Immobilized Recombinant Chitinase and its Inhibition Effect on Botrytis Cinerea

Abstract

To isolate recombinant chitinase quickly and boost its anti-fungi activities in vitro, functional magnetic nanometer carrier was used to immobilize recombinant chitinase from the crude enzyme solution and immobilized recombinant chitinase was applied to test whether it would inhibit the growth of gray mold from fruits. In this study, the carboxyl magnetic carrier was produced by solvent thermal reduction method and characterized by scanning electron microscope (SEM) and fourier transform infrared spectrometer (FTIR). Then, the carboxyl magnetic carrier activated by EDC/NHS was applied to immobilize recombinant chitinase and the immobilization efficiency was investigated by quantitative analysis. To obtain the highest immobilization efficiency, reaction conditions were optimized through combining different pH, temperature and reaction period. The results show that the surface of magnetic carrier was successfully carboxyl and the average diameter was 200nm. The immobilization efdiciency could reach the peak 64.43% after 7h reaction at the condition of pH 6 and 25°C. It also shows that immobilized recombinant chitinase can significantly inhibit the growth of gray mold isolated from table grape compared with the enzyme without immobilization with magnetic nanometer carrier.