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Purification and characterization of an organic-solvent-tolerant cellulase from a halotolerant isolate, Bacillus sp. L1

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A halotolerant isolate Bacillus sp. L1 producing extracellular cellulase was isolated from Yuncheng, China. Production of the enzyme started from mid-exponential phase of bacterial growth and reached a maximum level during the post-stationary phase. The cellulase was purified to homogeneity with molecular mass of 45 kDa. Substrate specificity test indicated that it was an endoglucanase for soluble cellulose. Optimal enzyme activity was found to be at 60 °C, pH 8.0, and 7.5 % NaCl. Furthermore, it was highly active and stable over broad ranges of temperature (30–80 °C), pH (7.0–9.0), and NaCl concentration (2.5–15 %), thus showing its excellent thermostable, alkali-stable, and halotolerant nature. The cellulase activity was greatly inhibited by ethylenediaminetetraacetic acid, indicating that it was a metalloenzyme. Significant inhibition by phenylmethylsulfonyl fluoride and phenylarsine oxide revealed that serine and cysteine residues were essential for the enzyme catalysis. Moreover, the cellulase was highly active in the presence of surfactants, and it showed high stability in the presence of water-insoluble organic solvents with log P owat least 0.88. Results from this study indicate that the purified cellulase from isolate L1 may have considerable potential for industrial application owing to its useful properties.

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Acknowledgments

This work was financially supported by Shanxi Provincial Science and Technology Foundation (grant no. 20110021) and Natural Science Fund of Shanxi Province (grant no. 2011021031-4).

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  1. Life Science College, Yuncheng University, Yuncheng, 044000, China

    Xin Li & Hui-Ying Yu

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  1. Xin Li
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  2. Hui-Ying Yu
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Correspondence to Xin Li.

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Li, X., Yu, HY. Purification and characterization of an organic-solvent-tolerant cellulase from a halotolerant isolate, Bacillus sp. L1. J Ind Microbiol Biotechnol 39, 1117–1124 (2012). https://doi.org/10.1007/s10295-012-1120-2

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  • DOI: https://doi.org/10.1007/s10295-012-1120-2

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