Abstract
The diversity of cellulases and xylanases secreted by Cellulomonas flavigena cultured on sugar cane bagasse, Solka-floc, xylan, or glucose was explored by two-dimensional gel electrophoresis. C. flavigena produced the largest variety of cellulases and xylanases on sugar cane bagasse. Multiple extracellular proteins were expressed with these growth substrates, and a limited set of them coincided in all substrates. Thirteen proteins with carboxymethyl cellulase or xylanase activity were liquid chromatography/mass spectrometry sequenced. Proteins SP4 and SP18 were identified as products of celA and celB genes, respectively, while SP20 and SP33 were isoforms of the bifunctional cellulase/xylanase Cxo recently sequenced and characterized in C. flavigena. The rest of the detected proteins were unknown enzymes with either carboxymethyl cellulase or xylanase activities. All proteins aligned with glycosyl hydrolases listed in National Center for Biotechnology Information database, mainly with cellulase and xylanase enzymes. One of these unknown enzymes, protein SP6, was cross-induced by sugar cane bagasse, Solka-floc, and xylan. The differences in the expression maps of the presently induced cultures revealed that C. flavigena produces and secretes multiple enzymes to use a wide range of lignocellulosic substrates as carbon sources. The expression of these proteins depends on the nature of the cellulosic substrate.
Similar content being viewed by others
References
Avitia CI, Castellanos-Juárez FX, Sánchez E, Téllez-Valencia A, Fajardo-Cavazos P, Nicholson WL, Pedraza-Reyes M (2000) Temporal secretion of multicellulolytic system in Myxobacter sp. AL-1. Molecular cloning and heterologous expression of cel9 encoding a modular endocellulase clustered in an operon wich cel 48, an exocellobiohydrolase gene. Eur J Biochem 267:7058–7064
Demain AL, Newcomb M, Wu JHD (2005) Cellulase, clostridia, and ethanol. Microbiol Mol Biol Rev 69:124–154
Foreman KP, Brown D, Dankmeyer L, Dean R, Diener S, Dunn-Coleman NS, Goedegebuur F, Houfek TD, England GJ, Kelley AS, Meerman HJ, Mitchell T, Mitchinson C, Olivares HA, Teunissen PJM, Yao J, Ward M (2003) Trascriptional regulation of biomass-degrading enzymes in the filamentous fungus Trichoderma reesei. J Biol Chem 278:31988–31997
Gutiérrez-Nava A, Herrera-Herrera A, Mayorga-Reyes L, Salgado L, Ponce-Noyola T (2003) Characterization and expression of the celcflB gene from Cellulomonas flavigena encoding an endo-β-1,4-glucanase. Curr Microbiol 47:359–363
Han SO, Cho HY, Yukawa H, Inui M, Doi RH (2004) Regulation of expression of cellulosomes and noncellulosomal (hemi)cellulolytic enzymes in Clostridium cellulovorans during growth on different carbon sources. J Bacteriol 186:4218–4227
Kim BH, Wimpenny WT (1981) Grown and cellulolytic activity of Cellulomonas flavigena. Can J Microbiol 27:1260–1266
Lynd LR, Weimer PJ, van Zyl WH (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66:506–577
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
Nochur SV, Roberts MF, Demain AL (1993) True cellulase production by Clostridium thermocellum grown on different carbon sources. Biotechnol Lett 15:641–646
Pérez-Avalos O, Ponce-Noyola T, Magaña I, de la Torre M (1996) Induction of xylanase and β-xylosidase in Cellulomonas flavigena growing on different carbon sources. Appl Microbiol Biotechnol 46:405–409
Pérez-Avalos O, Sánchez-Herrera L, Salgado L, Ponce-Noyola T (2007) A bifunctional endo- glucanase/xylanase from Cellulomonas flavigena with potential use in industrial processes at different pH. Curr Microbiol (in press)
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 42:709–712
Ponce-Noyola T, de la Torre M (2001) Regulation of cellulases and xylanases from a derepressed mutant of Cellulomonas flavigena growing on sugar-cane bagasse in continuous culture. Biores Technol 78:285–91
Rajoka MI (2005) Regulation of synthesis of endo-xylanase and β-xylosidase in Cellulomonas flavigena: a Kinetic study. World J Microbiol Biotechnol 21:463–469
Spiridonov NA, Wilson DB (1988) Regulation of biosynthesis of individual cellulases in Thermomonospora fusca. J Bacteriol 180:3529–3532
Stals I, Sandra K, Devreese B, Van Beeumen J, Claeyssens M (2004) Factors influencing glycosylation of Trichoderma reesei cellulases. II. N-Glycosylation of Cel7A core protein isolated from different strains. Glycobiology 14:725–737
Subramaniyan S, Prema P (2002) Biotechnology of microbial xylanases: enzymology, molecular biology, and application. Crit Rev Biotechnol 22:33–64
Teather RM, Wood PJ (1982) Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from bovine rumen. Appl Environ Microbiol 43:777–780
Wong KY, Tan UL, Saddler NJ (1988) Multiplicity of β-1,4-xylanase in microorganisms: functions and applications. J Microbiol Rev 52:305–317
Acknowledgments
This work was subsidized by Project 45678-Z (CONACYT-México) to T. Ponce-Noyola. L.M. Sánchez-Herrera was supported by PROMEP and Universidad Autónoma de Nayarit, México.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sánchez-Herrera, L.M., Ramos-Valdivia, A.C., de la Torre, M. et al. Differential expression of cellulases and xylanases by Cellulomonas flavigena grown on different carbon sources. Appl Microbiol Biotechnol 77, 589–595 (2007). https://doi.org/10.1007/s00253-007-1190-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-007-1190-7