L. Fulop et T. Ponyi, RAPID SCREENING FOR ENDO-BETA-1,4-GLUCANASE AND ENDO-BETA-1,4-MANNANASE ACTIVITIES AND SPECIFIC MEASUREMENT USING SOLUBLE DYE-LABELED SUBSTRATES, Journal of microbiological methods, 29(1), 1997, pp. 15-21
Soluble dye-labelled substrates were prepared for the specific and rap
id screening and the measurement of endo-beta-1,4-glucanase and endo-b
eta-1,4-mannanase activities. Soluble carboxy-methylcellulose and locu
st bean galactomannan were dyed with Remazol Brilliant Blue R. A speci
fic screening procedure was developed to isolate genes coding for cell
ulolytic and hemicellulolytic enzymes from a Cellulomonas sp. CelB7 li
brary. The assay is advantageous for the simple, rapid and direct dete
ction of cellulase and hemicellulase activities in a large number of c
lones as found in gene libraries without replica plating or blotting t
echniques. The prepared dye-labelled substrates can also be used for t
he quantitative measurement of specific endo activities of cellulolyti
c and hemicellulolytic enzymes because the undigested substrate can be
precipitated and the amount of soluble dye, determined spectrophotome
trically, is proportional to enzyme activity. (C) 1997 Elsevier Scienc
e B.V.