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Fluorogenic and chromogenic glycosides as substrates and ligands of carbohydrases

1988; Academic Press; Linguagem: Inglês

10.1016/0076-6879(88)60106-6

1557-7988

Herman van Tilbeurgh, Frank G. Loontiens, Clement K. De Bruyne, Marc Claeyssens,

Enzyme Catalysis and Immobilization

Nitrophenylglycosides are frequently used substrates of carbohydrases. Alternatively, the 4-methylumbelliferyl (7-hydroxy-4-methylcoumaryl) derivatives offer a more sensitive (fluorometric) method of detection. Some of these compounds have become commercially available. This chapter describes the preparation and use of these glycosides in the study of some cellulolytic enzymes. The difference in spectral properties of free 4-methylumbelliferone and its carbohydrate conjugates allows sensitive and continuous assays of cellulolytic activities in absorption or fluorescence modes. The preparation and use of 1-thio derivatives with different chromophoric reporter groups are included. Due to their optical characteristics, the chromophoric (fluorochromic) derivatives offer distinct advantages over the use of classical substrates of cellulolytic enzymes, as they are sensitive tools for the determination of the number of binding sites, study of association modes, and binding kinetics, breakdown patterns, and inhibition characteristics. The chapter describes some applications of chromophoric.