CAS 369-07-3|ONPG

Introduction:Basic information about CAS 369-07-3|ONPG, including its chemical name, molecular formula, synonyms, physicochemical properties, and safety information, etc.
Common NameONPG
CAS Number369-07-3Molecular Weight301.249
Density1.6±0.1 g/cm3Boiling Point572.4±50.0 °C at 760 mmHg
Molecular FormulaC12H15NO8Melting Point186-193 °C (dec.)
MSDSChineseUSAFlash Point300.0±30.1 °C

Names

Name2-Nitrophenyl-beta-D-galactopyranoside
SynonymMore Synonyms

ONPG BiologicalActivity

DescriptionONPG is a colorimetric and spectrophotometric substrate for detection of β-galactosidase activity.
Related CatalogSignaling Pathways >>Others >>OthersBiochemical Assay ReagentsResearch Areas >>Others
In VitroThe enzyme displays high hydrolysis ability for ONPG (100%) and moderate activity for its natural substrate lactose (25.7%). However, the hydrolysis ability of the enzyme towards all other chromogenic nitrophenyl analogues is very weak, indicating that Gal308 is a β-galactosidase with narrow substrate specificity. To investigate the kinetic parameters of recombinant enzyme, the Michaelis-Menten constants (Km), turnover numbers (kcat), and catalytic efficiencies (kcat/Km) of Gal308 for ONPG and lactose are determined. The kcat and Km values are 464.7±7.8 s-1 and 2.7±0.3 mM for ONPG, and 264.2±2.1 s-1 and 7.1±0.8 mM for lactose, respectively. The kcat/Km value of the enzyme for ONPG (172.1 s-1mM-1) is 4.6-fold higher than that for lactose (37.2 s-1mM-1), which clearly demonstrated that the catalytic efficiency of Gal308 for ONPG is much higher than that for lactose[1].
Kinase AssayThe β-galactosidase activity is measured using two substrates including ONPG and lactose in this study. The β-galactosidase activity for ONPG is measured by following the amount o-nitrophenol released from ONPG. The reaction mixture is composed of 100 μL of the enzyme solution and 400 μL of ONPG solution (2.5 g/L in 100 mM Tris-HCl buffer at pH 6.8). After incubation at 78°C for 15 min, the reaction is terminated by adding an equal volume of 1 M Na2CO3. The released o-nitrophenol is quantitatively determined by measuring at A405. One unit of activity is defined as the amount of enzyme needed to produce 1 μmol of o-nitrophenol per minute under the assay condition. The specific activity is expressed as units per milligram of protein. Assays for activity towards lactose are performed in the same buffer containing 100 μL of enzyme solution and 5% lactose, and the reaction is stopped by boiling for 10 min, and the concentration of glucose is determined using a glucose oxidase-peroxidase assay kit. The released glucose is quantitatively determined by measuring A492. One unit of enzyme activity is defined as the amount of activity required to release 1 μmol of glucose per minute[1].
References

[1]. Zhang X, et al. Metagenomic approach for the isolation of a thermostable β-galactosidase with high tolerance of galactose and glucose from soil samples of Turpan Basin. BMC Microbiol. 2013 Oct 24;13:237. doi: 10.1186/1471-2180-13-237.

Chemical & Physical Properties

Density1.6±0.1 g/cm3
Boiling Point572.4±50.0 °C at 760 mmHg
Melting Point186-193 °C (dec.)
Molecular FormulaC12H15NO8
Molecular Weight301.249
Flash Point300.0±30.1 °C
Exact Mass301.079773
PSA145.20000
LogP-0.78
Vapour Pressure0.0±1.7 mmHg at 25°C
Index of Refraction1.648
InChIKeyKUWPCJHYPSUOFW-YBXAARCKSA-N
SMILESO=[N+]([O-])c1ccccc1OC1OC(CO)C(O)C(O)C1O
Storage condition2~8°C

Safety Information

Personal Protective EquipmentEyeshields;Gloves;type N95 (US);type P1 (EN143) respirator filter
Hazard CodesXi
Risk PhrasesR36/37/38
Safety PhrasesS22-S24/25
RIDADRNONH for all modes of transport
WGK Germany3
HS Code29400090

Customs

HS Code29400090

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Synonyms

EINECS 206-716-1
2-Nitrophenyl β-D-galactopyranoside
T6OTJ BOR BNW& CQ DQ EQ F1Q &&β-D-Galacto Form
MFCD00063255
2-Nitrophenyl β-D-Galactopyranoside [Substrate
o-Nitrophenyl β-D-galactopyranoside
β-D-Galactopyranoside, 2-nitrophenyl
2-Nitrophenyl β-D-galactoside
2-Nitrophenyl beta-D-Galactopyranoside
2-Nitrophenyl β-D-galactopyranoside (ONPG)
2-Nitrophenyl-β-D-galactopyranoside
ortho-Nitrophenyl-β-galactoside
ONPG
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