p-Nitrophenyl beta-D-glucuronide → 4-Nitrophenyl--d-glucopyranoside

HRID 1807365Notes

Reaction equation

HRID 1807365 reaction equation
[CH:1]1[C:6]([N+:7]([O-:9])=[O:8])=[CH:5][CH:4]=[C:3]([O:10][C@@H:11]2[O:16][C@H:15]([C:17](O)=[O:18])[C@@H:14]([OH:20])[C@H:13]([OH:21])[C@H:12]2[OH:22])[CH:2]=1>P([O-])([O-])([O-])=O.[Na+].[Na+].[Na+]>[O:10]([C:3]1[CH:2]=[CH:1][C:6]([N+:7]([O-:9])=[O:8])=[CH:5][CH:4]=1)[CH:11]1[O:16][C@H:15]([CH2:17][OH:18])[C@@H:14]([OH:20])[C@H:13]([OH:21])[C@H:12]1[OH:22]

Participants

3

Reactant 1

HCID 82565p-Nitrophenyl beta-D-glucuronide2×

Product 1

HCID 111973694-Nitrophenyl--d-glucopyranoside

Reagents, catalysts & solvents 1

HCID 24243Trisodium PhosphateSolvent

Procedure

A colorimetric pNPG (p-nitrophenyl-(β-D-glucopyranoside)-based assay was used for measuring β-glucosidase activity. In a total volume of 100 μL, 20 μL clear media supernatant containing β-glucosidase enzyme was added to 4 mM pNPG (Sigma-Aldrich, Inc. St. Louis, Mo.) solution in 50 mM sodium phosphate buffer at pH6.5. The reactions were incubated at pH 6.5, 45° C. for 1 hour. The reaction mixture was quenched with 100 μL of 1M sodium carbonate pH 11 solution. The absorbance of the solution was measured at 405 nm to determine the conversion of pNPG to p-nitrophenol. The release of p-nitrophenol (ε=17,700 M−1 cm−1) was measured at 405 nm to calculate β-glucosidase activity. Detectable β-glucosidase activity was observed under high throughput screening conditions (pH 7, 50° C.).

Workup

  1. customThe reaction mixture was quenched with 100 μL of 1M sodium carbonate pH 11 solution
  2. customhigh throughput screening conditions (pH 7, 50° C.)

Sources & evidence

ORD record
ord-dc44068fe7964147a512d36e1023d785
Source dataset
uspto-grants-2013_01
Patent number
US08357523B2

Reaction notes

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