Screen for inhibitors of chitinase

A method of screening test samples for inhibition of chitinase including a primary in vitro enzyme assay and a secondary in vitro enzyme assay, said primary assay comprising the steps of:

• (a) adding test sample to a chitinase producing yeast culture, said culture containing a substrate for chitinase;
• (b) incubating said test sample in said culture under conditions sufficient to detect inhibition of chitinase enzyme activity;
• (c) comparing the extent of substrate conversion in an area around the test sample with the extent of substrate conversion in the area void of said test sample;
• (d.1) observing cell growth of the culture;
• (d.2) determining the presence of chitinase inhibition by observing the extent of substrate conversion;
• and said secondary assay comprising the steps of:
• (e) adding a test sample that exhibits chitinase inhibition in said primary assay to at least two secondary enzyme assays, said secondary enzyme assays selected from the group consisting of a chitinase assay and at least one other control enzyme assays in which the assay detects for the activity of an enzyme other than chitinase
• (f) incubating said test sample in said secondary enzyme assays under conditions sufficient to detect inhibition of chitinase and inhibition of said control enzyme assay;
• (g) simultaneously comparing chitinase inhibition and control enzyme inhibition;
• and
• (h) determining whether the extent of chitinase inhibition is greater than the control enzyme inhibition.