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Cloning of the Thermostable Cellulase Gene from Newly Isolated Bacillus subtilis and its Expression in Escherichia coli

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Abstract

A bacterial strain with high cellulase activity (0.26 U/ml culture medium) was isolated from hot spring, and classified and named as B. subtilis DR by morphological and 16SrDNA gene sequence analysis. A thermostable endocellulase, CelDR, was purified from the isolated strain. The optimum temperature of the enzyme reaction was 50°C, and CelDR retained 70% of its maximum activity at 75°C after incubation for 30 min. The putative gene celDR, consisting an open reading frame (ORF) of 1,524 nucleotides and encoding a protein of 508 amino acids with a molecular weight of 55 kDa, was purified from B. subtilis DR and cloned into pET-28a for expression. The cellulase production in E. coli BL21 (DE3) was enhanced to approximately three times that of the wild-type strain.

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Acknowledgments

We gratefully acknowledge the financial support of the National New Productions Project from the Science and Technology Ministry (P. R. China).

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Authors and Affiliations

  1. College of Animal Sciences, Northwest A&F University, Yangling, 712100, People’s Republic of China

    Wang Li, Ming-Ming Yang & Yu-Lin Chen

  2. Yangguang-Guangji Medical R&D Co. Ltd., Wuhan, 430062, People’s Republic of China

    Wang Li, Wei-Wei Zhang & Ming-Ming Yang

  3. College of Animal Sciences, Henan S&T University, Luoyang, 471003, People’s Republic of China

    Wang Li

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  1. Wang Li
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Correspondence to Ming-Ming Yang or Yu-Lin Chen.

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Li, W., Zhang, WW., Yang, MM. et al. Cloning of the Thermostable Cellulase Gene from Newly Isolated Bacillus subtilis and its Expression in Escherichia coli . Mol Biotechnol 40, 195–201 (2008). https://doi.org/10.1007/s12033-008-9079-y

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