Description

Prolyl-4-hydroxylase (EC 1.14.11.2) is an essential member of the dioxygenase enzyme family. Using α-ketoglutarate and O₂ as cosubstrates, and relying on Fe²⁺ as a cofactor, it catalyzes the formation of trans-4-hydroxy-L-proline (L-Hyp). Our prolyl-4-hydroxylase is expressed in Escherichia coli and purified using affinity chromatography. It has a molecular weight of 29.7kDa, contains a 6× His-tag, and performs optimally at pH 6.0–8.0 and 37°C. Its activity is inhibited by divalent cations, proline analogs, pyridine 2,4(5)-dicarboxylates, 3,4-dihydroxybenzoates, β-lactam antibiotics, and anthracycline compounds.

Applications

• Industrial Production: Catalyzes L-proline to L-hydroxyproline, applicable in food, pharmaceutical, and cosmetic industries.
• Medical Aesthetics: Collagen, an extracellular matrix protein, constitutes up to three-quarters of the dry weight in skin tissues. Prolyl-4-hydroxylase catalyzes proline into varying levels of hydroxyproline in mature collagen, stabilizing its triple-helix structure and enhancing tensile strength. Recombinant prolyl-4-hydroxylase can serve as a bioactive ingredient, reducing fine lines and restoring skin elasticity and smoothness.

Quality Parameters

• Appearance: White or off-white lyophilized powder
• Purity (SDS-PAGE): ≥95% 
• Relative Molecular Weight: 29.7kDa 
• Specific Activity: ≥31.0 U/mg 
• Endotoxin: <10 EU/mg

Notes: 

① CoA indicators can be customized upon request to include additional tests like microbial limits, host protein residues, host DNA residues, and heavy metal residues. 

② Enzyme activity definition: One unit of activity is defined as the enzyme amount required to catalyze the generation of 1μmol of L-Hyp per minute at 36±2°C in a MES pH 6.5 buffer containing 12mmol/L L-proline, 24mmol/L α-ketoglutarate, 1mmol/L ferrous sulfate, and 8mmol/L ascorbic acid.

Advantages

• Manufactured under GMP-compliant conditions with a batch production scale exceeding 100g, ensuring consistent quality across production batches. No animal-derived materials are used in the process.
• Low molecular weight, enabling faster absorption in localized skin applications through PH20 (recombinant human hyaluronidase) or physical permeation methods (e.g., microneedles or ultrasound).

Storage 

Lyophilized powder is stable for 24 months when stored below -15°C. After reconstitution, it can be stored at 2–8°C for up to 2 days.