DENARASE® High Salt - Enzyme Characteristics

Cofactor
Magnesium (Mg2+) is an essential cofactor and prerequisite for the activity of DENARASE® High Salt. Requiring the presence of minimal magnesium levels for basal activity, DENARASE® High Salt shows optimal activity at
5-25 mM Mg2+. However, excess of MgCl2 will reduce DENARASE® High Salt activity (see Fig. 2).

Stability and Storage Conditions
When stored properly (-20 °C ± 5 °C), the enzyme is stable for at least 6 months from the date of quality control release. The long-term stability of DENARASE® High Salt is currently under investigation. 

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.

DENARASE® expert team

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

Salt & Magnesium Concentration

DENARASE® High Salt has been designed to enable DNA clearance activity at higher, process relevant, salt conditions (see Fig. 1). The engineered enzyme exhibits activity from 0-500 mM monovalent cation concentrations.

Effect of Naci

Fig. 1: Effect of NaCl

Effect of Magnesium

Fig. 2: Effect of Magnesium

pH & Temperature

Effect of ph

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

Effect of Temperature-1

Fig. 4: Effect of Temperature

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
≥ 99%
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
>5 x 105 U/mg
Endotoxin Level LAL-Test acc. to Ph. Eur. 2.6.14, Method C <0.25 EU/kU
Total Microbial Count TAMC/TYMC acc. to Ph. Eur. 2.6.12 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.

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Vaishnavi Devarakonda

Vaishnavi Devarakonda
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Melissa Rangel
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