Enzyme Substrates

Fluorescein-dipalmitoylphosphatidylethanolamine triethylammonium salt is a fluorescent phospholipid derivative for phospholipase. This substrate consists of a fluorescein conjugated to DPPE (dipalmitoylphosphatidylethanolamine), a key component of biological membranes. This compound is frequently used in membrane dynamics studies and lipid metabolism research. The fluorescein moiety allows visualization of lipid interactions in model membranes, liposomes, or live-cell imaging applications. The triethylammonium salt form enhances solubility and stability, property that facilitates its use in aqueous and lipid-based environments.

Aldol® 518 myo-inositol phosphate is a chromogenic and fluorogenic enzyme substrate used for detection of phosphatidylinositol specific phospholipase C (PI-PLC, EC 4.6.1.13) activity. The colorless enzyme substrate is cleaved by PI-PLC positive microorganisms grown in liquid or solid media, yielding intense red color. In combination with Aldol® 355 fluorescence enhancer, red fluorescence can also be obtained. Aldol® 518 myo-inositol phosphate thus enables the detection and identification of various bacteria, such as, pathogenic strains of Listeria, Clostridium, Bacillus, and Staphylococcus species. Aldol® 518 myo-inositol phosphate is suitable for use under aerobic and anaerobic conditions.

Aldol® 515 N-glutaryl-L-phenylalanine amide is a chromogenic and fluorogenic enzyme substrate used for detection of L-glutamate-phenylalanine (GluPhe)-specific peptidase activity. The colorless enzyme substrate is cleaved the GluPhe-specific peptidase, yielding development of orange-red coloration. In addition, red fluorescence is generated when Aldol® 355 fluorescence enhancer or a suitable matrix is present. Aldol® 515 N-glutaryl-L-phenylalanine amide can be used in enzyme assays and live bacterial cultures. Aldol® 515 N-glutaryl-L-phenylalanine amide is suitable for use in liquid and solid media, under aerobic as well as anaerobic conditions.

Chromogenic substrate for esterase with C9 activity yielding a salmon colored precipitate upon cleavage.

2,4-Dinitrophenyl 2-deoxy-2-fluoro-beta-xylobioside, also known as DNFPX, is a substrate to study the activity of glycosidases. It is a derivative of xylobiose, which is a disaccharide composed of two xylose units. The 2-deoxy-2-fluoro modification on one of the xylose units makes the compound resistant to further hydrolysis by the enzyme, allowing it to be used as a reporter of enzyme activity. The 2,4-Dinitrophenyl group is used as a chromogenic group that enables rapid detection and measurement of the enzyme activity that can be quantitatively measured by monitoring the appearance of the yellow 2,4-dinitrophenyl group when cleavage of the glycosidic bond.

4-Methylumbelliferyl N-acetyl-alpha-D-neuraminic acid ammonium is a fluorogenic substrate for neuraminidase. After enzymatic cleaveage, free 4-methylumbelliferone (also known as hymecromone) is released, exhibiting blue fluorescence upon excitation with UV light. The strongest fluorescence of 4-methylumbelliferone requires deprotonation of the hydroxyl group (thus requires alkaline pH), with a maximal fluorescence intensity obtained with excitation at 350 to 370 nm and emission at 440 to 470 nm. The use of 4-methylumbelliferyl N-acetyl-alpha-D-neuraminic acid ammoniumas a substrate for measuring the neuraminidase activity is used for detecting the glycoproteins on the surface of influenza A and B viruses, as well as for sialidosis diagnosis.

2-Nitrophenyl 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-a-D-glucopyranoside is a chromogenic substrate commonly used in enzyme assays for the detection and quantification of enzyme activity. It is a colorless substrate that is enzymatically converted into a yellowish product, the intensity of which is directly proportional to the enzyme activity present in the sample.

4-Methylumbelliferyl N,N-diacetyl-b-D-chitobioside is a fluorogenic substrate for peptide hormones. It is used in vitro to study the polymerase chain reaction (PCR) of human serum and other biological samples. 4-Methylumbelliferyl N,N-diacetyl-b-D-chitobioside is an orally bioavailable prodrug that undergoes hydrolysis by esterases to release 4-methylumbelliferone, which is then converted into the fluorescent product methylumbelliferone by cellular esterases. This product can be detected using a fluorescence sensor with high specificity and sensitivity. The fluorescence signal generated by this product is proportional to the enzyme concentration and can be used as a biomarker for peptide hormone production or neuronal death.

Histochemical substrate for alkaline phosphatase

Applications A histochemical substrate for a-N-acetyl-galactosaminidase. References Zhu, et al.: Gene, 137, 309 (1994)

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3-Carboxyumbelliferyl b-D-galactoside BSA conjugate is a fluorogenic substrate that is used in chemiluminescent enzyme assays. The reaction of this compound with the enzyme β-galactosidase leads to an increase in the intensity of light emitted by the compound, which can be measured using a luminometer. This product is often used in diagnostics, such as food testing and environmental testing, as well as high quality research.

Chromogenic substrate for beta-glucosidase

Chromogenic substrate for β-glucuronidase (GUS) yieldinga blue precipitate. It is used for detection of GUS+ bacterial colonies and has been reported as chromogenic substrate in a number of chromogenic E. coli / coliform culture media. About 95% of common E. coli possess β-D-glucuronidase activity. Such chromogenic culture media are used for detection and enumeration of E. coli in food samples such as meat, dairy products, shellfish and have clinical applications in the assessment of urinary infections. It has gained international acceptance as highly accurate test for the presence of E. coli in drinking water samples. It is also used for the detection of the expression of GUS (E. coli β-glucuronidase) gene fusion marker constructs in plants.