Gene Cloning, Expression, and Characterization of a Family 51 α-l-Arabinofuranosidase from Streptomyces sp. S9
- 198 Downloads
An α-l-arabinofuranosidase gene, abf51S9, was cloned from Streptomyces sp. S9 and successfully expressed in Escherichia coli BL21 (DE3). The full-length gene consisted of 1,506 bp and encoded 501 amino acids with a calculated mass of 55.2 kDa. The deduced amino acid sequence was highly homologous with the α-l-arabinofuranosidases belonging to family 51 of the glycoside hydrolases. The recombinant protein was purified to electrophoretic homogeneity by Ni-NTA affinity chromatography and subsequently characterized. The optimal pH and temperature for the recombinant enzyme were 6.0 and 60∼65 °C, respectively. The enzyme showed a broad pH range of stability, retaining over 75% of the maximum activity at pH 5.0 to 11.0. The specific activity, K m, and V max with p-nitrophenyl-α-l-arabinofuranoside as substrate were 60.0 U mg−1, 1.45 mM, and 221 μmol min−1 mg−1, respectively. Abf51S9 showed a mild but significant synergistic effect in combination with xylanase on the degradation of oat-spelt xylan and soluble wheat arabinoxylan substrates with a 1.19- and 1.21-fold increase in the amount of reducing sugar released, respectively. These favorable properties make Abf51S9 a good candidate in various industrial applications.
Keywordsα-l-Arabinofuranosidase Streptomyces sp. S9 Gene cloning and expression Synergistic action
This work was supported by the Chinese National High Technology Research and Development Program (863 Program, Grant No. 2007AA100601) and the Chinese Agricultural Microorganism Collection and Share Program (No. 2005DKA21201).
- 1.Prade, R. A. (1996). Biotechnology and Genetic Engineering Reviews, 13, 101–131.Google Scholar
- 3.Coughlan, M. P., & Hazlewood, G. P. (1993). Biotechnology and Applied Biochemistry, 17, 259–289.Google Scholar
- 5.Kebir, H., Dupont, C., & Morosoli, R. (2000). Biochimica et Biophysica Acta, 1491, 177–184.Google Scholar
- 14.Manin, C., Shareek, F., Morosoli, R., & Kluepfel, D. (1994). The Biochemical journal, 302, 443–449.Google Scholar
- 15.Tajana, E., Fiechter, A., & Zimmermann, W. (1992). Applied and Environmental Microbiology, 58, 1447–1450.Google Scholar
- 18.Li, N., Yang, P., Wang, Y., Luo, H., Meng, K., Wu, N., et al. (2008). Journal of Microbiology and Biotechnology, 18, 410–416.Google Scholar
- 24.Vincent, P., Shareck, F., Dupont, C., Morosoli, R., & Kluepfel, D. (1997). The Biochemical Journal, 322, 845–852.Google Scholar