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Pre and post cloning characterization of a β-1,4-endoglucanase from Bacillus sp.

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Abstract

Consistent with its precloning characterization from the cellulolytic Bacillus sp., β-1,4-endoglucanase purified from the recombinant E. coli exhibited maximum activity at 60°C and pH 7.0. It was highly specific for CMC hydrolysis, with stability up to 70°C and over a pH range of 6.0–8.0. The K m and V max values for CMCase activity of the enzyme were 4.1 mg/ml and 25 μmole/ml min−1, respectively. The purified enzyme was a monomer of 65 kDa, as determined by SDS-PAGE. The presence of sucrose and IPTG in fermentation media increased the endoglucanase activity of the recombinant enzyme to 5.2-folds as compared with that of the actual one.

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Authors and Affiliations

  1. Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, 54590, Pakistan

    Sumra Afzal, Mahjabeen Saleem & Mamoona Naz

  2. Sheikh Zayed Fedral Postgraduate Medical Institute, Lahore, 54590, Pakistan

    Riffat Yasmin

  3. Molecular and Cell Biology Laboratory, Department of Physiology and Cell Biology, University of Health Sciences (UHS), Khayaban-e-Jamia Punjab, Lahore, 54600, Pakistan

    Muhammad Imran

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  1. Sumra Afzal
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  2. Mahjabeen Saleem
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  5. Muhammad Imran
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Correspondence to Mahjabeen Saleem.

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Afzal, S., Saleem, M., Yasmin, R. et al. Pre and post cloning characterization of a β-1,4-endoglucanase from Bacillus sp.. Mol Biol Rep 37, 1717–1723 (2010). https://doi.org/10.1007/s11033-009-9592-5

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  • DOI: https://doi.org/10.1007/s11033-009-9592-5

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