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Lett Appl Microbiol. 2001 Mar;32(3):211-4.

Sensitive plate assay for screening and detection of bacterial polyurethanase activity.

Author information

1
Department of Biological Sciences, South-eastern Louisiana University, Hammond, LA 70402, USA.

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.

PMID:
11264755
[Indexed for MEDLINE]
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