Enzyme Substrates

Aldol® 518 alpha-D-glucopyranoside is a fluorogenic enzyme substrate used for detection of alpha-D-glucosidase activity. The colorless enzyme substrate is cleaved by alpha-D-glucosidase, generating red fluorescent signal and in some conditions, an orange colour in the visible spectrum can get formed, staining bacterial colonies in faint orange coloration. Aldol® 518 alpha-D-glucopyranoside can be used under aerobic and anaerobic conditions.

Calcium 2-naphthylphosphate, which is the calcim salt of 2-naphthylphosphate, is a chromogenic substrate to measure alkaline phosphatase activity. After enzymatic cleavage, the product yielded (2-naphthol) generates a red-colored complex upon reaction with a diazonium salt like Fast Blue RR or Fast Garnet GBC.

Aldol® 515 caprylate is a chromogenic and fluorogenic enzyme substrate used for detection of lipase activity. The colorless enzyme substrate is cleaved by C8 esterases, yielding development of red coloration. In addition, red fluorescence is generated when Aldol® 355 fluorescence enhancer or a suitable matrix is present. Aldol® 515 caprylate can be used in enzyme assays and live bacterial cultures. Aldol® 515 caprylate is suitable for use in liquid and solid media, under aerobic as well as anaerobic conditions.

6-Chloro-3-indolyl butyrate is a chromogenic substrate for lipases. Use in lipid digestion and metabolism studies, it produces a salmon-pink color when 6-chloro-3-indolyl is released after enzymatic cleavage.

4-Methylumbelliferyl 3,4,6-tri-O-acetyl-2-deoxy-2-trifluoroacetamido-b-D-glucopyranoside is a fluorogenic substrate specifically designed for the detection and quantification of hexosaminidase enzymes. Upon cleavage by the target enzyme, it generates a bright fluorescent signal facilitating the investigation of enzyme activity, screening of inhibitors, and diagnostics.

For the detection of beta-galactosidase activity

Histochemical substrate for esterase

Luc-Salmonella is a bioluminescent substrate for bacterial luciferase use to detect Salmonella.

Methyl 4-nitrophenyl hexylphosphonate is a phosphonate inhibitor of the enzyme serine esterase. It has been shown to be more potent than other irreversible inhibitors such as ethyl 4-nitrophenyl hexylphosphonate and methyl 4-nitrophenoxybutyl phosphate. This compound is an immobilized form of the methyl 4-nitrophenyl hexylphosphonate that is immobilized on an inert surface, such as silica gel, via a covalent bonding process. The immobilization of the molecule ensures that it will not leach from the surface and is stable over time. The molecular weight of this substance is 411.5 Daltons and its molecular formula is C8H15NO4P. This substance has a diameter of 0.8 nm and can be found in crystalline form with a melting point of 143 degrees Celsius at atmospheric pressure. Methyl 4-nitrophenyl

Applications Substrate for TenA enzyme in the thiamin salvage pathway References Jenkins, A., et al.: Nat. Chem. Biol., 3, 492 (2007), Jenkins, A., et al.: Bioorg. Chem., 36, 29 (2008),

4-Methylumbelliferyl 2-acetamido-2-deoxy-3-O-(alpha-L-fucopyranosyl)-beta-D-glucopyranoside is a fluorogenic substrate for alpha-L-fucosidase. 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 2-acetamido-2-deoxy-3-O-(alpha-L-fucopyranosyl)-beta-D-glucopyranosideas a substrate for measuring the alpha-L-fucosidase activity is used for fucosidosis diagnosis (deficiency of the enzyme alpha-L-fucosidase) and lysosomal storage disorder screening.

2-Chloro-4-nitrophenyl alpha-D-mannopyranoside is a substrate used to measure the alpha-mannosidase activity. This enzyme plays a crucial role in the lysosomal degradation of asparagine-linked carbohydrate cores of glycoproteinsis. The use of 2-Chloro-4-nitrophenyl alpha-D-mannopyranoside in diagnostic assays is important as a deficiency in alpha-mannosidase leads to the accumulation of mannose-rich oligosaccharides in lysosomes, causing a range of health problems collectively known as alpha-mannosidosis.

Sensitive fluorogenic substrate for fluorometric and UV-spectrophotometric assays of phosphatases.

5-(Pentafluorobenzoylamino)fluoroscein di-b-D-galactopyranoside, commonly referred as PFB-FDG or PFB-FDGal, is a cell-permeable enzyme substrate for β-galactosidase used in live cell imaging assays. After cleavage, 5-(pentafluorobenzoylamino)fluorescein (PFB-F) is released, generating a green fluorescence (Em ~520 nm)) of a higher intensity than fluorescein. Its main use is for bacterial detection (e.g. E. coli LacZ assays), but also for flow cytometry analytical assays.

4-Nitrophenyl 2-acetamido-3,6-di-O-acetyl-4-O-(2,3,4,6-tetra-O-acetyl-b-D-galactopyranosyl)-2-deoxy-b-D-glucopyranoside is modified acetylated chromogenic substrate designed for the detection and quantification of enzyme activity. This highly specific substrate is cleaved by enzymes, releasing a yellow 4-nitrophenol product that can be easily measured spectrophotometrically. This allows for accurate and sensitive analysis of enzyme kinetics, inhibition studies, and substrate specificity determination.

5-Bromo-4-chloro-3-indolyl a-D-xylopyranoside is a chromogenic substrate used to detect the specific enzymatic activity of alpha-xylosidase. After cleavage, 5-bromo-4-chloro-indoxyl is released, resulting in a blue to blue-green color change in bacterial colonies or media. 5-Bromo-4-chloro-3-indolyl a-D-xylopyranoside is used in bacterial xylan metabolism studies and plant cell wall degradation assays.

2-Naphthyl-α-D-glucopyranoside is a substrate for α-glucosidase. 2-Naphthol is released upon hydrolyzation. By simultaneous coupling with a suitable staining reagent, such as hexazonium p-rosaniline, the corresponding reddish-brown azo-dye is formed. Naphthols can also be detected by fluorescence analysis.