In the intricate dance of life, enzymes play a quiet yet powerful role. Among them, creatinine amidohydrolase stands out as a specialized tool in clinical diagnostics and biochemical research. Derived from microorganisms and engineered with precision, this enzyme transforms creatinine into creatine, paving the way for accurate creatinine detection in laboratory assays.
Whether you're a researcher, clinician, or laboratory technician, understanding how this enzyme works can elevate your biochemical workflows and improve the reliability of your results.

What Is Creatinine Amidohydrolase?

Creatinine amidohydrolase (EC 3.5.2.10) is a hydrolytic enzyme that catalyzes the reaction:

Creatinine + H₂O → Creatine
This enzyme is commonly used in tandem with creatine amidohydrolase and sarcosine oxidase to enzymatically determine creatinine levels—an essential marker for kidney function in clinical diagnostics. The recombinant form of this enzyme, derived from microorganisms, ensures high purity and consistent performance in lab settings.

Key Properties of Creatinine Amidohydrolase
When working with enzymes, consistency and stability are everything. Here’s what makes this enzyme a reliable choice:
• Appearance: White to slightly yellowish lyophilized powder.
• Purity: ≥90% (SDS-PAGE)
• Specific Activity: ≥400 U/mg
• Molecular Weight: 29 kDa
• Isoelectric Point (pI): 5.3
• Optimum pH: 7.0–8.0
• Optimum Temperature: 65°C
• Storage Stability: At least one year at -25°C to -15°C

• Inhibitors: Sensitive to Hg²⁺, Cu²⁺, and Fe³⁺

This remarkable stability across a broad pH range (5.5–10.0) and temperature tolerance makes it a robust performer in most lab environments.

Applications in Clinical Diagnostics
Creatinine amidohydrolase plays a central role in enzymatic creatinine assays, helping laboratories assess renal health quickly and accurately. When combined with downstream enzymes, it allows the detection of creatinine via colorimetric methods—often using spectrophotometry at 525 nm.
These methods are commonly used in:
• Routine kidney function testing
• Automated clinical analyzers

• Research studies involving renal biomarkers

By ensuring consistent conversion of creatinine to creatine, this enzyme supports reliable and reproducible results in healthcare diagnostics.

How to Use Creatinine Amidohydrolase in Enzyme Assays
Using creatinine amidohydrolase correctly is essential for precise measurements. Here is a simplified step-by-step workflow:
1.Buffer Preparation: Prepare potassium phosphate buffer (pH 6.5).
2.Enzyme Reaction: Mix creatinine solution with enzyme at 37°C.
3.Stop Reaction: Use sodium carbonate to terminate after 10 minutes.
4.Color Development: Add α-naphthol, alkaline, and diacetyl solutions.

5.Measurement: Measure absorbance at 525 nm.

This spectrophotometric method quantifies creatine formation, indicating enzyme activity.

Assay Principle and Calculation

The enzyme’s activity is defined as the amount that generates 1 μmol of creatine per minute. Using a specific extinction coefficient (0.0704 cm²/μmol), labs can calculate precise enzyme activity for quality control or diagnostic interpretation.

Formula:
Enzyme activity (U/mg) = (ΔA × df × 1.0) / (0.0704 × 0.1 × 10 × C)
Where:
• ΔA = Absorbance difference
• df = Dilution factor
• C = Enzyme concentration (mg/mL)

Why Choose HH0106 Creatinine Amidohydrolase?
The HH0106 series of creatinine amidohydrolase offers unmatched purity and performance. With minimal contamination from other hydrolases and ATPases, this enzyme ensures clean and specific reactions. It is stabilized with BSA and sugar, enhancing shelf-life and reliability during shipping and storage.

Conclusion: Nature and Science in Harmony
Creatinine amidohydrolase is a perfect example of how biotechnology and nature collaborate to serve human health. This enzyme not only brings precision to kidney diagnostics but also embodies the beauty of molecular design—working quietly, faithfully, and effectively.
By choosing a high-quality creatinine amidohydrolase like HH0106, labs can achieve dependable results and contribute to better patient care through accurate diagnostics.