Y-β-D-GlcX.CHX

• White to pale beige crystalline powder, odorless and hygroscopic.
• Slightly soluble in water (~1-5 mg/mL), highly soluble in DMF/DMSO (>50 mg/mL).

Source

• Chemically synthesized by Koenigs-Knorr glycosylation of 3-indolol with acetobromo-α-D-glucopyranoside, followed by deprotection and salt formation.

Molecular Weight and Structure

• Free acid/base: C14H15NO7, 309.27 g/mol; cyclohexylammonium salt: C20H28N2O7, 408.45 g/mol.
• Features β-D-glucopyranosyl-(1→3)-indolyl linkage; InChI: KUYNOZVWCFXSNE-BYNIDDHOSA-N.

Sugar Specificity

• Selective for β-glucosidase (EC 3.2.1.21), hydrolyzing the β-(1→O)-glycosidic bond.
• Low activity with β-galactosidase or α-glycosidases; Km ≈ 0.3-0.6 mM.

Biological Activity

• Cleavage releases 3-indoxyl, which dimerizes/oxidizes to insoluble indigo (blue, λmax 605 nm) in air.
• Optimal pH 6.8-7.2, 25-37°C; used as renal failure biomarker in plasma.

Purity and Microbial Contamination

• ≥98.5% HPLC purity; <0.5% water, <10 ppm heavy metals; molecular biology grade.
• Endotoxin-free (<1 EU/mg), sterile, no microbial growth per USP <71>.

Identity and Quality Control

• Confirmed by 1H-NMR, HRMS (m/z 310.0924 [M+H]+), [α]D -35° (c=1, H2O), TLC Rf 0.6 (EtOAc:MeOH).
• Functional test: blue color in 1 hr with β-glucosidase; CAS/IR verified.

Shelf Life and Storage

• 36-60 months at -20°C under nitrogen in amber glass; stable 6 months at 4°C in solution.
• Avoid freeze-thaw cycles, light, oxidants; repack in desiccator.

Application

• Chromogenic detection of β-glucosidase in E. coli cloning, plant pathology, soil microbes.
• In situ enzyme histochemistry, HTS inhibitor screening, E. coli enumeration.