Table 10.1 Glycosidase enzymes and their fluorogenic and chromogenic substrates.

Carbohydrate
(Enzyme)
Notes on Enzyme Activity Labeled Substrate (Abs/Em of the products) * Cat #
β-D-Galactopyranoside
(β-Galactosidase, E.C. 3.2.1.23)
  • Useful as a reporter gene marker ref
  • Useful for ELISAs ref
  • Useful for enumerating coliforms from the family Enterobacteriaceae ref
  • Useful for classifying mycobacteria ref
Blue-fluorescent product 4-Methylumbelliferyl (360/449) ref M1489MP
Green-fluorescent product Fluorescein (490/514) ref F1179
5-(Pentafluorobenzoylamino)-fluorescein (490/514) P11948
C2-Fluorescein (490/514) ref A22010
C12-Fluorescein (490/514) ref D2893, I2904 ‡
5-Chloromethylfluorescein (490/514) ref D2920 †
Red-fluorescent product C12-Resorufin (571/585) ref I2906 ‡
DDAO (646/659) D6488
Resorufin (571/585) ref R1159
Chromogenic substrate 5-Bromo-4-chloro-3-indoyl (615/NA) ref X-Gal; B1690
β-D-Glucopyranoside (β-Glucosidase, E.C. 3.2.1.21)
  • Deficiency in acid β-glucosidase, which leads to abnormal lysosomal storage, characterizes Gaucher disease ref
  • Useful as a marker for the endoplasmic reticulum ref
Green-fluorescent product Fluorescein (490/514) ref F2881
5-(Pentafluorobenzoylamino)fluorescein (490/514) ref P11947
β-D-Glucuronide (β-Glucuronidase, GUS; E.C. 3.2.1.31)
  • Useful as a reporter gene marker ref
  • Useful as a lysosomal marker ref
  • Useful for detecting E. coli; 94–96% of E. coli contain this enzyme, but it is less common in Shigella (44–58%), Salmonella (20–29%) and Yersinia strains ref
Blue-fluorescent product 4-Methylumbelliferyl (360/449) ref M1490
Green-fluorescent product Fluorescein (490/514) ref F2915
5-(Pentafluorobenzoylamino)fluorescein (490/514) ref P11949
C12-Fluorescein (490/514) ref I2908 ‡
Chromogenic substrate 5-Bromo-4-chloro-3-indoyl (615/NA) ref X-GlcU, B1691
* Approximate absorption (Abs) and fluorescence emission (Em) maxima of enzymatic hydrolysis product, in nm. † DetectaGene Green Gene Expression Kit. ‡ ImaGene Green or ImaGene Red Gene Expression Kit. NA = Not applicable.