Chinese Name: D-Alanine 7-amido-4-methylcoumarin trifluoroacetate salt

English Name: D-Alanine 7-amido-4-methylcoumarin trifluoroacetate salt

Abbreviation: H-D-Ala-AMC · TFA

CAS Number: 201847-52-1

Molecular Formula: C15H14F3N2O4

Molecular Weight: 343.28

Specifications: 1 g / 5 g / 25 g (Custom packaging supported)

Form: Off-white crystalline powder

Product Number: G04050021

Stock: Made to order

Signal: Blue fluorescence

H-D-Ala-AMC · TFA is a fluorescent protease/peptidase substrate based on AMC (7-amino-4-methylcoumarin), consisting of D-alanine (D-Ala) coupled with the AMC fluorophore via an amide bond. Its core detection logic can be summarized in three stages: "cleavage – release – fluorescence":

1. Enzyme-specific recognition and cleavage

H-D-Ala-AMC · TFA adopts the D-amino acid configuration, providing stereoselectivity for D-alanine aminopeptidase. As D-alanine is a characteristic component of bacterial cell wall peptidoglycan, this substrate offers clear specificity for bacterial enzyme activity detection. When the target enzyme is present in the sample, it specifically recognizes and hydrolyzes the D-alanine-AMC amide bond in the substrate.

2. Release of the AMC fluorophore

In the intact substrate, the AMC group is coupled with D-alanine and is in a weakly fluorescent or nearly non-fluorescent state. After enzymatic cleavage, AMC is released as free 7-amino-4-methylcoumarin molecules.

3. Production and quantification of blue fluorescence signal

Free AMC exhibits strong blue fluorescence, with an excitation peak around 341 nm and an emission peak around 441 nm. Fluorescence intensity is proportional to enzyme activity and can be monitored in real time and quantified using a standard fluorescence microplate reader.

1. Fluorescent screening of foodborne pathogens

Used to detect specific bacteria such as Enterobacteriaceae and Clostridium perfringens.

2. Rapid microbiological quality testing of chilled meat

Used to detect microbial enzyme activity collected from the surface of aerobically chilled pork and beef, allowing rapid assessment of the microbiological quality of chilled meat via fluorescence signal intensity.

3. Enzyme activity detection and enzyme kinetics studies

Used for activity determination of aminopeptidases and related peptidases, analysis of enzyme kinetic parameters (Km, Vmax), and inhibitor screening.

✅ High purity control, ensuring detection consistency

• High-purity system: Purity ≥98%, ensuring batch-to-batch stability.

• Controllable background: Reduces non-specific fluorescence interference and minimizes background impact on signal interpretation.

• Trifluoroacetate salt form: More stable than the free base, facilitating storage and use.

✅ Fluorescence detection, sensitive and efficient

• Strong fluorescence signal: Releases AMC blue fluorescence after cleavage.

• Supports quantitative analysis: Fluorescence intensity correlates with enzyme activity, enabling quantitative evaluation of enzyme activity levels.

• High detection efficiency: Based on enzyme activity fluorescence signal detection, results can be obtained in a shorter time, providing a rapid reference for food microbial screening research.

✅ Good specificity, bacteria-targeted

• D-configuration selectivity: Designed based on D-alanine structure, suitable for fluorescence detection and research of bacterial D-alanine aminopeptidase activity.

• Reduced cross-reactivity: Provides more targeted detection signals for related enzyme activity analysis in complex sample systems such as food.

✅ Platform compatible, easy to operate

• Compatible with mainstream AMC fluorescence detection platforms: Ex/Em ≈ 341/441 nm, compatible with DAPI filter sets and standard fluorescence microplate readers.

• Simple operation: "Add sample – incubate – read fluorescence" three-step procedure, no complex pre-treatment required.

• Automation-ready: Suitable for 96/384-well high-throughput screening systems.

✅ Reliable supply, cost optimization

• Cost advantage: Leveraging the domestic supply chain, the cost for the same quality is significantly lower than imported brands.

• Domestic supply: Controllable lead times and stable delivery, reducing procurement cycle risks.

• Multiple specifications available: Available in 1 g / 5 g / 25 g, with custom packaging supported.