Sensitive plate assay for screening and detection of bacterial polyurethanase activity.
OBJECTIVE
A plate assay to screen and detect bacterial polyurethanase in agar medium containing a colloidal polyester-polyurethane and rhodamine B is presented.


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.


CONCLUSIONS
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.
Howard
G T
GT
Department of Biological Sciences, South-eastern Louisiana University, Hammond, LA 70402, USA.
Vicknair
J
J
MacKie
R I
RI
D001426:Bacterial Proteins; D005453:Fluorescence; D008049:Lipase / Q000032:analysis*; D011140:Polyurethanes / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology*; D012235:Rhodamines / Q000378:metabolism
D001426:Bacterial Proteins; D011140:Polyurethanes; D012235:Rhodamines; C408182:pueA protein, Pseudomonas chlororaphis; D008049:Lipase; C029773:rhodamine B