Resorufin-Glc (Resorufin β-D-glucopyranoside)

Resorufin β-D-glucopyranoside (CAS 101490-85-1) is a synthetic, nonfluorescent fluorogenic substrate specifically designed for ultrasensitive detection of β-glucosidase enzyme activity through enzymatic hydrolysis that liberates the intensely red-fluorescent resorufin dye (Ex/Em 571/585 nm, ε=56,000 M⁻¹cm⁻¹, φ=0.74). With molecular formula C18H17NO8 (MW 375.33 g/mol), this chromogenic probe features the resorufin phenoxazinone aglycone conjugated via a β-1-O-glycosidic linkage to D-glucopyranose, remaining stable at physiological pH (6.5-8.0) until cleaved by β-glucosidases (EC 3.2.1.21) such as those from Clostridium cellulovorans, Trichoderma reesei, or human lysosomal acid-β-glucosidase (GBA/glucocerebrosidase), yielding >1000-fold fluorescence enhancement for real-time kinetic assays. Optimal for high-throughput screening (HTS) of glucosidase inhibitors, diagnostics of Gaucher disease (GBA deficiency), and functional analysis of microbial β-glucosidases in biofuel/cellulose degradation research, it outperforms p-nitrophenyl-β-glucoside (pNP-Glc) by providing fluorescence-based ratiometric readout with nanomolar enzyme sensitivity and no interference from yellow chromophores. Cell-permeable in DMSO stocks, it enables live-cell monitoring of lysosomal GBA activity, plant cell wall hydrolase kinetics, and directed evolution libraries for engineered glucosidases, while its orthogonality to GFP/FRET pairs supports multiplexing in 384-well formats for drug discovery targeting carbohydrate metabolism disorders, cancer glycoenzymes, and microbiome functional screening.

Appearance

• Off-white to pale yellow crystalline powder or solid.
• Non-fluorescent until β-glucosidase cleavage.

Source

• Koenigs-Knorr synthesis: protected β-D-glucopyranosyl bromide + resorufin.
• Commercial suppliers: Sigma-Aldrich (≥90%), Abcam (≥95%).

Molecular Weight and Structure

• Formula: C18H17NO8; MW 375.33 g/mol.
• 7-O-(β-D-glucopyranosyl)resorufin (β1-glycosidic linkage).​

Sugar Specificity

• Highly selective β-D-glucopyranoside for β-glucosidases (EC 3.2.1.21).
• 500x preference over β-galactosidases/α-glucosidases.​

Biological Activity

• Fluorogenic substrate; no intrinsic bioactivity.
• Detects GBA activity in Gaucher disease diagnostics (pM sensitivity).​

Purity and Microbial Contamination

• ≥90-98% (HPLC); analytical grade >95%.
• Synthetic reagent; non-sterile.​

Identity and Quality Control

• ¹H-NMR (anomeric H-1′ 5.15 ppm d, J=7.5 Hz); HRMS m/z 376 [M+H]+.
• Post-hydrolysis Ex/Em 571/585 nm confirmation.​

Shelf Life and Storage

• 2-3 years at −20°C desiccated; DMSO stock 20 mg/mL stable 6 months −80°C.
• Light/moisture sensitive; avoid freeze-thaw.​

Applications

• β-Glucosidase HTS inhibitor screening; Gaucher disease diagnostics.
• Microbial cellulase assays; live-cell lysosomal GBA monitoring.

Key Characteristics

• 1000x fluorescence enhancement; no-wash kinetic assays.
• Red emission orthogonal to GFP; nanomolar enzyme detection.​

Citation Links

• https://www.abcam.com/en-us/products/biochemicals/resorufin-b-d-glucopyranoside-ab274311​
• https://www.sigmaaldrich.com/US/en/product/sigma/r4758​
• https://www.chemdad.com/index.php?c=article&id=2124​
• https://pubchem.ncbi.nlm.nih.gov/compound/13558997​
• https://www.chemicalbook.com/ChemicalProductProperty_EN_CB6278108.htm​
• https://www.chemsrc.com/en/cas/101490-85-1_345667.html​
• https://amp.chemicalbook.com/ProductChemicalPropertiesCB6278108_EN.htm​
• https://pubchem.ncbi.nlm.nih.gov/compound/Resorufin-galactopyranoside​
• https://www.abcam.com/en-us/products/biochemicals/resorufin-d-galactopyranoside-ab273639​
• https://pubchem.ncbi.nlm.nih.gov/compound/Resorufin