title: ICChI, a glycosylated chitinase from the latex of Ipomoea carnea.
abstract: A multi-functional enzyme ICChI with chitinase/lysozyme/exochitinase activity from the latex of Ipomoea carnea subsp. fistulosa was purified to homogeneity using ammonium sulphate precipitation, hydrophobic interaction and size exclusion chromatography. The enzyme is glycosylated (14-15%), has a molecular mass of 34.94 kDa (MALDI-TOF) and an isoelectric point of pH 5.3. The enzyme is stable in pH range 5.0-9.0, 80 degrees C and the optimal activity is observed at pH 6.0 and 60 degrees C. Using p-nitrophenyl-N-acetyl-beta-D-glucosaminide, the kinetic parameters K(m), V(max), K(cat) and specificity constant of the enzyme were calculated as 0.5mM, 2.5 x 10(-8)mol min(-1)microg enzyme(-1), 29.0 s(-1) and 58.0mM(-1)s(-1) respectively. The extinction coefficient was estimated as 20.56 M(-1)cm(-1). The protein contains eight tryptophan, 20 tyrosine and six cysteine residues forming three disulfide bridges. The polyclonal antibodies raised and immunodiffusion suggests that the antigenic determinants of ICChI are unique. The first fifteen N-terminal residues G-E-I-A-I-Y-W-G-Q-N-G-G-E-G-S exhibited considerable similarity to other known chitinases. Owing to these unique properties the reported enzyme would find applications in agricultural, pharmaceutical, biomedical and biotechnological fields.
authors.affiliation: Molecular Biology Unit, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Ashok Kumar
authors.initials: AK
authors.lastname: Patel
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Singh
authors.forename: Ravi Prakash
authors.initials: RP
authors.lastname: Yadav
authors.forename: A J G
authors.initials: AJ
authors.lastname: Moir
authors.forename: Medicherla V
authors.initials: MV
authors.lastname: Jagannadham
mesh_terms: D000595:Amino Acid Sequence; D002688:Chitinases / Q000737:chemistry* / Q000378:metabolism*; D006031:Glycosylation; D027741:Ipomoea / Q000201:enzymology*; D007526:Isoelectric Point; D007700:Kinetics; D007840:Latex / Q000737:chemistry*; D008969:Molecular Sequence Data; D013379:Substrate Specificity
chemical_list: D007840:Latex; D002688:Chitinases