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DENARASE® High Salt - Enzyme Characteristics

Salt & Magnesium Concentration

DENARASE® High Salt has been designed to enable DNA clearance under a broad range of salt conditions. The engineered enzyme exhibits activity from 0-500 mM NaCl (see Fig. 1). Magnesium (Mg2+) is an essential cofactor for DENARASE® High Salt and minimal levels are required for basal activity (see Fig. 2).  DENARASE® High Salt is highly effective at process-relevant pH 7.4 with standard magnesium supplementation (5 mM MgCl2, see Fig. 1). The performance of the salt-adapted enzyme can be further optimized by increasing magnesium and pH levels (see Fig. 2 & 3). Optimal DENARASE® High Salt activity is observed between 5-25 mM Mg2+. However, excessive magnesium reduces the enzyme's activity (see Fig. 2).

DENARASE High Salt_Effect of NaCl

Fig. 1: Effect of NaCl, pH and magnesium

DENARASE High Salt_Effect of Magnesium-1

Fig. 2: Effect of magnesium

pH & Temperature

DENARASE High Salt_Effect of pH value in different buffer systems

Fig. 3: Effect of pH value in different buffer systems

DENARASE High Salt_Effect of Temperature

Fig. 4: Effect of temperature

Key Characteristics & Optimal Conditions

Molecular Weight (calculated):  27 kDa (per monomer)
Cofactor: Mg2+ (optimum 5 -25 mM)
pH Optimum: pH 7.4-9.0 (Fig. 3)
Temperature Optimum: 37 °C (Fig. 4)
Isolectric Point (pl, calculated): 7.83

 

Product Specification

In order to ensure a constant and high-quality level for DENARASE® High Salt, each batch must fulfill the in-house acceptance criteria for the parameters listed below.

 
Criteria Method Specification
Appearance Visual Clear, transparent solution
Activity Photometric1 > 250 U/µL
Purity Protein purity determined by
SDS-PAGE and silver staining		 ≥ 98%
Specific Activity Activity per protein content
determined photometrically
at 280 nm with a molar extinction
coefficient of 44,600 L x mol-1 x cm-1		 > 4 x 105 U/mg
Endotoxin Level LAL-Test acc. to Ph. Eur. 2.6.14/USP <85>,
Method C		 < 0.25 EU/kU
Total Microbial Count TAMC/TYMC acc. to Ph. Eur. 2.6.12/USP <61> Aerobic bacteria: < 5 cfu/200 µL
Yeast/moulds: < 5 cfu/200 µL

1Unit-definition: One unit (U) will digest salmon sperm DNA to acid-soluble oligonucleotides equivalent to a ΔA260nm of 1.0 in 30 min at pH 8.0 at 37 °C.

Please refer to the Validation Guide for more information on the enzyme activity.

Stability & Storage Conditions

When stored under recommended conditions (-20 °C ± 5 °C), the enzyme is stable for at least 12 months. The long-term stability of DENARASE® High Salt is currently under investigation.
Note: It is not recommended to store the product at -70 °C or below, since freezing the product will cause loss of activity.

Packaging Information

DENARASE® High Salt is filled in non-pyrogenic, USP Class VI compliant vials. The product vials are shipped under qualified cooled conditions. The shipping temperature may differ from the recommended storage temperature without affecting product quality. All DENARASE® High Salt products will be delivered by c-LEcta in a sealed secondary packaging with tamper-evident seals.

Get in touch with our DENARASE® expert team!

Vaishnavi Devarakonda

Vaishnavi Devarakonda
DENARASE® Sales

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Melissa Rangel

Melissa Rangel
DENARASE® Sales

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Sedef Ozyurek

Sedef Ozyurek
DENARASE® Sales

Need support? Contact me!