Hydrolysis of plant cuticle by plant pathogens. Properties of cutinase I, cutinase II, and a nonspecific esterase isolated from Fusarium solani pisi.
The properties of the homogeneous cutinase I, cutinase II, and the nonspecific esterase isolated from the extracellular fluid of cutin-grown Fusarium solani F. pisi (R.E. Purdy and P.E. Kolattukudy (1975), Biochemistry, preceding paper in this issue) were investigated. Using tritiated apple cutin as substrate, the two cutinases showed similar substrate concentration dependence, protein concentration dependence, time course profiles, and pH dependence profiles with optimum near 10.0. Using unlabeled cutin, the rate of dihydroxyhexadecanoic acid release from apple fruit cutin by cutinase I was determined to be 4.4 mumol per min per mg. The cutinases hydrolyzed methyl hexadecanoate, cyclohexyl hexadecanoate, and to a much lesser extent hexadecyl hexadecanoate but not 9-hexadecanoyloxyheptadecane, cholesteryl hexadecanoate, or hexadecyl cinnamate. The extent of hydrolysis of these model substrates by cutinase I was at least three times that by cutinase II. The nonspecific esterase hydrolyzed all of the above esters except hexadecyl cinnamate, and did so to a much greater extent than did the cutinases. None of the enzymes hydrolyzed alpha- or beta-glucosides of p-nitrophenol. p-Nitrophenyl esters of fatty acids from C2 through C18 were used as substrates and V's and Kms were determined...
D001665:Binding Sites / Q000187:drug effects; D002265:Carboxylic Ester Hydrolases; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D005670:Fusarium / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000032:analysis*; D006868:Hydrolysis; D006880:Hydroxy Acids / Q000378:metabolism; D007527:Isoenzymes / Q000032:analysis*; D007531:Isoflurophate / Q000494:pharmacology; D007700:Kinetics; D050356:Lipid Metabolism; D010261:Paraoxon / Q000494:pharmacology; D010944:Plants / Q000382:microbiology*; D011108:Polymers / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D013329:Structure-Activity Relationship; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D006880:Hydroxy Acids; D007527:Isoenzymes; D011108:Polymers; D013439:Sulfhydryl Reagents; D007531:Isoflurophate; D012694:Serine; D006867:Hydrolases; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D010261:Paraoxon