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A Bifunctional Endoglucanase/Endoxylanase from Cellulomonas flavigena with Potential Use in Industrial Processes at Different pH

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Abstract

Cellulomonas flavigena CDBB-531 was found to secrete a bifunctional cellulase/xylanase with a molecular mass of 49 kDa and pI 4.3. This enzyme was active on Remazol brilliant blue–carboxymethylcellulose (RBB-CMC) and Remazol brilliant blue–xylan (RBB-X). Based on thin-layer chromatographic analysis of the degradation products, the cellulase activity produced glucose, cellobiose, cellotriose, and cellotetraose from CMC as the substrate. When xylan from birchwood was used, end products were xylose, arabinose, and xylobiose. The bifunctional enzyme showed a pH optimum of 6 for cellulase activity and 9 for xylanase activity, which pointed out that this enzyme had separate sites for each activity. In both cases, the apparent optimum temperature was 50°C. The predicted amino acid sequence of purified protein showed similarity with the catalytic domain of several glycosyl hydrolases of family 10.

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References

  1. Beg QK, Kapoor M, Mahajan L et al (2001) Microbial xylanases and their industrial application: a review. Appl Microbiol Biotechnol 56:326–338

    Article  PubMed  CAS  Google Scholar 

  2. Biely P, Mislovicová D, Toman R (1988) Remazol brilliant blue-xylan: a soluble chromogenic substrate for xylanases. Methods Enzymol 160:536–541

    Article  CAS  Google Scholar 

  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  4. Flint HJ, Martin J, McPherson CA et al (1993) A bifunctional enzyme, with separated xylanase and β (1,3-1,4)-glucanase domains, encoded by the xynD gene of Ruminococcus flavefaciens. J Bacteriol 175:2943–2951

    PubMed  CAS  Google Scholar 

  5. Gilbert HJ, Hazlewood GP (1993) Bacterial cellulases and xylanases. J Gen Microbiol 139:187–194

    CAS  Google Scholar 

  6. Gutierrez-Nava A, Herrera-Herrera A, Mayorga-Reyes L et al (2003) Expression and characterization of the celcflB gene from Cellulomonas flavigena encoding an endo-β-1,4-glucanase. Curr Microbiol 47:359–363

    Article  PubMed  CAS  Google Scholar 

  7. Henrissat B, Bairoch A (1996) Updatind the sequence-based classification of glycosyl hydrolases. Biochem J 316:695–696

    PubMed  Google Scholar 

  8. Kinter M, Sherman NE (2000) Protein sequencing and identification using tandem mass spectrometry. Wiley–Interscience, New York

    Google Scholar 

  9. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  10. MacLeod AM, Lindhoest T, Withers SG et al (1994) The acid/base catalyst in the exoglucanase/xylanase from Cellulomonas fimi is glutamic acid 127: evidence from detailed kinetic studies of mutants. Biochemistry 33:6371–6376

    Article  PubMed  CAS  Google Scholar 

  11. Martínez-Trujillo A, Pérez-Avalos O, Ponce-Noyola T (2003) Enzymatic properties of a purified xylanase from mutant PN-120 of Cellulomonas flavigena. Enzyme Microbiol Technol 32:401–406

    Article  CAS  Google Scholar 

  12. Mayorga-Reyes L, Morales Y, Salgado LM et al (2002) Cellulomonas flavigena: characterization of an endo-1,4-xylanase tightly induced by sugar cane bagasse. FEMS Microbiol Lett 214:205–209

    Article  PubMed  CAS  Google Scholar 

  13. Natesh R, Bhanumoorthy P, Vithayathil PJ et al (1999) Crystal structure at 1.8 Å resolution and proposed amino acid sequence of a thermostable xylanase from Thermoascus aurantiacus. J Mol Biol 288:999–1012

    Article  PubMed  CAS  Google Scholar 

  14. Pérez-Avalos O, Ponce-Noyola T (2002) Synthesis and regulation of d-xylanase from Cellulomonas flavigena wild type and a mutant. Biotechnol Lett 24:813–817

    Article  Google Scholar 

  15. Ponce-Noyola T, de la Torre M (1995) Isolation of a high-specific-growth-rate mutant of Cellulomonas flavigena on sugar cane bagasse. Appl Microbiol Biotechnol 45:709–712

    Article  Google Scholar 

  16. Sambrook JE, Fritsch F, Maniatis T (1989) Molecular cloning: A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  17. White A, Withers SG, Gilkes NR et al (1994) Crystal structure of the catalytic domain of the β-1,4-glycanase Cex from Cellulomonas fimi. Biochemistry 33:12546–12552

    Article  PubMed  CAS  Google Scholar 

  18. Wong KK, Tan LU, Saddler JN (1988) Multiplicity of β-1,4-xylanase in microorganisms: functions and applications. Microbiol Rev 52:305–317

    PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by Consejo Nacional de Ciencia y Tecnología-México (45678-Z). The authors thank the W. M. Keck Biomedical Mass Spectrometry Laboratory UVHS for sequencing the protein and Martha Mercado-Morales for technical assistance.

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Authors and Affiliations

  1. Departamento de Biotecnologia y Bioingenieria, CINVESTAV, Av. IPN # 2508 Col. Zacatenco, C.P. 07000, D.F., Mexico

    Odilia Pérez-Avalos, Leticia M. Sánchez-Herrera & Teresa Ponce-Noyola

  2. Departamento de Bioquimica, CINVESTAV, Av. IPN # 2508 Col. Zacatenco, C.P. 07000, D.F., Mexico

    Luis M. Salgado

Authors
  1. Odilia Pérez-Avalos
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  2. Leticia M. Sánchez-Herrera
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  3. Luis M. Salgado
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  4. Teresa Ponce-Noyola
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Correspondence to Teresa Ponce-Noyola.

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Pérez-Avalos, O., Sánchez-Herrera, L.M., Salgado, L.M. et al. A Bifunctional Endoglucanase/Endoxylanase from Cellulomonas flavigena with Potential Use in Industrial Processes at Different pH. Curr Microbiol 57, 39–44 (2008). https://doi.org/10.1007/s00284-008-9149-1

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  • DOI: https://doi.org/10.1007/s00284-008-9149-1

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