title: Characterization of a yam class IV chitinase produced by recombinant Pichia pastoris X-33.
abstract: A yam (Dioscorea opposita Thunb) class IV chitinase, whose genomic DNA was cloned by Mitsunaga et al. (2004), was produced by the recombinant Pichia pastoris X-33 in high yields such as 66 mg/L of culture medium. The chitinase was purified by column chromatography after Endoglycosidase H treatment and then characterized. It showed properties similar to the original chitinase E purified from the yam tuber reported by Arakane et al. (2000). This Pichia-produced chitinase also showed strong lytic activity against Fusarium oxysporum and Phytophthora nicotianae, wide pH and thermal stability, optimum activity at higher temperature such as 70 °C, and high substrate affinity, indicating that one can use this Pichia-produced yam chitinase as a bio-control agent.
authors.affiliation: a The United Graduate School of Agricultural Sciences , Tottori University , Tottori , Japan.
authors.forename: Muhammad Ali
authors.initials: MA
authors.lastname: Akond
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Matsuda
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ishimaru
authors.forename: Ken
authors.initials: K
authors.lastname: Iwai
authors.forename: Akira
authors.initials: A
authors.lastname: Saito
authors.forename: Akio
authors.initials: A
authors.lastname: Kato
authors.forename: Shuhei
authors.initials: S
authors.lastname: Tanaka
authors.forename: Jun
authors.initials: J
authors.lastname: Kobayashi
authors.forename: Daizo
authors.initials: D
authors.lastname: Koga
mesh_terms: D000595:Amino Acid Sequence; D061046:Biological Control Agents; D002688:Chitinases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D046508:Culture Techniques; D004274:DNA, Recombinant / Q000235:genetics*; D027742:Dioscorea / Q000201:enzymology* / Q000235:genetics*; D004795:Enzyme Stability; D005670:Fusarium / Q000187:drug effects; D005818:Genetic Engineering; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010838:Phytophthora / Q000187:drug effects; D010843:Pichia / Q000235:genetics* / Q000254:growth & development; D017421:Sequence Analysis; D013696:Temperature
chemical_list: D061046:Biological Control Agents; D004274:DNA, Recombinant; D002688:Chitinases
keywords: bio-control agent; class IV chitinase; lytic activiy; recombinant Pichia pastoris; yam (Dioscorea oppsita Thunb)