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  • Rapid detection of total and fecal coliforms in water by enzymatic hydrolysis of 4-methylumbelliferone-beta-D-galactoside.

Rapid detection of total and fecal coliforms in water by enzymatic hydrolysis of 4-methylumbelliferone-beta-D-galactoside.

Applied and environmental microbiology (1988-08-01)
J D Berg, L Fiksdal
ABSTRACT

Three fluorogenic methylumbelliferone (MU) substrates were evaluated for rapid detection of total and fecal coliform bacteria (TC and FC) in drinking water. 4-MU-beta-D-galactoside, MU-heptanoate, and MU-glucuronide were used to determine enzyme activity as a surrogate measure of coliform concentration. Coliforms occurring in river water and in potable water artificially contaminated with raw sewage were tested. The initial rate of hydrolysis (delta F) of MU-beta-D-galactoside showed promise as an indicator of TC and FC within 15 min. delta F of MU-glucuronide was insufficient in the 15-min assay, and combinations of the MU substrates did not enhance delta F. A direct membrane filter method incorporating MU-beta-D-galactoside into an agar medium allowed the detection of as few as 1 FC per 100 ml within 6 h.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
4-Methylumbelliferyl α-D-galactopyranoside, ≥98% (TLC)
Millipore
m-Endo Broth, NutriSelect® Plus, suitable for microbiology
Sigma-Aldrich
4-Methylumbelliferyl β-D-galactopyranoside, ≥99% (TLC)
Sigma-Aldrich
4-Methylumbelliferyl heptanoate, ≥95% (GC)