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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

1992

ISSN/ISBN: 0003-2719
Accession: 006929037
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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.

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Related References

Yegorov, A.M.; Markaryan, A.N.; Vozniy, Y.V.; Cherednikova, T.V.; Demcheva, M.V.; Berezin, I.V. 1988: 4 trifluoromethylumbelliferyl beta d galactopyranoside its synthesis and application as the fluorigenic substrate of beta galactosidase escherichia coli for screening monoclonal antibodies by immunosorbent elisa Analytical Letters 21(2): 193-210
Markaryan, A.N.; Voznyi, Y.V. 1992: 3-Cyano-4-trifluoromethylumbelliferyl--D-galactopyra-noside: Synthesis, Properties and Application for -Galactosidase Assay Analytical Letters 25(4): 681-694
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