Depletion of the xynB2 gene upregulates β-xylosidase expression in C. crescentus

Appl Biochem Biotechnol. 2014 Jan;172(2):1085-97. doi: 10.1007/s12010-013-0549-8. Epub 2013 Oct 19.

Abstract

Caulobacter crescentus is able to express several enzymes involved in the utilization of lignocellulosic biomasses. Five genes, xynB1-5, that encode β-xylosidases are present in the genome of this bacterium. In this study, the xynB2 gene, which encodes β-xylosidase II (CCNA_02442), was cloned under the control of the PxylX promoter to generate the O-xynB2 strain, which overexpresses the enzyme in the presence of xylose. In addition, a null mutant strain, Δ-xynB2, was created by two homologous recombination events where the chromosomal xynB2 gene was replaced by a copy that was disrupted by the spectinomycin-resistant cassette. We demonstrated that C. crescentus cells lacking β-xylosidase II upregulates the xynB genes inducing β-xylosidase activity. Transcriptional analysis revealed that xynB1 (RT-PCR analysis) and xynB2 (lacZ transcription fusion) gene expression was induced in the Δ-xynB2 cells, and high β-xylosidase activity was observed in the presence of different agro-industrial residues in the null mutant strain, a characteristic that can be explored and applied in biotechnological processes. In contrast, overexpression of the xynB2 gene caused downregulation of the expression and activity of the β-xylosidase. For example, the β-xylosidase activity that was obtained in the presence of sugarcane bagasse was 7-fold and 16-fold higher than the activity measured in the C. crescentus parental and O-xynB2 cells, respectively. Our results suggest that β-xylosidase II may have a role in controlling the expression of the xynB1 and xynB2 genes in C. crescentus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Caulobacter crescentus / enzymology*
  • Caulobacter crescentus / genetics*
  • Gene Deletion*
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial*
  • Genetic Complementation Test
  • Industry
  • Mutation / genetics
  • Phenotype
  • Promoter Regions, Genetic / genetics
  • Transcription, Genetic
  • Up-Regulation / genetics*
  • Xylosidases / genetics*
  • Xylosidases / metabolism

Substances

  • Xylosidases
  • exo-1,4-beta-D-xylosidase