Mutations that alter the regulation of the chb operon of Escherichia coli allow utilization of cellobiose

Kachroo, Aashiq H and Kancherla, Aswani K and Singh, Nongmaithem S and Varshney, Umesh and Mahadevan, Subramony (2007) Mutations that alter the regulation of the chb operon of Escherichia coli allow utilization of cellobiose. In: Molecular Microbiology, 66 (6). pp. 1382-1395.

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Abstract

Wild-type strains of Escherichia coli are normally unable to metabolize cellobiose. However, cellobiosepositive $(Cel^+)$ mutants arise upon prolonged incubation on media containing cellobiose as the sole carbon source. We show that the $Cel^+$ derivatives carry two classes of mutations that act concertedly to alter the regulation of the chb operon involved in the utilization of $N,N^{\prime}$-diacetylchitobiose. These consist of mutations that abrogate negative regulation by the repressor NagC as well as single base-pair changes in the transcriptional regulator chbR that translate into singleamino-acid substitutions. Introduction of chbR from two $Cel^+$ mutants resulted in activation of transcription from the chb promoter at a higher level in the presence of cellobiose, in reporter strains carrying disruptions of the chromosomal chbR and nagC. These transformants also showed a $Cel^+$ phenotype on MacConkey cellobiose medium, suggesting that the wild-type permease and phospho-\beta-glucosidase, upon induction, could recognize, transport and cleave cellobiose respectively. This was confirmed by expressing the wild-type genes encoding the permease and phospho-\beta-glucosidase under a heterologous promoter. Biochemical characterization of one of the chbR mutants, chbRN238S, showed that the mutant regulator makes stronger contact with the target DNA sequence within the chb promoter and has enhanced recognition of cellobiose 6-phosphate as an inducer compared with the wild-type regulator.

Item Type:	Journal Article
Publication:	Molecular Microbiology
Publisher:	John Wiley & Sons
Additional Information:	Copyright of this article belongs to John Wiley & Sons.
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit Division of Biological Sciences > Molecular Reproduction, Development & Genetics Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited:	16 Sep 2008 09:15
Last Modified:	19 Sep 2010 04:49
URI:	http://eprints.iisc.ac.in/id/eprint/15719

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