Identification of Cellulase Gene from the Metagenome of Equus burchelli Fecal Samples and Functional Characterization of a Novel Bifunctional Cellulolytic Enzyme

被引:0
作者
Matam Chandrasekharaiah
Appoothy Thulasi
Madiajagan Bagath
Duvvuri Prasanna Kumar
Sunil Singh Santosh
Chenniappan Palanivel
Vazhakkala Lyju Jose
Koratokare Thirumalachar Sampath
机构
[1] National Institute of Animal Nutrition and Physiology,Rumen Microbiology Laboratory, Animal Nutrition Division
来源
Applied Biochemistry and Biotechnology | 2012年 / 167卷
关键词
Exo-1,4-β-glucanase; Avicelase; Metagenome; Enzyme activity; Cloning;
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中图分类号
学科分类号
摘要
The metagenomic approach has been used successfully to isolate novel biocatalyst gene from uncultured microorganisms. The gene encoding exo-1,4-β-glucanase avicelase was amplified from the metagenome of the Equus burchelli fecal sample and cloned. The gene was found to be of 1,007 bp of nucleotide which encodes a protein of 318 amino acids with a calculated MW of 36 kDa. The deduced amino acid sequence was homologous with cellulases belonging to the glycosyl hydrolases 6 superfamily. The expressed protein was active towards the substrates avicel and carboxymethyl cellulose, indicating that it has bifunctional cellulolytic enzyme activity. The recombinant protein showed an activity of 5.23 U with specific activity of 6.8 U mg−1 protein with the substrate avicel, while when CMC was used, an activity of 3.0 U with a specific activity of 4.2 U mg−1 protein was achieved. Its optimum pH was determined to be 7.0 and optimum temperature of 35°C.
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页码:132 / 141
页数:9
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