Sensitive plate assay for screening and detection of bacterial polyurethanase activity

G. T. Howard, J. Vicknair, R. I. Mackie

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)211-214
Number of pages4
JournalLetters in Applied Microbiology
Volume32
Issue number3
DOIs
StatePublished - 2001

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

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