Section 7
Chapter 6,930

3 cyano 4 trifluoromethylumbelliferyl beta d galactopyranoside synthesis properties and application for beta galactosidase assay

Markaryan, A.N.; Voznyi, Y.V.

Analytical Letters 25(4): 681-694


ISSN/ISBN: 0003-2719
Accession: 006929037

Download citation:  

A novel substrate 3-cyano-4-trifluoromethylumbelliferyl-.beta.-D-galactopyranoside, was synthesized and used for the continuous photometric and fluorimetric assay of .beta.-galactosidase from Escherichia coli. The reaction product, 3-cyano-4-trifluoromethylumbelliferone is fluorescent at neutral pH values (excitation and emission wavelength maxima: 440 and 500 nm, respectively) and has intense absorption at 440 nm (t=42000 M-1cm-1, pH 7.5). The properties of the dye released upon the enzyme action on the substrate (low pKa=6.0) and its possible avidity to nitrocellulose paper allowed visual detection up to 0.1 fmoles of .beta.-galactosidase from E. coli in 5 min. The same procedure was used for visual identification of bacterial clones expressing .beta.-galactosidase activity after transferring the lysed clones on nitrocellulose.

PDF emailed within 1 workday: $29.90