Use of Coomassie blue-polyurethane interaction in screening of polyurethanase proteins and polyurethanolytic bacteria

The interaction of the dye Coomassie blue with intact polyurethane provides the basis for a rapid and sensitive assay system for bacterial strains pos ing polyurethanase activity. The potential advantage of this system in enum erating and characterizing polyurethanolytic bacteria derives largely from the intense color of the dye-substrate complex, which allows the use of ver y low substrate concentrations and a corresponding decrease in the time req uired to detect lower levels of enzyme activity. The method entails incubat ion of agar plates containing the polyurethane substrate, followed by immer sion in 0.1% Coomassie blue then destaining with 10% acetic acid-40% methan ol. Resulting agar plates contain transparent bands corresponding to enzyma tic activity against a blue background. (C) 1999 Elsevier Science Ltd. All rights reserved.