Professor
Characterization of a Yam Class IV Chitinase Produced by the Recombinant Pichia pastoris X-33
Department of Botany, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh
Abbreviations: Endo H, Endoglycosidase H; PR protein, pathogenesis related protein; P. pastoris, Pichia pastoris; VTS, vacuolar targeting signal; PCR, polymerase chain reaction; PAGE, polyacrylamide gel electrophoresis; SDS, sodium dodecyl sulfate
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 yields remarkably, such as 66 mg/ L of culture medium. The chitinase was purified by column chromatography after Endoglycosidase H treatment and was then characterized. The Pichia-produced chitinase It showed properties that are similar to the original chitinase E purified from the yam tuber, as reported by Arakane et al. (2000). This Pichia-produced chitinase showed also strong lytic activity against Fusarium oxysporum and Phytophthora nicotianae, wide pH and thermal stability, optimum activity at higher temperature such as 70oC, and high substrate affinity, thus indicating that we one can use this Pichia-produced yam chitinase as a bio-control agent.
Key words: yam (Dioscorea oppsita Thunb); class IV chitinase; bio-control agent; recombinant Pichia pastoris; lytic activiy
Introduction
Chitinases [EC 3.2.1.14] are diversified in biological functions, depending on sources. Chitinases, which hydrolyze chitin, occur in a wide range of organisms, including viruses, bacteria, fungi, insects, higher plants, and animals.1) Bacterial chitinases play roles in nutrition and parasitism while fungal chitinases have autolytic, nutritional, and morphogenetic roles. Viral chitinases are involved in pathogenesis.2) Invertebrate chitinases play roles in digestion. In insects and crustaceans, chitinases are associated with ecdysis through degradation of old cuticle.3) Some
Department of Botany, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh
Abbreviations: Endo H, Endoglycosidase H; PR protein, pathogenesis related protein; P. pastoris, Pichia pastoris; VTS, vacuolar targeting signal; PCR, polymerase chain reaction; PAGE, polyacrylamide gel electrophoresis; SDS, sodium dodecyl sulfate
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 yields remarkably, such as 66 mg/ L of culture medium. The chitinase was purified by column chromatography after Endoglycosidase H treatment and was then characterized. The Pichia-produced chitinase It showed properties that are similar to the original chitinase E purified from the yam tuber, as reported by Arakane et al. (2000). This Pichia-produced chitinase showed also strong lytic activity against Fusarium oxysporum and Phytophthora nicotianae, wide pH and thermal stability, optimum activity at higher temperature such as 70oC, and high substrate affinity, thus indicating that we one can use this Pichia-produced yam chitinase as a bio-control agent.
Key words: yam (Dioscorea oppsita Thunb); class IV chitinase; bio-control agent; recombinant Pichia pastoris; lytic activiy
Introduction
Chitinases [EC 3.2.1.14] are diversified in biological functions, depending on sources. Chitinases, which hydrolyze chitin, occur in a wide range of organisms, including viruses, bacteria, fungi, insects, higher plants, and animals.1) Bacterial chitinases play roles in nutrition and parasitism while fungal chitinases have autolytic, nutritional, and morphogenetic roles. Viral chitinases are involved in pathogenesis.2) Invertebrate chitinases play roles in digestion. In insects and crustaceans, chitinases are associated with ecdysis through degradation of old cuticle.3) Some
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