciences, Experimental Station, Wilmington, DE 19880-0328.

Journal of Bacteriology, January 1999, p. 319-330, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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