Howard G T, Vicknair J, MacKie R I
Department of Biological Sciences, South-eastern Louisiana University, Hammond, LA 70402, USA.
Lett Appl Microbiol. 2001 Mar;32(3):211-4. doi: 10.1046/j.1472-765x.2001.00887.x.
A plate assay to screen and detect bacterial polyurethanase in agar medium containing a colloidal polyester-polyurethane and rhodamine B is presented.
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.
The potential advantages of this system are in identification and recovery of viable polyurethanolytic bacteria and quantification of polyurethanase activity.
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.
介绍一种在含有胶体聚酯 - 聚氨酯和罗丹明B的琼脂培养基中筛选和检测细菌聚氨酯酶的平板测定法。
底物水解导致在紫外线照射下可见橙色荧光晕圈的形成。纯化的聚氨酯酶蛋白的聚氨酯酶活性对数与晕圈直径呈线性相关，从而能够对0.81至7.29单位范围内的聚氨酯酶活性进行定量。
该系统的潜在优势在于可鉴定和回收有活性的聚氨酯分解细菌以及对聚氨酯酶活性进行定量。
这些优势主要源于底物与罗丹明B反应水解时观察到的强烈荧光，这允许使用低底物浓度并相应减少检测低水平酶活性所需的时间。
