Abstract
A multienzyme complex from newly isolated Paenibacillus sp. TW1 was purified from pellet-bound enzyme preparations by elution with 0.25% sucrose and 1.0% triethylamine (TEA), ultrafiltration and Sephacryl S-400 gel filtration chromatography. The purified multienzyme complex showed a single protein band on non-denaturing polyacrylamide gel electrophoresis (native-PAGE). The high molecular mass of the purified multienzyme complex was approximately 1,950 kDa. The complex consisted of xylanase and cellulase activities as the major and minor enzyme subunits, respectively. The complex appeared as at least 18 protein bands on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and as 15 xylanases and 6 cellulases on zymograms. The purified multienzyme complex contained xylanase, α-L-arabinofuranosidase, carboxymethyl cellulase (CMCase), avicelase and cellobiohydrolase. The complex could effectively hydrolyze corn hulls, corncobs and sugarcane bagasse. These results indicate that the multienzyme complex that is produced by this bacterium is a large, novel xylanolytic-cellulolytic enzyme complex.
Article PDF
Similar content being viewed by others
References
Bayer, E. A., Belaich, J. P., Shoham, Y., Lame, R. (2004) The cellulosomes: multienzyme machines for degradation of plant cell wall polysaccharides. Annu. Rev. Microbiol. 58, 521–554.
Bayer, E. A., Chanzy, H., Lamed, R., Shoham, Y. (1998) Cellulose, cellulases and cellulosomes. Curr. Opin. Struct. Biol. 8, 548–557.
Béguin, P., Lemaire, M. (1996) The cellulosome: an exocellular, multiprotein complex specialized in cellulose degradation. Crit. Rev. Biochem. Mol. Biol. 31, 201–236.
Bélaich, J. P., Tardif, C., Bélaich, A., Gaudin, C. (1997) The cellulolytic system of Clostridium cellulolyticum. J. Biotechnol. 57, 3–14.
Berg, B., von Hofsten, B., Pettersson, G. (1972) Growth and cellulase formation by Celluvibrio fulvus. J. Appl. Bacteriol. 35, 201–214.
Coughlan, M. P., Hazlewood, G. P. (1993) ß-1,4-D-Xylan-degrading enzyme system: biochemistry, molecular biology and applications. Biotechnol. Appl. Biochem. 17, 259–289.
Demain, A. L., Newcomb, M., Wu, J. H. D. (2005) Cellulase, clostridia, and ethanol. Microbiol. Mol. Biol. Rev. 69, 124–154.
Doi, R. H., Kosugi, A. (2004) Cellulosomes: plant-cell-wall-degrading enzyme complexes. Nat. Rev. Microbiol. 2, 541–551.
Dyk, J. S. V., Sakka, M., Sakka, K., Pletschke, B. I. (2009) The cellulolytic and hemi-cellulolytic system of Bacillus licheniformis SVD1 and the evidence for production of a large multi-enzyme complex. Enzyme Microb. Technol. 45, 372–378.
Fan, L. T., Gharpuray, M. M., Lee, Y. H. (1987) Cellulose hydrolysis. In: Aiba, S., Fan, L. T., Fiechter, A., Klein, J., Schugerl, K. (eds) Biotechnology Monographs. Springer-Verlag, Berlin Heidelberg, pp. 1–16.
Ghangas, G. S., Hu, Y. J., Wilson, D. B. (1989) Cloning of a Thermomonospora fusca xylanase gene and its expression in Escherichia coli and Streptomyces lividans. J. Bacteriol. 171, 2963–2969.
Irwin, D., Jung, E. D., Wilson, D. B. (1994) Characterization and sequence of a Thermomonospora fusca xylanase. Appl. Environ. Microbiol. 60, 763–770.
Jiang, Z. Q., Deng, W., Li, X. T., Ai, Z. L., Li, L. T., Kusakabe, I. (2005) Characterization of a novel, ultra-large xylanolytic complex (xylanosome) from Streptomyces olivaceoviridis E-86. Enzyme Microb. Technol. 36, 923–929.
Kosugi, A., Murashima, K., Tamura, Y., Doi, R. H. (2002) Cell surface anchoring role of N-terminal surface layer homology domains of Clostridium cellulovorans EngE. J. Bacteriol. 184, 884–888.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Lamed, R., Setter, E., Bayer, E. A. (1983) Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum. J. Bacteriol. 156, 828–836.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275.
Lynd, L. R., Wyman, C. E., Gerngross, T. U. (1999) Biocommodity engineering. Biotechnol. Prog. 15, 777–793.
Nelson, N. (1944) A photometric adaptation of the Somogyi method for the determination of glucose. J. Biol. Chem. 153, 375–380.
Pason, P., Kyu, K. L., Ratanakhanokchai, K. (2006) Paenibacillus curdlanolyticus strain B-6 xylanolytic-cellulolytic enzyme system that degrade insoluble polysaccharides. Appl. Environ. Microbiol. 72, 2483–2490.
Phitsuwan, P., Tachaapaikoon, C., Kosugi, A., Mori, Y., Kyu, K. L., Ratanakhanokchai, K. (2010) A cellulolytic and xylanolytic enzyme complex from an alkalothermoanaerobacterium, Tepidimicrobium xylanilyticum BT14. J. Microbiol. Biotechnol. 20, 893–903.
Ratanakhanokchai, K, Kyu, K. L., Tanticharoen, M. (1999) Purification and properties of a xylanbinding endoxylanase from alkaliphilic Bacillus sp. strain K-1. Appl. Environ. Microbiol. 65, 694–697.
Schwarz, W. H. (2001) The cellulosome and cellulose degradation by anaerobic bacteria. Appl. Microbiol. Biotechnol. 56, 634–649.
Shoham, Y., Lamed, R., Bayer, E. A. (1999) The cellulosome concept as an efficient microbial strategy for the degradation of insoluble polysaccharides. Trends Microbiol. 7, 275–281.
Singleton, P. (2004) Bacteria in Biology, Biotechnology and Medicine, John Wiley and Sons, West Sussex.
Sjostrom, E. (1993) Wood chemistry: fundamentals and applications. Academic Press, San Diego.
Sneath, P. H. A., Mair, N. S., Sharpe, M. E., Holt, J. G. (1986) Bergey’s Manual of Systematic Bacteriology. Williams & Wilkins, Baltimore, Md.
Tachaapaikoon, C., Kyu, K. L., Ratanakhanokchai, K. (2006) Purification of xylanase from alkaliphilic Bacillus sp. K-8 by using corn husk column. Proc. Biochem. 41, 2441–2445.
Tamura, Y., Doi, R. H. (1999) Three surface layer homology domains at the N terminus of the Clostridium cellulovorans major cellulosomal subunit EngE. J. Bacteriol. 181, 3270–3276.
Teeri, T. T. (1997) Crystalline cellulose degradation: new insight into the function of cellobiohydrolases. Trends Biotechnol. 15, 160–167.
Waeonukul, R., Kyu, K. L., Sakka, K., Ratanakhanokchai, K. (2008) Effect of carbon sources on the induction of xylanolytic-cellulolytic multienzyme complexs in Paenibacillus curdlanolyticus strain B-6. Biosci. Biotechnol. Biochem. 72, 321–328.
Waeonukul, R., Kyu, K. L., Sakka, K., Ratanakhanokchai, K. (2009) Isolation and characterization of a multienzyme complex (cellulosome) of the Paenibacillus curdlanolyticus strain B-6 grown on Avicel under aerobic conditions. J. Biosci. Bioeng. 107, 610–614.
Warren, R. A. J. (1996) Microbial hydrolysis of polysaccharides. Annu. Rev. Microbiol. 50, 183–212.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Tachaapaikoon, C., Kyu, K.L., Pason, P. et al. A Novel Multienzyme Complex from a Newly Isolated Facultative Anaerobic Bacterium, Paenibacillus sp. TW1. BIOLOGIA FUTURA 63, 288–300 (2012). https://doi.org/10.1556/ABiol.63.2012.2.10
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1556/ABiol.63.2012.2.10