Abstract
The metagenomic approach has been used successfully to isolate novel biocatalyst gene from uncultured microorganisms. The gene encoding exo-1,4-β-glucanase avicelase was amplified from the metagenome of the Equus burchelli fecal sample and cloned. The gene was found to be of 1,007 bp of nucleotide which encodes a protein of 318 amino acids with a calculated MW of 36 kDa. The deduced amino acid sequence was homologous with cellulases belonging to the glycosyl hydrolases 6 superfamily. The expressed protein was active towards the substrates avicel and carboxymethyl cellulose, indicating that it has bifunctional cellulolytic enzyme activity. The recombinant protein showed an activity of 5.23 U with specific activity of 6.8 U mg−1 protein with the substrate avicel, while when CMC was used, an activity of 3.0 U with a specific activity of 4.2 U mg−1 protein was achieved. Its optimum pH was determined to be 7.0 and optimum temperature of 35°C.
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The authors gratefully acknowledge the financial support of ICAR for conducting this research under the National Agricultural Innovative project (NAIP).
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Chandrasekharaiah, M., Thulasi, A., Bagath, M. et al. Identification of Cellulase Gene from the Metagenome of Equus burchelli Fecal Samples and Functional Characterization of a Novel Bifunctional Cellulolytic Enzyme. Appl Biochem Biotechnol 167, 132–141 (2012). https://doi.org/10.1007/s12010-012-9660-5
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DOI: https://doi.org/10.1007/s12010-012-9660-5