Publication: Purification and characterization of chitinase from Bacillus circulans No.4.1
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Issued Date
1999-08-24
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ISSN
03438651
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2-s2.0-0032858156
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Mahidol University
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SCOPUS
Bibliographic Citation
Current Microbiology. Vol.39, No.3 (1999), 134-140
Suggested Citation
Chanpen Wiwat, Patcharaporn Siwayaprahm, Amarat Bhumiratana Purification and characterization of chitinase from Bacillus circulans No.4.1. Current Microbiology. Vol.39, No.3 (1999), 134-140. doi:10.1007/s002849900434 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/25432
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Title
Purification and characterization of chitinase from Bacillus circulans No.4.1
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Abstract
Bacillus circulans No.4.1 produced a high level of chitinase when cells were grown in tryptic soy broth supplemented with 0.3% colloidal chitin at 35°C for 5 days. Purification was carried out by protein precipitation with 80% saturation ammonium sulfate, anion-exchange chromatography with DEAE- Sephacel, and gel filtration with Sephadex G-100, sequentially. The purified enzyme could be demonstrated as a single band on SDS-PAGE, estimated to be 45 kDa. This enzyme could hydrolyze colloidal chitin, purified chitin, glycol chitin, carboxymethyl-chitin (CM-chitin), and 4-methylumbelliferyl-β-D- N,N'-diacetylchitobioside [4-MU-(GlcNAc)2]. The optimal conditions for this chitinase were pH 8.0 and 40°C. The isoelectric point of the chitinase was 5.1. The amino acid composition of the purified chitinase was determined. The initial 20 amino acid residues of the N-terminal were found to be alanine (A), proline (P), tryptophan (W), asparagine (N), serine (S), lysine (K), glycine (G), asparagine (N), tyrosine (Y), alanine (A), leucine (L), proline (P), tyrosine (Y), tyrosine (Y), arginine (R), glycine (G), alanine (A), tryptophan (W), alanine (A), and valine (V). Knowledge of these properties of chitinase from B. circulans No. 4.1 should be useful in the development of genetically engineered Bacillus sp. as biopesticides.