Abstract
Cellulases degrade cellulose, which is the most abundant biological polymer on the earth (1). Although the chemical composition of cellulose is very simple, consisting of only glucose residues connected by β-1,4-glycosidic bonds, no single enzyme is able to degrade crystalline cellulose. To degrade crystalline cellulose to glucose, at least three enzymes have to cooperate: endoglucanase (EC 3.2.1.4), exoglucanase (cellobiohydrolase, EC 3.2.1.91), and β-glucosidase (EC 3.2.1.21) (1).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Notes
- 1.
*
Sterilize before use
- 2.
*
Sterilize before use
- 3.
*
Sterilize before use
- 4.
*
Sterilize before use
References
Schwarz, W. H. (2001) The cellulosome and cellulose degradation by anaerobic bacteria. Appl. Microbiol. Biotechnol. 56, 634–649.
Lynd, L. R., Cushman, J. H., Nichols, R. J., and Wyman, C. E. (1991) Fuel ethanol from cellulosic biomass. Science 251, 1318–1323.
Azevedo, H., Bishop, D., and Cavaco-Paulo, A. (2000) Effects of agitation level on the adsorption, desorption, and activities on cotton fabrics of full length and core domains of EGV (Humicola insolens) and CenA (Cellulomonas fimi). Enzyme Microb. Technol. 27, 325–329.
Ito, S. (1997) Alkaline cellulases from alkaliphilic Bacillus: enzymatic properties, genetics, and application to detergents. Extremophiles 1, 61–66.
Doi, R. H., Park, J. S., Liu, C. C., et al. (1998) Cellulosome and noncellulosomal cellulases of Clostridium cellulovorans. Extremophiles 2, 53–60.
Shoseyov, O. and Doi, R. H. (1990) Essential 170-kDa subunit for degradation of crystalline cellulose by Clostridium cellulovorans cellulase. Proc. Natl. Acad. Sci. USA 87, 2192–2195.
Tamaru, Y., Karita, S., Ibrahim, A., Chan, H., and Doi, R. H. (2000) A large gene cluster for the Clostridium cellulovorans cellulosome. J. Bacteriol. 182, 5906–5910.
Wood, T. M. and Bhat, K. M. (1988) Methods for measuring cellulase activities, in Methods Enzymology (Wood, W. A. and Kellogg, S. T., eds.), Academic Press, San Diego, CA, pp. 87–112.
Wood, P. J., Erfle, J. D., and Teather, R. M. (1988) Use of complex formation between Congo Red and polysaccharides in detection and assay of polysaccharide hydrolases, in Methods Enzymology (Wood, W. A. and Kellogg, S. T., eds.), Academic Press, San Diego, pp. 59–74.
Murashima, K., Kosugi, A., and Doi, R. H. (2002) Thermostabilization of cellulosomal endoglucanase EngB from Clostridium cellulovorans by in vitro DNA recombination with non-cellulosomal endoglucanase EngD. Mol. Microbiol. 45, 617–626.
Lin, Z., Thorsen, T., and Arnold, F. H. (1999) Functional expression of horseradish peroxidase in E. coli by directed evolution. Biotechnol. Prog. 15, 467–471.
Lavallie, E. R., Lu, Z., Diblasio-Smith, E. A., Collins-Racie, L. A., and Mccoy, J. M. (2000) Thioredoxin as a fusion partner for production of soluble recombinant proteins in Escherichia coli. Methods Enzymol. 326, 322–340.
Hamamoto, T., Foong, F. C., Shoseyov, O., and Doi, R. H. (1992) Analysis of functional domains of endoglucanases from Clostridium cellulovorans by gene cloning, nucleotide sequencing and chimeric protein construction. Mol. Gen. Genet. 231, 472–479.
Zhao, H., Giver, L., Shao, Z., Affholter, J. A., and Arnold, F.,H. (1998) Molecular evolution by staggered extension process (StEP) in vitro recombination. Nat. Biotechnol. 16, 258–261.
Shoichet, B. K., Baase, W. A., Kuroki, R., and Matthews, B. W. (1995) A relationship between protein stability and protein function. Proc. Natl. Acad. Sci. USA 92, 452–456.
Murashima, K., Kosugi, A., and Doi, R. H. (2002) Synergistic effects on crystalline cellulose degradation between cellulosomal cellulases from Clostridium cellulovorans. J. Bacteriol. 184, 5088–5095.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Murashima, K., Doi, R.H. (2003). Selection of Heat-Stable Clostridium cellulovorans Cellulases After In Vitro Recombination. In: Arnold, F.H., Georgiou, G. (eds) Directed Enzyme Evolution. Methods in Molecular Biology™, vol 230. Humana Press. https://doi.org/10.1385/1-59259-396-8:231
Download citation
DOI: https://doi.org/10.1385/1-59259-396-8:231
Publisher Name: Humana Press
Print ISBN: 978-1-58829-286-5
Online ISBN: 978-1-59259-396-5
eBook Packages: Springer Protocols