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KMID : 0613820120220070912
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Journal of Life Science
2012 Volume.22 No. 7 p.912 ~ p.919
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Characterization of Cellulase and Xylanase from Bacillus subtilis NC1 Isolated from Environmental Soil and Determination of Its Genes
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Park Chang-Su
Kang Dae-Ook
Choi Nack-Shick
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Abstract
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A Bacillus sp. strain producing celluase and xylanase was isolated from environmental soil with LB agar plate containing carboxymethylcellulose (CM-cellulose) and beechwood xylan stained with trypan blue as substrates, respectively. Based on the 16S rRNA gene sequence and API 50 CHL test, the strain was identified as B. subtilis and named B. subtilis NC1. The cellulase and xylanase from B. subtilis NC1 exhibited the highest activities for CM-cellulose and beechwood xylan as substrate, respectively, and both enzymes showed the maximum activity at pH 5.0 and 50℃. We cloned and sequenced the genes for cellulase and xylanase from genomic DNA of the B. subtilis NC1 by the shot-gun cloning method. The cloned cellulase and xylanase genes consisted of a 1,500 bp open reading frame (ORF) encoding a 499 amino acid protein with a calculated molecular mass of 55,251 Da and a 1,269 bp ORF encoding a 422 amino acid protein with a calculated molecular mass of 47,423 Da, respectively. The deduced amino acid sequences from the genes of cellulase and xylanase showed high identity with glycosyl hydrolases family (GH) 5 and 30, respectively.
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KEYWORD
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Bacillus subtilis, screening, cellulase, xylanase, characterization, cloning
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