Five extracellular chitinases of Bacillus cereus 6E1 were detected by a novel in-gel chitinase assay using carboxymethyl-chitin-remazol brilliant violet 5R (CM-chitin-RBV) as a substrate. The major chitinase activity was associated with a 36-kDa (Chi36) gel band. Chi36 was purified by a one-step, native gel purification procedure derived from the new in-gel chitinase assay. The purified Chi36 has optimal activity at pH 5.8 and retains some enzymatic activity between pH 2.5-8. The temperature optimum for Chi36 was 35 degrees C, but the enzyme was active between 4-70 degrees C. Based on its ability to hydrolyze mainly p-nitrophenyl-(N-acetyl-beta-D-glucosaminide)(2), Chi36 is characterized as a chitobiosidase, a type of exochitinase. The N-terminal amino acid sequence of mature Chi36 was determined (25 amino acids). Alanine is the first N-terminal amino acid residue indicating the cleavage of a signal peptide from a Chi36 precursor to form the mature extracellular Chi36. The N-terminal sequence of Chi36 demonstrated highest similarity with Bacillus circulans WL-12 chitinase D and significant similarity with several other bacterial chitinases.
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http://dx.doi.org/10.1016/s0141-0229(00)00362-8 | DOI Listing |
3 Biotech
April 2019
1Department of Microbiology, Faculty of Liberal Arts and Science, Kasetsart University, Nakhon Pathom, 73140 Thailand.
A total of 511 local isolates of from different geographical regions of Thailand were analyzed for the presence of the , and genes encoding for lepidopteran-specific toxins. PCR results revealed that 94.32% (482/511) of isolates harbored at least one of the detected genes, of which the , and genes were detected at frequencies of 90.
View Article and Find Full Text PDFJ Biosci Bioeng
November 2005
Department of Biological Sciences, 101 Rouse Life Sciences Building, Auburn University, Auburn, Alabama 36849, USA.
The chi36 gene encoding exochitinase Chi36 was cloned from a Bacillus cereus 6E1 subgenomic library. The chi36 open reading frame is 1080 bp long encoding a Chi36 precursor protein of 360 amino acids, consisting of a 27 amino acid N-terminal signal peptide and a 333 amino acid sequence found in the mature Chi36 protein of 36.346 kDa.
View Article and Find Full Text PDFBiochem Biophys Res Commun
August 2003
International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, PO Box 10504, New Delhi 1100 67, India.
A 36 kDa chitinase was purified by ion exchange and gel filtration chromatography from the culture supernatant of Bacillus thuringiensis HD-1. The chitinase production was independent of the presence of chitin in the growth medium and was produced even in the presence of glucose. The purified chitinase was active at acidic pH, had an optimal activity at pH 6.
View Article and Find Full Text PDFEnzyme Microb Technol
April 2001
Department of Biological Sciences, Auburn University, 101 Life Sciences Building, 36849, Auburn, AL, USA
Five extracellular chitinases of Bacillus cereus 6E1 were detected by a novel in-gel chitinase assay using carboxymethyl-chitin-remazol brilliant violet 5R (CM-chitin-RBV) as a substrate. The major chitinase activity was associated with a 36-kDa (Chi36) gel band. Chi36 was purified by a one-step, native gel purification procedure derived from the new in-gel chitinase assay.
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