Abstract
A cellulase gene (cel28a) was isolated from a rumen microbial metagenome library of goat rumen microorganisms, cloned into E. coli, and expressed in active form. The gene has a length of 1596 bp obtained using a genome walking Kit and encodes a protein of 509 amino acids with a calculated MW of 55 kDa. The deduced amino acid sequence was homologous with cellulases belonging to the glycosyl hydrolase family 5 (GH5). The expressed protein showed activity toward carboxymethylcellulose (CMC) and xylan, suggesting non-specific endoglucanase activity. The optimal conditions for endoglucanase and xylanase activities were 50 °C and pH 5.0. The metal ions (Ca2+, Fe2+, Mn2+ and Co2+) stimulated the cellulase activity of cel28a, while the other metal ions and chemicals (Ni2+, Mg2+, Zn2+, Cu2+, SDS and EDTA) inhibited the cellulase activity. Further examination of substrate preference showed a higher activity with CMC, oat spelt xylan and birchwood xylan than with filter paper and microcrystalline cellulose, again suggesting that the protein was an endoglucanase with xylanase activity.
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This work was supported by the Education Department Natural Science Foundation of Anhui Province (KJ2013A115), Natural Science Foundation of Anhui Province (No. 090411019) and Specialized Research Fund for the Doctoral Program of Higher Education (20133418120002).
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Cheng, J., Huang, S., Jiang, H. et al. Isolation and characterization of a non-specific endoglucanase from a metagenomic library of goat rumen. World J Microbiol Biotechnol 32, 12 (2016). https://doi.org/10.1007/s11274-015-1957-4
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DOI: https://doi.org/10.1007/s11274-015-1957-4