Purpose: To determine whether a bacterium produces a lipase that will hydrolyze a neutral fat to fatty acid and glycerol.Principle: The lipid tributyrin is emulsified into water using Tween 80 and ultrasonification. The acid base indicator spirit blue is also present in the medium. If tributyrin is hydrolyzed via a lipase, the result will be fatty acid and glycerol. The localization of fatty acid will cause the colony to take on a bluish hue.The plate to the right illusrates possible reactions observed on Sprit Blue medium.A. Hydrolysis of lipid (notice blue tint to colony) B. Absense of lipid hydrolyis |
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