Efficient chemoenzymatic synthesis of 4-nitrophenyl β-d-apiofuranoside and its use in screening of β-d-apiofuranosidases.

Abstract 4-Nitrophenyl β- d -apiofuranoside as a chromogenic probe for detection of β- d -apiofuranosidase activity was prepared in 61% yield from 2,3-isopropylidene-α,β- d -apiofuranose through a sequence of five reactions. The synthesis involves one regioselective enzymatic step—benzoylation of primary hydroxyl of 2,3-isopropylidene-α,β- d -apiofuranose catalysed by Lipolase 100T and stereoselective β- d -apiofuranosylation of p -nitrophenol using BF 3 ⋅OEt 2 /Et 3 N. The product was used for screening of β- d -apiofuranosidase activity in 61 samples of crude commercial enzymes and plant materials. Fifteen enzyme preparations originating from different strains of genera Aspergillus display β- d -apiofuranosidase activity. The highest activity was found in Rapidase AR 2000 (78.27 U/g) and lyophilized Viscozyme L (64,36 U/g).