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Depletion of the xynB2 Gene Upregulates β-Xylosidase Expression in C. crescentus

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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.

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Acknowledgments

This work was supported by grants from the Araucaria Foundation and CNPq. R.C.G. Simão was partially supported by Araucaria Foundation (process 893/2012). M. R. Mingori and J. M. Corrêa are fellows of the CNPq and Coordination of Improvement of Higher Education Personnel (CAPES), respectively.

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Authors and Affiliations

  1. Centro de Ciências Exatas e Tecnológicas, Universidade Estadual do Oeste do Paraná, Cascavel, Brazil

    Juliana Moço Corrêa

  2. Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Rua Universitária, 2069, Cascavel, Paraná, 85814-110, Brazil

    Moara Rodrigues Mingori & Rita de Cássia Garcia Simão

  3. Hospital Universitário do Oeste do Paraná, Universidade Estadual do Oeste do Paraná, Cascavel, Brazil

    Rinaldo Ferreira Gandra & Eduardo Alexandre Loth

  4. Departamento de Bioquímica, Universidade Estadual de Maringá, Maringá, Paraná, Brazil

    Flávio Augusto Vicente Seixas

Authors
  1. Juliana Moço Corrêa
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  2. Moara Rodrigues Mingori
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  3. Rinaldo Ferreira Gandra
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  4. Eduardo Alexandre Loth
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  5. Flávio Augusto Vicente Seixas
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  6. Rita de Cássia Garcia Simão
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Correspondence to Rita de Cássia Garcia Simão.

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Corrêa, J.M., Mingori, M.R., Gandra, R.F. et al. Depletion of the xynB2 Gene Upregulates β-Xylosidase Expression in C. crescentus . Appl Biochem Biotechnol 172, 1085–1097 (2014). https://doi.org/10.1007/s12010-013-0549-8

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