Literature DB >> 17434593

A modified method for the detection of microbial proteases on agar plates using tannic acid.

Saurabh Saran1, Jasmine Isar, Rajendra Kumar Saxena.   

Abstract

In routine assay for the screening of microbes producing proteases, 10% trichloroaceticacid (TCA) is flooded on the milk agar plates after inoculation and required incubation to precipitate the protein. However, the clarity of the hydrolyzed zone is not very sharp and distinct. We herein present an improved assay for detecting the presence of extracellular protease from microorganisms on agar plates. In this method 10% tannic acid is flooded on the milk agar plate (in place of, TCA) to observe the zone of hydrolysis. Tannic acid sharply increases the colour intensity of the plate, as it favours the precipitation of the unhydrolyzed protein in the plate, thereby improving the contrast between the intact zones and the enzymatic lyses zones of the substrate. Our results indicate that this method is useful to detect extracellular proteases produced by both fungi as well as bacteria. The method used in the present study is sensitive, and can be easily performed for screening of large number of microbial cultures. This is the first report on the use of tannic acid for the detection of microbial proteases.

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Year:  2007        PMID: 17434593     DOI: 10.1016/j.jbbm.2007.03.005

Source DB:  PubMed          Journal:  J Biochem Biophys Methods        ISSN: 0165-022X


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