Molecular Biotechnology

, Volume 40, Issue 2, pp 195–201 | Cite as

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

  • Wang Li
  • Wei-Wei Zhang
  • Ming-Ming YangEmail author
  • Yu-Lin ChenEmail author


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.


Cellulase Heat-resistance Bacillus subtilis Clone and expression 



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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Copyright information

© Humana Press 2008

Authors and Affiliations

  1. 1.College of Animal SciencesNorthwest A&F UniversityYanglingPeople’s Republic of China
  2. 2.Yangguang-Guangji Medical R&D Co. Ltd.WuhanPeople’s Republic of China
  3. 3.College of Animal SciencesHenan S&T UniversityLuoyangPeople’s Republic of China

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