DENARASE®

For enzymatic DNA removal in Bioprocessing!

Enzyme Characteristics

DENARASE® is a genetically engineered endonuclease from Serratia marcescens. The protein consists of two subunits with a calculated molecular weight of 27 kDa each. The enzyme catalyzes the hydrolysis of phosphodiester bonds between nucleotides (single-stranded, double-stranded, linear, and circular, sequence-independent) leaving short fragments with a length of 2-5 bases with a 5’-monophosphate end.

Operating conditions
DENARASE is a robust enzyme that is active under varying conditions. Like other enzymes its activity depends on various factors like temperature, pH and concentrations of cofactor and inhibitors.

Temperature/pH
In order to determine the optimal reaction temperature and optimal pH value DENARASE activity was measured under standard conditions at different temperatures and with different buffers at different pH values. The optimal reaction conditions for DENARASE are 37 °C at pH 8.0 – 9.0 (see Figs. 1, 2).

Cofactor
DENARASE uses Mg2+ as a cofactor and a minimum amount is needed for enzyme activity. DENARASE requires 1- 2 mM Mg2+ cations for optimal activity (see Fig. 3). A large excess of MgCl2 reduces activity (see Fig. 4).

Stability and Storage Conditions
DENARASE is stable within the specification range at a storage temperature of -20 °C for a period of at least 24 months from the date of product release. 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 is filled in non-pyrogenic, USP Class VI-compliant vials. The product vials are packed in polystyrene boxes and shipped under refrigerated conditions. Shipping temperature may differ from storage temperature without affecting product quality.

Please refer to the Validation Guide, which is available on request, for more information on the enzyme activity.


Effect of Temperature and pH value

  • Effect of Temperature on DENARASE performance

    Fig. 1: Effect of Temperature

  • The Effect of pH value in different buffer systems

    Fig. 2: Effect of pH value in different buffer systems

The effect of low and high Magnesium Chloride

Product Specification

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

CriteriaMethodSpecification
Appearancevisualclear, transparent solution
Activityphotometric> 250 U/µL
PurityProtein purity determined by SDS-PAGE and silver staining≥ 99 %
Specific ActivityActivity per protein content determined photometrically at 280 nm
with a molar extinction coefficient of 44,600 L x mol-1 x cm-1
> 6 x 105 U/mg
Protease activityProtease detection assayNo protease activity detectable
Endotoxin levelLAL-Test acc. to Ph. Eur. 2.6.14, Method C< 0.25 EU/kU
Total microbial countTAMC/TYMC acc. to Ph. Eur. 2.6.12Aerobic bacteria: < 5 cfu/200 µL

Yeast/moulds: < 5 cfu/200 µL

Unit-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.

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