2-Nitrophenyl b-D-glucopyranoside

2-Nitrophenyl beta-D-glucopyranoside (2NPG) is a compound that has been used extensively in scientific research. It is a chromogenic substrate for the assay of β-glucosidase, an enzyme that cleaves glycosidic bonds in carbohydrates. Its use has become increasingly popular in recent years, and this paper analyzes its definition and background, physical and chemical properties, synthesis and characterization, analytical methods, biological properties, toxicity and safety in scientific experiments, applications in scientific experiments, and potential implications in various fields of research and industry.

Definition and Background

2NPG is a colorless crystalline compound that is soluble in water, chloroform, methanol, and other polar solvents. It has a molecular weight of 331.28 g/mol, and its chemical structure is shown as below:

2NPG is a chromogenic substrate that releases p-nitrophenol (pNP) upon hydrolysis by β-glucosidase. It is a commonly used compound for the detection and quantification of β-glucosidase in various organisms, ranging from bacteria to plants.

Physical and Chemical Properties

2NPG is a stable compound that is not affected by light or heat. It has a melting point of 159-162 °C and a boiling point of 516.91 °C at 760 mmHg. The compound is sensitive to pH and is known to decompose rapidly in acidic conditions. Its solubility in water is 10-25 g/L, and it has a logP of 0.12, indicating that it is a hydrophilic compound.

Synthesis and Characterization

2NPG can be synthesized by reacting 2-nitrophenol with β-D-glucopyranosyl bromide in the presence of a base catalyst such as triethylamine. The reaction produces 2NPG as the primary product, with minor amounts of by-products such as glucose and p-nitrophenyl alpha-D-glucopyranoside.

The compound is often characterized using techniques such as nuclear magnetic resonance (NMR) spectroscopy, high-performance liquid chromatography (HPLC), and mass spectrometry (MS). The NMR spectrum of 2NPG shows characteristic peaks for the hydroxyprotons of glucose and the aromatic protons of nitrophenyl. HPLC and MS can be used to determine the purity of the compound and quantify the amount of 2NPG in a sample.

Analytical Methods

2NPG is commonly used as a substrate for the detection and quantification of β-glucosidase activity. The hydrolysis of 2NPG by β-glucosidase produces pNP, which absorbs light at 405 nm. The amount of pNP released can be measured using a spectrophotometer, and the enzyme activity can be calculated based on the rate of the reaction.

Other analytical methods that have been used to study 2NPG include electrochemical methods, fluorescence assays, and capillary electrophoresis. Electrochemical methods involve measuring the current or potential changes produced by the cleavage of 2NPG by an enzyme. Fluorescence assays use fluorescently labeled β-glucosidases or 2NPG analogs to measure enzyme activity. Capillary electrophoresis separates and detects 2NPG and its hydrolysis products based on their electrophoretic mobility.

Biological Properties

2NPG is a versatile compound that has been used to assay β-glucosidase activity in various organisms. It has been used to study the effects of temperature, pH, salt concentration, and other environmental factors on the enzyme activity. It has also been used to identify and characterize new β-glucosidase enzymes.

In addition to its use as a chromogenic substrate, 2NPG has been shown to have other biological activities. It has been reported to have antibacterial activity against some gram-positive and gram-negative bacteria. It has also been shown to inhibit the growth of certain cancer cells and induce apoptosis in human leukemia cells.

Toxicity and Safety in Scientific Experiments

The toxicity of 2NPG has been studied in various organisms, including humans, animals, and microorganisms. In general, it has low toxicity and is considered safe to use in scientific experiments.

However, some studies have reported adverse effects of 2NPG at high concentrations. For example, exposure to high concentrations of 2NPG has been shown to cause oxidative stress and DNA damage in human liver cells. It has also been shown to inhibit the growth of some beneficial bacteria in the gut, leading to dysbiosis.

Applications in Scientific Experiments

2NPG has numerous applications in scientific research. It is a widely used compound for the assay of β-glucosidase activity in various organisms. It has also been used to characterize and identify new β-glucosidase enzymes.

In addition, 2NPG has been used as a substrate for the synthesis of other compounds. For example, it has been used as a starting material for the synthesis of glycosides, which have antibacterial, antiviral, and antifungal activities. It has also been used to synthesize compounds with potential anticancer activities.

Current State of Research

The use of 2NPG in scientific research continues to expand. New analytical methods and applications for 2NPG are being developed, and the compound is increasingly being used to study β-glucosidase enzymes in diverse organisms.

Potential Implications in Various Fields of Research and Industry

The use of 2NPG has potential implications in various fields of research and industry. It has applications in biotechnology, pharmaceuticals, and agriculture. For example, it can be used to develop new β-glucosidase enzymes for the production of biofuels from lignocellulosic biomass. It can also be used to develop new antibiotics and other drugs.

Limitations and Future Directions

Despite its many uses, 2NPG also has some limitations. For example, it is sensitive to pH and can decompose rapidly in acidic conditions. It can also interfere with some analytical methods, such as colorimetric assays for the detection of pNP.

Future directions for research on 2NPG include developing new analytical methods and applications for the compound. For example, new electrochemical and fluorescence-based assays can be developed to improve the sensitivity and specificity of β-glucosidase detection. 2NPG can also be used to study the effects of β-glucosidase activity on the environment and human health.

In conclusion, 2NPG is a valuable compound for the study of β-glucosidase enzymes and has numerous applications in scientific research. Its physical and chemical properties, synthesis and characterization, analytical methods, and biological properties have been extensively studied. The future directions for research on 2NPG include developing new analytical methods and applications for the compound to improve the sensitivity and specificity of β-glucosidase detection and to study its effects on the environment and human health.

COA:

Product name: 2'-Nitrophenyl-beta-D-glucopyranoside

CAS: 2816-24-2     M.F.: C₁₂H₁₅NO₈          M.W._: 301.25