Abstract
Strains of thermophilic bacilli were screened for cellulolytic activity by gel diffusion assay on selective medium at 55°C. Strain B-41361, identified as a strain of Bacillus licheniformis, displayed activity against carboxymethylcellulose. Zymogram analysis demonstrated several catalytically active polypeptides with the most prominent species having a mass of 37 kDa. The enzyme was purified 60-fold with a 17% yield and specific activity of 183 U/mg. The amino terminal sequence was homologous to members of glycoside hydrolase family 5. Optimal temperature was 65°C (measured over 30 min), but the enzyme was most stable at 60°C, retaining greater than 90% activity after one hour. The enzyme had a broad pH range, with maximal activity at pH 6.0, 75% maximal activity at pH 4.5, and 40% at pH 10. The enzyme hydrolyzed p-nitrophenylcellobioside, barley β-glucan, and lichenan, but no activity was detected against avicel or acid-swollen cellulose.
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References
Atlas RM (1993) Bacillus stearothermophilus broth. In: Parks LC (ed) Handbook of Microbiological Media, CRC Press, Boca Raton, Florida, p 109
Bischoff KM, Shi L, Kennelly PJ (1998) The detection of enzyme activity following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Anal Biochem 260:1–17
Bounias M (1980) N-(1-Napthyl)ethylenediamine dihydrochloride as a new reagent for nanamole quantification of sugars on thin-layer plates by a mathematical calibration process. Anal Biochem 106:291–295
Carrard G, Koivula A, Soderlund H, Beguin P (2000) Cellulose-binding domains promote hydrolysis of different sites on crystalline cellulose. Proc Natl Acad Sci USA 97:10342–10347
Liu Y, Zhang J, Liu Q, Zhang C, Ma Q (2004) Molecular cloning of novel cellulase genes cel9A and cel12A from Bacillus licheniformis GXN151 and synergism of their encoded polypeptides. Curr Microbiol 49:234–238
Lynd LR, Weimer PJ, van Zyl WH, Pretorius IS (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66:506–577
Saha BC (2004) Production, purification and properties of endoglucanase from a newly isolated strain of Mucor circinelloides. Process Biochem 39:1871–1876
Stutzenberger FJ (1991) Cellulose degradation by thermophilic aerobic bacteria. In: Haigler CH, Weimer PJ (eds) Biosynthesis and Biodegradation of Cellulose. Marcel Dekker, Inc. New York
Wood PJ, Erfle JD, Teather RM (1988) Use of complex formation between Congo Red and polysaccharides in detection and assay of polysaccharide hydrolases. Methods Enzymol 160:59–74
Wood TM, Bhat KM (1988) Methods for measuring cellulase activities. Methods Enzymol 160:87–112
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The authors wish to thank Eric Hoecker, Theresa Holly, Imran Khan, and Suzanne Platt for technical assistance.
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Bischoff, K.M., Rooney, A.P., Li, XL. et al. Purification and characterization of a family 5 endoglucanase from a moderately thermophilic strain of Bacillus licheniformis . Biotechnol Lett 28, 1761–1765 (2006). https://doi.org/10.1007/s10529-006-9153-0
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DOI: https://doi.org/10.1007/s10529-006-9153-0