Gelatin Hydrolysis Test Introduction Gelatin, a protein derived from the animal protein collagen, has been used

as a solidifying agent in food for a long time. Jello, for example, is made with gelatin. Robert Koch used nutrient gelatin as an early type of solid growth medium. One problem is that many bacteria have the ability to hydrolyze (liquefy) gelatin. This gelatin liquefaction ability (or inability) forms the basis for this test. Results + = Within 7 days of incubation, refrigerated gelatin remains liquid - = After 7 days of incubation, refrigerated gelatin is solid. Methods 1. Obtain two nutrient gelatin tubes (from the back shelf). 2. STAB inoculate one nutrient gelatin tube with your unknown stock culture. Leave the other tube uninoculated (it will act as a control). 3. Incubate at ROOM TEMPERATURE (about 20 C). [This is one of the few tests that are run at suboptimal temperature. The reason is gelatin melts at 28 C. You want to be sure that liquefaction of gelatin is due to bacterial metabolism, not because of incubation temperature]. 4. Every few days, gently place the two tubes in the refrigerator for 15 minutes. Try to minimize agitation of the tubes. 5. After 15 minutes, if the gelatin remains LIQUID, the result is POSITIVE and you may end the test. 6. If the gelatin is SOLID, the result is negative. Continue to incubate the cultures (grow the bacteria some more at room temperature). A NEGATIVE result is only concluded if the gelatin remains solid through 7 days of incubation.