3-Cyano-4-trifluoromethylumbelliferyl-β-D-galactopyra-noside: Synthesis, Properties and Application for β-Galactosidase Assay
| Autor: | Yakov V. Voznyi, Adam N. Markaryan |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
chemistry.chemical_classification
Lysis Chromatography Bicyclic molecule Stereochemistry Biochemistry (medical) Clinical Biochemistry Glycoside Substrate (chemistry) Biochemistry Fluorescence Analytical Chemistry chemistry.chemical_compound chemistry Electrochemistry Absorption (chemistry) Nitrocellulose Spectroscopy Lactone |
| Zdroj: | Analytical Letters. 25:681-694 |
| ISSN: | 1532-236X 0003-2719 |
| DOI: | 10.1080/00032719208020027 |
| Popis: | A novel substrate 3-cyano-4-trifluoromethylumbel-liferyl-β-D-galactopyranoside, was synthesized and used for the continuous photometric and fluorimetric assay of β-galactosidase from E.coli. The reaction product, 3-cyano-4-triluoromethylumbelliferone is fluorescent at neutral pH values (excitation and emission wavelength maxima: 440 and 500 nm, respectively and has intense absorption at 440 nm (c=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 β-galactosidase from E.coli in 5 min. The same procedure was used for visual identification of bacterial clones expressing β-galactosidase activity after transferring the lysed clones on nitrocellulose. |
| Databáze: | OpenAIRE |
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