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Sensitive plate assay for screening and detection of bacterial polyurethanase activity
Author(s) -
Howard G.T.,
Vicknair J.,
Mackie R.I.
Publication year - 2001
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2001.00887.x
Subject(s) - library science , biological sciences , history , biology , microbiology and biotechnology , computer science
G.T. HOWARD, J. VICKNAIR AND R.I. MACKIE. 2001.Aims: A plate assay to screen and detect bacterial polyurethanase in agar medium containing a colloidal polyester‐polyurethane and rhodamine B is presented. Methods and Results: Substrate hydrolysis causes the formation of orange fluorescent halos visible upon u.v. irradiation. The logarithm of polyurethanase activity from a purified polyurethanase protein is linearly correlated with the diameter of halos, thereby allowing quantification of polyurethanase activities ranging from 0·81 to 7·29 Units. Conclusions: The potential advantages of this system are in identification and recovery of viable polyurethanolytic bacteria and quantification of polyurethanase activity. Significance and Impact of the Study: These advantages are derived largely from the intense fluorescence observed due to the hydrolysis of substrate reacting with rhodamine B allowing for the use of low substrate concentrations and corresponding decrease in time required detecting low levels of enzyme activity.