Purification and Antifungal Activity of Recombinant Chitinase fromEscherichia coliCarrying the Family 19 Chitinase Gene ofStreptomycessp. J-13-3
- 1 January 2004
- journal article
- Published by Oxford University Press (OUP) in Bioscience, Biotechnology, and Biochemistry
- Vol. 68 (10) , 2193-2196
Abstract
A recombinant chitinase was purified from the cell extract of Escherichia coli JM109 transformed by plasmid pUC19 carrying the gene encoding family 19 chitinase of Streptomyces sp. J-13-3 by column chromatography on DEAE-Sepharose, CM-Sepharose, and Bio-Gel P-100. The final preparation was homogenous in polyacrylamide gel electrophoresis. The molecular weight of the purified enzyme was estimated to be 32,000. The recombinant chitinase hydrolyzed the trimer to hexamer of N-acetylglucosamine and had the identical N-terminal amino acid sequence of the mature protein, indicating removal of the signal sequence by E. coli signal peptidase. The fungal growth in well (200 microl of medium) of microplate by measurement of absorbance at 595 nm indicated that the chitinase (10 microg) completely and half inhibited growth of Trichoderma reesei and Aspergillus niger respectively.Keywords
This publication has 9 references indexed in Scilit:
- Molecular Cloning and Expression of the Gene Encoding Family 19 Chitinase fromStreptomycessp. J-13-3Bioscience, Biotechnology, and Biochemistry, 2004
- Characterization of Chitinase Genes from an Alkaliphilic Actinomycete, Nocardiopsis prasina OPC-131Applied and Environmental Microbiology, 2003
- Family 19 Chitinase ofStreptomyces griseusHUT6037 Increases Plant Resistance to the Fungal DiseaseBioscience, Biotechnology, and Biochemistry, 2003
- Plant Chitinase as a Possible Biocontrol Agent for Use Instead of Chemical FungicidesBioscience, Biotechnology, and Biochemistry, 2003
- Structure, Heterologous Expression, and Properties of Rice (Oryza sativaL.) Family 19 ChitinasesBioscience, Biotechnology, and Biochemistry, 2003
- Family 19 Chitinases from Streptomyces thermoviolaceus OPC-520: Molecular Cloning and CharacterizationBioscience, Biotechnology, and Biochemistry, 2000
- Family 19 chitinases of Streptomyces species: characterization and distribution The GenBank accession numbers for the sequences determined in this work are AB031745–AB031757 inclusive.Microbiology, 1999
- Purification and Properties of Two Chitinases fromStreptomycessp. J-13-3Bioscience, Biotechnology, and Biochemistry, 1995
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970