CHITINOLYTIC ENZYMES OF TRICHODERMA-HARZIANUM - PURIFICATION OF CHITOBIOSIDASE AND ENDOCHITINASE

被引:262
作者
HARMAN, GE
HAYES, CK
LORITO, M
BROADWAY, RM
DIPIETRO, A
PETERBAUER, C
TRONSMO, A
机构
[1] CORNELL UNIV, DEPT PLANT PATHOL, GENEVA, NY 14456 USA
[2] CORNELL UNIV, DEPT ENTOMOL, GENEVA, NY 14456 USA
[3] AGR UNIV NORWAY, DEPT BIOTECHNOL SCI, N-1432 AS, NORWAY
[4] UNIV NAPLES, IST PATOL VEGETALE, I-80138 NAPLES, ITALY
[5] CNR, ISTITUENDO CTR TECN LOTTA BIOL, I-80055 PORTICI, ITALY
关键词
BIOCONTROL; BIOLOGICAL CONTROL; MYCOPARASITISM;
D O I
10.1094/Phyto-83-313
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Trichoderma harzianum strain P1 produces a variety of chitinolytic enzymes including N-acetyl-beta-D-glucosaminidases, chitin 1,4-beta-chitobiosidases, and an endochitinase. Chitobiosidases and an endochitinase were purified from dialyzed, concentrated culture filtrates using gel filtration, chromatofocusing, and isoelectric focusing. Three protein bands were evident in the purified chitobiosidase preparation, representing different levels of N-glycosylation of the same protein. The pI of all purified proteins was approximately 3.9. The molecular mass of the principal glycosylated chitobiosidase was 40 kDa, the deglycosylated form was 35 kDa, and the endochitinase was 41 kDa. The chitobiosidases did not react with a polyclonal antibody prepared against the endochitinase, nor did the endochitinase react with an antibody prepared against the chitobiosidase. The endochitinase and the chitobiosidase had broad pH optima. The larger glycosylated chitobiosidase had a 4.0-7.0 pH optimum. The endochitinase optimum activity was at approximately pH 4.0 and gradually declined as pH increased.
引用
收藏
页码:313 / 318
页数:6
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