2-Nitrophenyl 2,3,4-tri-O-acetyl-b-D-xylopyranoside

2-Nitrophenyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside is a molecule that has been shown to be clastogenic in the micronucleus test. It is also mutagenic and teratogenic in the L5178Y mouse lymphoma cell assay. This compound disrupts metabolic pathways by inhibiting the enzyme aspartate transaminase, which catalyzes the conversion of aspartic acid to oxaloacetic acid. This inhibition leads to an accumulation of oxaloacetate and lactic acid. The mechanism of action is similar to that of phenylbutazone, which is a nonsteroidal anti-inflammatory drug (NSAID) that inhibits cyclooxygenase enzymes. 2NPTXR can be used as a fluorescent probe for DNA replication in vitro and in vivo.

2-Nitrophenyl 2,3,4-tri-O-acetyl-b-D-xylopyranoside, commonly known as NPX, is a chemical compound of the nitrophenoxypentyl glycosides family. The chemical structure of NPX depicts the presence of a nitrophenyl moiety, xylopyranoside, and three acetyl groups. It is widely used in the field of glycomics to determine the activity of glycoside hydrolases, enzymes that degrade glycosidic bonds. The following sections provide an overview of the physical and chemical properties, synthesis, characterization, biological properties, and potential implications of NPX in various fields of research and industry.

Synthesis and Characterization

NPX can be synthesized by the reaction of 2-nitrophenol with 2,3,4-tri-O-acetyl-β-D-xylopyranosyl bromide in the presence of a base, such as potassium carbonate, and an organic solvent, such as acetone. The reaction proceeds via a substitution mechanism, in which the bromide group is replaced by the nitrophenyl group. The acetyl groups protect the xylopyranosyl moiety from unwanted reactions and can be removed by acidic or basic hydrolysis.

The characterization of NPX can be done by various analytical techniques, such as nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS), and high-performance liquid chromatography (HPLC). The NMR spectrum of NPX shows distinct peaks for the xylopyranosyl moiety and the acetyl groups, which can be used to determine the purity and identity of the compound. The MS spectrum of NPX shows the molecular ion peak at m/z = 427, which confirms the molecular weight of the compound. The HPLC analysis of NPX can be used to determine the retention time, which is dependent on the polarity and composition of the mobile and stationary phases.

Analytical Methods

NPX is widely used as a substrate to measure the activity of glycoside hydrolases, which are enzymes that cleave glycosidic bonds between monosaccharides. The cleavage of NPX by glycoside hydrolases results in the release of the aglycone, 2-nitrophenol, which can be detected spectrophotometrically at 400 nm. The rate of the reaction is dependent on the concentration of the enzyme, the pH, the temperature, and the substrate concentration. The Michaelis-Menten kinetics can be used to determine the parameters of the reaction, such as the Km and Vmax values.

Biological Properties

NPX has been shown to be a useful tool to monitor the activity of glycoside hydrolases in various organisms, such as bacteria, fungi, plants, and animals. The specificity of NPX towards different glycoside hydrolases can be modulated by varying the substituents on the xylopyranosyl moiety. For example, the substitution of xylopyranosyl with mannose or glucose can result in more specific substrates for certain glycoside hydrolases.

Toxicity and Safety in Scientific Experiments

NPX is considered to be of low toxicity and is not classified as a hazardous material. However, it should be handled with care and appropriate protective measures should be taken to avoid contact with skin and eyes. The disposal of NPX should be done in accordance with the local regulations for hazardous waste management.

Applications in Scientific Experiments

NPX has been extensively used in the field of glycomics to study the activity, specificity, and inhibition of glycoside hydrolases. It has also been used as a tool to study the biosynthesis and degradation of glycans in various organisms. NPX can be modified with various tags, such as fluorescent and radioactive groups, to enable the detection and quantification of glycoside hydrolase activity in vitro and in vivo.

Current State of Research

The research on NPX has mainly focused on the optimization of the synthesis and characterization of the compound, the development of new analytical methods for glycoside hydrolase activity measurement, and the application of NPX in various fields of research, such as enzymology, biochemistry, microbiology, and drug discovery.

Potential Implications in Various Fields of Research and Industry

NPX has potential implications in various fields of research and industry, such as:

• Glycomics: NPX can be used as a valuable tool to study the structure and function of glycans and glycoside hydrolases in various organisms. This can lead to the development of novel therapies for diseases that involve glycan dysregulation, such as cancer and inflammation.

• Biotechnology: NPX can be used as a substrate for the screening and characterization of glycoside hydrolases from environmental and industrial samples. This can lead to the identification and optimization of enzymes that can be used for biocatalysis and bioconversion of carbohydrates.

• Pharmacology: NPX can be used as a probe to study the metabolism and pharmacokinetics of drugs that contain glycosidic bonds. This can lead to the optimization of drug design and formulation for improved efficacy and safety.

Limitations and Future Directions

Despite its usefulness, NPX has some limitations that need to be addressed in future research. These include:

• Specificity: NPX has limited specificity towards certain glycoside hydrolases, which can result in false positives or false negatives in enzymatic assays. This can be overcome by using more specific substrates or developing new analytical methods that can distinguish between closely related enzymes.

• Stability: NPX is known to be unstable under acidic and basic conditions, which can result in the degradation of the compound and the loss of enzymatic activity. This can be overcome by modifying the substituents on the xylopyranosyl moiety or by developing more stable analogs of NPX.

• Standardization: NPX lacks a standard reference material that can be used for interlaboratory comparison and validation of glycoside hydrolase activity measurements. This can be overcome by developing a certified reference material that can be used for quality control and assurance in glycomics research.

Future directions for research on NPX include:

• Development of new synthetic routes for NPX and its analogs that are more efficient, scalable and cost-effective.

• Optimization of analytical methods for glycoside hydrolase activity measurements, such as enzyme-linked immunosorbent assay (ELISA), capillary electrophoresis (CE), and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS).

• Application of NPX in new fields of research, such as glycoengineering, glycan biomarker discovery, and glycan-mediated host-pathogen interactions.

• Integration of NPX with other glycan tools, such as lectins, antibodies, and enzymes, to enable more comprehensive and accurate analysis of glycan structures and functions.