An improved method for detection and quantification of chitinase activities.
Chitinases are enzymes that serve critical roles in fungal growth and development, in resistance of plants to fungal pathogens, and in parasitism of insects by entomopathogenic fungi. The term "chitinase" is used for 3 enzymatic activities: N-acetylglucosaminidases, which sequentially release N-acetylglucosamine residues from the chitin polymer; chitobiosidases, which release disaccharides; and endochitinases, which cleave within the polymer and release oligosaccharides. We describe a technique where chitinases are separated on non-denaturing polyacrylamide gels, activities are visualized and characterized with chitinase specific substrates, and specific activities are estimated by image analysis. This technique permits a rapid determination of all of the types of chitinases present within a sample as well as their activities.
Guthrie
Jennifer L
JL
Department of Biological Sciences, Brock University, St. Catharines, ON L2S 3A1, Canada.
Khalif
Sagal
S
Castle
Alan J
AJ
D000364:Agaricus / Q000201:enzymology; D002688:Chitinases / Q000032:analysis / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006923:Hymecromone / Q000031:analogs & derivatives; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology
D006923:Hymecromone; D002688:Chitinases