ONPG (2-Nitrophenyl β-D-galactopyranoside) (Synonyms: ortho-Nitrophenyl β-D-galactopyranoside, o-Nitrophenyl β-D-Galactoside, ortho-Nitrophenyl β-D-Galactoside, NSC 83631, ONPG) |
Catalog No.GC33431 |
ONPG (2-Nitrophenyl β-D-galactopyranoside) is a colorimetric and spectrophotometric substrate for detection of β-galactosidase activity.
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Cas No.: 369-07-3
Sample solution is provided at 25 µL, 10mM.
ONPG is a colorimetric and spectrophotometric substrate for detection of β-galactosidase activity.
The 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].
[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.
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