Chlorophenol red-b-D-galactopyranoside sodium salt, CPRG

Chlorophenol red-b-D-galactopyranoside sodium salt is a fluorescent dye that belongs to the group of phenolic compounds. It is soluble in water and organic solvents. Chlorophenol red-b-D-galactopyranoside sodium salt has been used as a light emission agent for tissue culture and has been shown to have genotoxic activity in some studies, but not others. This compound can be used as a fluorescence probe for biological studies and structural analysis of proteins, nucleic acids, and lipids. It also has been found to have protease activity in vitro. Chlorophenol red-b-D-galactopyranoside sodium salt is also toxic to cells in culture at high concentrations and may cause cell lysis and clinical relevance.

Chlorophenol red beta-D-galactopyranoside sodium salt (CPRG) is a chemical compound used in scientific experiments to detect the presence of beta-galactosidase, an enzyme commonly found in bacteria. CPRG produces a red color when cleaved by beta-galactosidase, which makes it a useful tool in microbiological research.

Definition and Background:

CPRG is a synthetic substrate used to detect beta-galactosidase activity. Beta-galactosidase is an enzyme that cleaves lactose into glucose and galactose, and is found in many bacteria, including E. coli. CPRG is commonly used in the production of recombinant proteins to monitor gene expression and assess the efficacy of gene therapy.

Physical and Chemical Properties:

CPRG is a red-orange powder that is soluble in water and ethanol. It has a molecular weight of 524.5 and a melting point of 245-250°C. Its chemical formula is C18H17CINO8SNa, and its IUPAC name is sodium (4-chloro-3-[(4-methoxybenzoyl)amino]phenyl)(6-oxo-6-(2-thienylamino) hexylidene) azanide.

Synthesis and Characterization:

CPRG is synthesized by reacting 4-chloro-3-nitrobenzoyl chloride with 6-(2-thienylamino)hexylamine, followed by reaction with sodium 4-(dimethylamino)phenolate in the presence of sodium methoxide. The product is then purified by recrystallization.

Analytical Methods:

CPRG is commonly analyzed using UV-Vis spectrophotometry, which measures the absorbance of the color produced by cleavage with beta-galactosidase. Other methods include high-performance liquid chromatography (HPLC), mass spectrometry, and NMR spectroscopy.

Biological Properties:

CPRG is non-toxic and non-hazardous to human health. It has no reported mutagenic, teratogenic, or carcinogenic properties. It is also biodegradable, making it an environmentally friendly alternative to other substrates used to detect beta-galactosidase.

Toxicity and Safety in Scientific Experiments:

CPRG is safe for use in scientific experiments when handled and disposed of properly. It is important to wear gloves and protective clothing when handling the powder, as it may irritate the skin and eyes.

Applications in Scientific Experiments:

CPRG is used to detect beta-galactosidase activity in bacterial cultures. This is useful in measuring gene expression, assessing the effectiveness of gene therapy, and in identifying bacterial species. CPRG can also be used in the production of recombinant proteins, as it allows researchers to monitor the expression of the protein in real-time.

Current State of Research:

Research on CPRG is ongoing, with a focus on developing more sensitive and efficient detection methods. There is also an interest in developing new substrates for detecting other enzymes commonly found in bacteria, such as alkaline phosphatase and beta-lactamase.

Potential Implications in Various Fields of Research and Industry:

CPRG has potential applications in a variety of fields, including microbiology, genetic engineering, biotechnology, and pharmaceuticals. Its ability to detect gene expression and protein production makes it a valuable tool in developing new treatments for diseases and in studying the mechanisms of disease.

Limitations and Future Directions:

Despite its many benefits, CPRG does have some limitations. It is limited to detecting beta-galactosidase activity in bacterial cultures and cannot be used to detect the activity of other enzymes or in other organisms. Future research will likely focus on developing new substrates to detect a wider range of enzymes, and on improving the sensitivity and efficiency of detection methods.

Future Directions:

1. Developing new substrates to detect other enzymes commonly found in bacteria.

2. Improving the sensitivity and efficiency of detection methods.

3. Expanding the use of CPRG to non-bacterial organisms.

4. Developing new applications for CPRG in areas such as food safety and environmental monitoring.

5. Studying the mechanisms of beta-galactosidase activity in bacteria.

6. Developing new treatments for diseases based on the mechanisms of beta-galactosidase activity.

7. Developing new techniques for gene therapy using CPRG.

8. Optimizing the production and purification of recombinant proteins using CPRG.

9. Developing new methods for analyzing the data generated by CPRG assays.

10. Studying the toxicity and environmental impact of CPRG and other substrates used to detect enzyme activity.