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My CartCAS：369-07-3
Storage：Powder:-20℃，2 years;Insolvent(Mother Liquid):-20℃，6 months;-80℃，1 year
Purity：HPLC≥98%
Appearance：White to off-white Solid
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| CAS | 369-07-3 |
| Name | ONPG |
| Molecular Formula | C12H15NO8 |
| Molecular Weight | 301.25 |
| Solubility | Soluble in Water ≥10mg/mL |
| Purity | HPLC≥98% |
| Appearance | White to off-white Solid |
| Storage | Powder:-20℃，2 years;Insolvent(Mother Liquid):-20℃，6 months;-80℃，1 year |
| EC | EINECS 206-716-1 |
| MDL | MFCD00063255 |
| SMILES | O[C@H]([C@@H](O)[C@H]1O)[C@@H](O[C@@H]1CO)OC2=C([N+]([O-])=O)C=CC=C2 |
| InChIKey | KUWPCJHYPSUOFW-YBXAARCKSA-N |
| InChI | InChI=1S/C12H15NO8/c14-5-8-9(15)10(16)11(17)12(21-8)20-7-4-2-1-3-6(7)13(18)19/h1-4,8-12,14-17H,5H2/t8-,9+,10+,11-,12-/m1/s1 |
| PubChem CID | 96647 |
| Target Point | Others |
| Passage | Others |
| Background | It is used to detect β-galactosidase (β-galactosidase) activity and is a chromogenic substrate for β-galactosidase. After being catalyzed by β-galactosidase, a yellow product can be produced, and the absorbance can be detected at a wavelength of 410-420 nm. |
| Biological Activity | ONPG 是一种比色和分光光度计底物，用于检测 β-半乳糖苷酶活性。[1] |
| In Vitro | 该酶对ONPG具有高水解能力(100％)，对其天然底物乳糖具有中等活性(25.7％)。然而，酶对所有其他生色硝基苯类似物的水解能力非常弱，表明Gal308是具有窄底物特异性的β-半乳糖苷酶。为了研究重组酶的动力学参数，测定了GalP8对ONPG和乳糖的Michaelis-Menten常数(Km)，转换数(kcat)和催化效率(kcat/Km)。 ONPG的kcat和Km值分别为464.7±7.8s-1和2.7±0.3mM，乳糖分别为264.2±2.1s-1和7.1±0.8mM。 ONPG(172.1 s-1mM-1)酶的kcat/Km值比乳糖(37.2 s-1mM-1)高4.6倍，这清楚地表明Gal308对ONPG的催化效率要高得多而不是乳糖[1]。 |
| Kinase Experiment | 在该研究中使用包括ONPG和乳糖的两种底物测量β-半乳糖苷酶活性。通过遵循从ONPG释放的邻硝基苯酚的量来测量ONPG的β-半乳糖苷酶活性。反应混合物由100μL酶溶液和400μLONPG溶液(2.5g/L，在pH6.8的100mM Tris-HCl缓冲液中)组成。在78℃温育15分钟后，通过加入等体积的1M Na 2 CO 3终止反应。通过在A405测量来定量测定释放的邻硝基苯酚。一个活性单位定义为在测定条件下每分钟产生1μmol邻硝基苯酚所需的酶量。比活性表示为每毫克蛋白质的单位。在含有100μL酶溶液和5％乳糖的相同缓冲液中进行对乳糖活性的测定，并通过煮沸10分钟终止反应，并使用葡萄糖氧化酶 - 过氧化物酶测定试剂盒测定葡萄糖浓度。通过测量A492定量测定释放的葡萄糖。一个酶活性单位定义为每分钟释放1μmol葡萄糖所需的活性量[1]。 |
| Data Literature Source | [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 |
| Unit | Bottle |
| Specification | 200mg 10mM*1mL in Water 1g |
Remark：These protocols are for reference only. Solarbio does not independently validate these methods.
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