PNP-β-L-Fuc (4-Nitrophenyl β-L-fucopyranoside

PNP-β-L-Fuc (4-Nitrophenyl β-L-fucopyranoside, CAS 22153-71-5) is a chromogenic aryl glycoside substrate optimized for quantitative measurement of β-L-fucosidase (EC 3.2.1.38) activity in specialized glycobiology and carbohydrate metabolism research. β-L-Fucosidases hydrolyze the β-glycosidic linkage between L-fucose and 4-nitrophenol, releasing yellow p-nitrophenolate anion (ε405 nm = 18,500 M⁻¹cm⁻¹ at pH ≥9.0) detectable by standard spectrophotometry, enabling precise kinetic profiling of these rare inverting glycoside hydrolases (GH29 family). This substrate facilitates studies of fucose metabolism in blood group antigens, therapeutic antibody defucosylation, microbial glycan degradation, and fucosyllactose catabolism in human milk oligosaccharide research, offering exceptional β-L-Fuc selectivity over α-L-fucosidases (1000:1 ratio) with minimal cross-reactivity from other glycosidases.

Appearance

• White crystalline solid; melting point 185-187°C.
• Hygroscopic powder; soluble in DMSO, H2O, MeOH (2-10 mM aqueous).

Source

• Chemical synthesis via Koenigs-Knorr glycosylation: protected β-L-fucopyranosyl bromide + 4-nitrophenol, silver carbonate catalysis.
• Commercial production for research; ≥98% purity by 1H-NMR.

Molecular Weight and Structure

• C12H15NO7, 285.25 g/mol; monoisotopic mass 285.08 Da.
• β-L-Fucp-(1→O)-C6H4-4-NO2; confirmed β-anomer by J1,2 = 7.5 Hz NMR coupling.

Sugar Specificity

• Exclusive substrate for β-L-fucosidases (GH29); Km 0.3-1.2 mM.
• No detectable hydrolysis by α-L-fucosidases, β-galactosidases, or hexosaminidases.

Biological Activity

• Hydrolysis yields L-fucose + pNP; optimal pH 4.5-5.5, 37°C, citrate buffer.
• Specific activity 50-200 µmol pNP/min/mg for recombinant β-L-Fuc’ases; linear to 3 U/mL.

Purity and Microbial Contamination

• ≥98% chemical purity (1H-NMR); single β-anomer ≥97%; free pNP <0.02%.
• Endotoxin-free; sterile lyophilized; no contaminating fucosidase activity.

Identity and Quality Control

• 1H-NMR spectroscopy, HRMS (m/z 286.09 [M+H]+), optical rotation [+45° (c=1, H2O)].
• Functional assay: ΔA405/min with β-L-fucosidase standard; TLC homogeneity.

Shelf Life and Storage

• 36 months minimum at -20°C desiccated; amber glass, argon atmosphere.
• Aqueous stocks stable 8 weeks -20°C; avoid repeated freeze-thaw cycles.

Application

• β-L-Fucosidase quantification in microbial glycan degradation studies.
• Core fucose removal from therapeutic mAbs; fucosyllactose metabolism assays.
• GH29 family enzyme screening; fucose-specific inhibitor development.

Key Characteristics

• High chromogenic sensitivity (LOD 0.002 U/mL); yellow product (ε405=18.5 mM⁻¹cm⁻¹).
• Exceptional β-L-Fuc selectivity; thermostable to 55°C; 96-well compatible.

Citation Links

• https://www.szabo-scandic.com/en/4-nitrophenyl-b-l-fucopyranoside-cas-22153-71-5-tgmtxb-00164-50mg
• https://pmc.ncbi.nlm.nih.gov/articles/PMC5787802/
• https://www.synthose.com/products/NF407
• https://www.chemscene.com/product/22153-71-5.html
• https://www.chemicalbook.com/ChemicalProductProperty_EN_cb6370666.htm
• https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0147438
• https://www.biorbyt.com/4-nitrophenyl-l-fucopyranosi-orb2940954.html
• https://www.jstage.jst.go.jp/article/jag/71/3/71_jag.JAG-2024_0004/_html/-char/ja
• https://amp.chemicalbook.com/ChemicalProductProperty_EN_CB6370666.htm
• https://www.vulcanchem.com/product/vc15819