title: Purification and characterization of a novel chitinase from Trichosanthes dioica seed with antifungal activity.
abstract: Chitinases are a group of enzymes that show differences in their molecular structure, substrate specificity, and catalytic mechanism and widely found in organisms like bacteria, yeasts, fungi, arthropods actinomycetes, plants and humans. A novel chitinase enzyme (designated as TDSC) was purified from Trichosanthes dioica seed with a molecular mass of 39±1 kDa in the presence and absence of β-mercaptoethanol. The enzyme was a glycoprotein in nature containing 8% neutral sugar. The N-terminal sequence was determined to be EINGGGA which did not match with other proteins. Amino acid analysis performed by LC-MS revealed that the protein was rich in leucine. The enzyme was stable at a wide range of pH (5.0-11.0) and temperature (30-90 °C). Chitinase activity was little bit inhibited in the presence of chelating agent EDTA (ethylenediaminetetraaceticacid), urea and Ca(2+). A strong fluorescence quenching effect was found when dithiothreitol and sodium dodecyl sulfate were added to the enzyme. TDSC showed antifungal activity against Aspergillus niger and Trichoderma sp. as tested by MTT assay and disc diffusion method.
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh. Electronic address: rashelkabir@ru.ac.bd.
authors.forename: Syed Rashel
authors.initials: SR
authors.lastname: Kabir
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Md Musfikur
authors.initials: MM
authors.lastname: Rahman
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Shahnima
authors.initials: S
authors.lastname: Tasnim
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Md Rezaul
authors.initials: MR
authors.lastname: Karim
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Nazma
authors.initials: N
authors.lastname: Khatun
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh; Laboratory of Glycobiology and Marine Biochemistry, Department of Life and Environmental System Science, Graduate School of NanoBio Sciences, Yokohama City University, Yokohama 236-0027, Japan.
authors.forename: Imtiaj
authors.initials: I
authors.lastname: Hasan
authors.affiliation: Bangladesh Council of Scientific and Industrial Research Laboratories, Binodpur Bazar, Rajshahi, Bangladesh.
authors.forename: Ruhul
authors.initials: R
authors.lastname: Amin
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Shaikh Shohidul
authors.initials: SS
authors.lastname: Islam
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Md
authors.initials: M
authors.lastname: Nurujjaman
authors.affiliation: Department of Botany, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Ahmad Humayan
authors.initials: AH
authors.lastname: Kabir
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: Niranjan Kumar
authors.initials: NK
authors.lastname: Sana
authors.affiliation: Laboratory of Glycobiology and Marine Biochemistry, Department of Life and Environmental System Science, Graduate School of NanoBio Sciences, Yokohama City University, Yokohama 236-0027, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ozeki
authors.affiliation: Department of Biochemistry and Molecular Biology, Rajshahi University, Rajshahi 6205, Bangladesh.
authors.forename: A K M
authors.initials: AK
authors.lastname: Asaduzzaman
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry* / Q000494:pharmacology*; D002688:Chitinases / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D052978:Disk Diffusion Antimicrobial Tests; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D010936:Plant Extracts / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D054730:Protein Interaction Domains and Motifs; D012639:Seeds / Q000737:chemistry* / Q000201:enzymology; D013379:Substrate Specificity; D013696:Temperature; D029778:Trichosanthes / Q000737:chemistry*
chemical_list: D000935:Antifungal Agents; D010936:Plant Extracts; D002688:Chitinases
keywords: Amino acid analysis; Antifungal activity; N-terminal sequence