中国生物工程杂志

In order to improve the accuracy and sensitivity in detecting of chitinase isozymes from Trichoderma. spp, a rapid sensitive method is established. It was employed to detect chitinase isozymes from Trichoderma viride LTR-2 through native gel electrophoresis, denatured gel electrophoresis and in situ gel electrophoresis with Calcofluor white M2R. Two chitinase isozymes native bands CHI65 and CHI42 were revealed in solid plate containing chitin and Calcofluor white M2R with native gel electrophoresis after chitin grown culture was concentrated 5 times. The two bands showed a smear instead of well-defined band with in situ gel electrophoresis after 20 times concentration. While only CHI65 chitinase was found with denatured gel electrophoresis after 10 times concentration. The molecular weight of CHI65 and CHI42 was separately about 65KDa and 42KDa. As a result, it was suitable for detecting chitinase isozymes through active gel electrophoresis with Calcofluor white M2R. The result showed that it was effective for differentiation among chitinase isozymes when total chitinase loading amount was 0.47U.