PNPG (4-Nitrophenyl β-D-glucopyranoside)
(Synonyms: 4-硝基苯-Β-D-吡喃葡萄糖苷,4-Nitrophenyl β-D-glucopyranoside) 目录号 : GC30094
A chromogenic β-glucosidase substrate
Cas No.:2492-87-7
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
4-Nitrophenyl β-D-glucopyranoside is a chromogenic substrate for β-glucosidase.1 Upon hydrolysis by β-glucosidase, 4-nitrophenol is released, which can be quantified by colorimetric detection at 405 nm as a measure of β-glucosidase activity.
1.Britton, J., Raston, C.L., and Weiss, G.A.Rapid protein immobilization for thin film continuous flow biocatalysisChem. Commun. (Camb.)52(66)10159-10162(2016)
| Cas No. | 2492-87-7 | SDF | |
| 别名 | 4-硝基苯-Β-D-吡喃葡萄糖苷,4-Nitrophenyl β-D-glucopyranoside | ||
| Canonical SMILES | O[C@H]1[C@@H](O[C@H](CO)[C@@H](O)[C@@H]1O)OC2=CC=C([N+]([O-])=O)C=C2 | ||
| 分子式 | C12H15NO8 | 分子量 | 301.25 |
| 溶解度 | DMSO : ≥ 48 mg/mL (159.34 mM) | 储存条件 | Store at -20°C, protect from light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.3195 mL | 16.5975 mL | 33.195 mL |
| 5 mM | 0.6639 mL | 3.3195 mL | 6.639 mL |
| 10 mM | 0.332 mL | 1.6598 mL | 3.3195 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Effect of a glassy gellan/polydextrose matrix on the activity of α-D-glucosidase
Carbohydr Polym 2013 Jun 5;95(1):389-96.23618284 10.1016/j.carbpol.2013.03.022
An investigation of the ability of the enzyme α-D-glucosidase to act on the substrate 4-nitrophenyl α-D-glucopyranoside (PNPG) while embedded in glassy carbohydrate matrices (deacylated gellan with polydextrose and polydextrose alone) is presented. Physicochemical characterisation of the matrices was achieved using the techniques of modulated differential scanning calorimetry, small deformation dynamic oscillation on shear, Fourier transform infra-red spectroscopy, wide angle X-ray diffraction and scanning electron microscopy. A UV-vis spectrophotometric procedure was adapted for the analysis of the activity of α-D-glucosidase in hydrolysing PNPG in the condensed carbohydrate systems. In order to derive a relationship between the structural properties of the matrix and the enzymatic activity, mechanical spectra were recorded using the combined framework of the Williams, Landel and Ferry equation with the time-temperature superposition principle. Theoretical modelling and experimental observations strongly argue for a pronounced effect of the gelling polysaccharide/co-solute mixture on enzymatic activity near the mechanical Tg of the matrix.
Effects of intron retention on properties of β-glucosidase in Aspergillus niger
Fungal Biol 2019 Jun;123(6):465-470.31126423 10.1016/j.funbio.2019.04.002
Intron retention, one of the major types of alternative splicing in plants and animals, has also been reported existing in filamentous fungi's glycoside hydrolases. In this study, an intron-retained β-glucosidase gene transcript (bgl1B) from A. niger B2 strain was obtained. Compared with the normally spliced transcript bgl1A, bgl1B had an extra 51bp insertion, which was confirmed to be the sixth (the last) intron of this β-glucosidase gene. The bgl1A and bgl1B were expressed in Pichia pastoris and the purified enzymes were used to compare their catalytic properties. The results showed that the intron retention didn't impair the catalytic function. Instead, the intron-retained enzyme BGL1B had a better thermostability with a higher optimal temperature and a longer half-life under 50 °C. Also it exhibited a little higher kcat for 4-nitrophenyl-β-d-glucopyranoside (PNPG) and a noticeable higher hydrolysis efficiency towards geniposide. This work suggested that the β-glucosidase gene in A. niger most likely underwent an alternative splicing presented as intron retention type, and intron retention might be a source of enzyme diversity in fungi.
BadGluc, a β-glucosidase from Bjerkandera adusta with anthocyanase properties
Bioprocess Biosyst Eng 2018 Sep;41(9):1391-1401.29948211 10.1007/s00449-018-1966-4
A glycosidase of the basidiomycete Bjerkandera adusta (BadGluc) was found in screenings to possess a strong decolorizing ability towards malvidin-3-galactoside, an anthocyanin abundant in various berry fruits. The BadGluc was purified from the culture supernatant via FPLC, and the corresponding gene was identified which showed low similarity to other characterized glucosidases. Scanning the primary sequence with PROSITE no active site motif was detected. Eventually, a specific 18 aa consensus pattern was identified manually. The active site motif possessed an undescribed sequence which was only found in a few hypothetical proteins. The corresponding gene was cloned and expressed in Pichia pastoris GS115 yielding activities up to 100 U/L using 4-nitrophenyl-β-d-glucopyranoside (PNPG) as substrate. The enzyme possessed a good temperature (70% after 1 h at 50°C) and pH stability (70% between pH 2 and 7.5), and preferably catalysed the hydrolysis of delphinidin-3-glucoside and cyanidin-3-glucoside, regardless of the position of the terminal Hexa-His tag. This novel glucosidase worked in aqueous solution as well as on pre-stained fabrics making it the first known candidate anthocyanase for applications in the detergent and food industries.
Highly Efficient Biotransformation of Phenolic Glycosides Using a Recombinant β - Glucosidase From White Rot Fungus Trametes trogii
Front Microbiol 2022 May 18;13:762502.35663869 PMC9158485
Phenolic glycosides are the important bioactive molecules, and their bioavailability can be influenced by enzyme hydrolysis, such as β-glucosidases (EC3.2.1.21) and other glycosyl hydrolases (GHs). Wood rotting fungi possess a superfamily of GHs, but little attention has been paid to the GHs and their potential applications in biotransformation of phenolic glycosides. In this study, two GH3 gene family members of Trametes trogii S0301, mainly expressed in the carbon sources conversion stage were cloned, and TtBgl3 coded by T_trogii_12914 showed β-glucosidase activity toward 4-nitrophenyl β-D-glucopyranoside (PNPG). The recombinant TtBgl3 preferred an intermediately neutral optimum pH with >80% of the maximum activity at pH 5.0-7.0 and was stable at a wide range of pH (5.0-10.0). Phenolic glycosides transformation experiments showed that TtBgl3 was a dual-activity enzyme with both activities of aryl-β-D-glucosidase and β-glucuronidase, and could hydrolyze the β-glucoside/glucuronide bond of phenolic glycosides. Under optimized conditions, the recombinant TtBgl3 had much higher transformation efficiency toward the β-glucoside bond of gastrodin, esculin and daidzin than β-glucuronide bond of baicalin, with the transformation rate of 100 and 50%, respectively. Our homology modeling, molecular docking, and mutational analysis demonstrated that His85 and Lys467 in the acceptor-binding pocket of TtBgl3 were the potential active sites. The point mutation of His85 and Lys467 leads to the significantly impaired catalytic activity toward PNPG and also the weak transformation efficiency toward gastrodin. These findings provide insights for the identification of novel GH3 β-glucosidases from T. trogii and other wood-rotting fungi. Furthermore, TtBgl3 might be applied as green and efficient biological catalysts in the deglycosylation of diverse phenolics to produce bioactive glycosides for drug discovery in the future.
Structural and biochemical characterization of a GH3 β-glucosidase from the probiotic bacteria Bifidobacterium adolescentis
Biochimie 2018 May;148:107-115.29555372 10.1016/j.biochi.2018.03.007
Bifidobacterium is an important genus of probiotic bacteria colonizing the human gut. These bacteria can uptake oligosaccharides for the fermentative metabolism of hexoses and pentoses, producing lactate, acetate as well as short-chain fatty acids and propionate. These end-products are known to have important effects on human health. β-glucosidases (EC 3.2.1.21) are pivotal enzymes for the metabolism and homeostasis of Bifidobacterium, since they hydrolyze small and soluble saccharides, typically producing glucose. Here we describe the cloning, expression, biochemical characterization and the first X-ray structure of a GH3 β-glucosidase from the probiotic bacteria Bifidobacterium adolescentis (BaBgl3). The purified BaBgl3 showed a maximal activity at 45 °C and pH 6.5. Under the optimum conditions, BaBgl3 is highly active on 4-nitrophenyl-β-d-glucopyranoside (PNPG) and, at a lesser degree, on 4-nitrophenyl-β-d-xylopyranoside (pNPX, about 32% of the activity observed for PNPG). The 2.4 Å resolution crystal structure of BaBgl3 revealed a three-domain structure composed of a TIM barrel domain, which together with α/β sandwich domain accommodate the active site and a third C-terminal fibronectin type III (FnIII) domain with unknown function. Modeling of the substrate in the active site indicates that an aspartate interacts with the hydroxyl group of the C6 present in PNPG but absent in pNPX, which explains the substrate preference. Finally, the enzyme is significantly stabilized by glycerol and galactose, resulting in considerable increase in the enzyme activity and its lifetime. The structural and biochemical studies presented here provide a deeper understanding of the molecular mechanisms of complex carbohydrates degradation utilized by probiotic bacteria as well as for the development of new prebiotic oligosaccharides.