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indole production testTryptophan is an essential amino acid that can undergo oxidation by way of the enzymatic activities of some bacteria.Conversion of tryptophan into metabolic products is mediated by the enzyme tryptophanase.This ability to hydrolyze trytophan with the production of indole Is not a characteristic of all microorganisms and therefore serves as a biochemical marker. In this experiment,SIM agar,which contains the substrate tryptophan,is used.The presence of indole is detectable by adding kovac's reagent,which produces a cherry red reagent layer.This color is produced by the reagent,which is composed of dimethylaminobezaldehyde,butanol,and hydrochloric acid.Indole is extracted from the medium into the reagent layer by the acidified butyl alcohol component and forms a complex with the dimethylaminobenzaldehyde,yielding the cherry red color. Cultures producing a red reagent layer following addition of kovac's reagent are indole positive.The absence of red coloration demonstrates that the substrate tryptophan was not hydrolyzed and indicates an indole negative reaction.MediaSIM agar deep tubes per designated student group;4 for the short version,14 for the long version.EquipmentBunsen burner,inoculating needle,test tube rack,and glassware marking pencil.
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