Purification and characterization of two chitinase isoforms from the bulbs of gladiolus (gladiolus gandavensis)

Takeshi Yamagami, Yoichiro Mine, Yoichi Aso, Masatsune Ishiguro

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Abstract

Two chitinase isoforms, designated GBC-a and GBC-b, were purified from the bulbs of gladiolus (Gladiolus gandavensis) using CM-cellulose column chromatography followed by Butyl-Toyopearl 650M hydrophobic column chromatography, gel filtration on Sephadex G-75, and Mono-S FPLC. GBC-a and GBC-b are weakly acidic and weakly basic proteins with molecular masses of 30 kDa, and isoelectric points of 6.0 and 7.5, respectively. GBC-a and GBC-b were found to be homologous proteins with similar amino acid compositions and N-terminal sequences. The number of half-cystine residues in GBC-a and GBC-b was only one each, which is much lower than those of plant class I (15-17 Cys residues/mol), class II (5-8 Cys residues/mol), and class III (6 Cys residues/mol) chitinases. The N-terminal sequences of GBC-a and GBC-b were completely different from those of plant three classes of chitinases. The optimal pHs of these chitinases toward glycolchitin were pH 5. GBC-a hydrolyzed (GlcNAc)6 into (GlcNAc)2, (GlcNAc)3 and (GlcNAc)4, and (GlcNAc)5, into (GlcNAc)2, and (GlcNAc)3.

Original languageEnglish
Pages (from-to)2140-2142
Number of pages3
JournalBioscience, Biotechnology and Biochemistry
Volume61
Issue number12
DOIs
Publication statusPublished - Jan 1997

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All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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