Cloning of the Thermostable Cellulase Gene from Newly Isolated Bacillus subtilis and its Expression in Escherichia coli |
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Authors: | Wang Li Wei-Wei Zhang Ming-Ming Yang Yu-Lin Chen |
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Institution: | (1) College of Animal Sciences, Northwest A&F University, Yangling, 712100, People’s Republic of China;(2) Yangguang-Guangji Medical R&D Co. Ltd., Wuhan, 430062, People’s Republic of China;(3) College of Animal Sciences, Henan S&T University, Luoyang, 471003, People’s Republic of China |
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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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Keywords: | Cellulase Heat-resistance Bacillus subtilis Clone and expression |
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