Semiquantitative Plate Assay for Determination of Cellulase Production by Trichoderma viride.
A plate clearing assay was devised to screen for high-producing cellulase mutants of Trichoderma viride. The method employs (i) the use of either rose bengal or oxgall to limit colony size and (ii) Phosfon D (tributyl-2, 4-dichloroben-zylphosphonium chloride) to enhance cellulase detection, in combination with acid-swollen cellulose on agar plates. The method was used to isolate constitutive cellulase mutants of T. viride and should prove useful for isolating high-producing mutants from a range of organisms. This technique has been also used to determine the concentration at which glucose and glycerol inhibit cellulase synthesis by catabolite repression in the wild-type strains.
Montenecourt
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BS
Department of Biochemistry and Microbiology, Cook College, Rutgers - The State University of New Jersey, New Brunswick, New Jersey 08903.
Eveleigh
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DE