| Field | Specification |
|---|---|
| Concentration | |
| Endotoxin Level | |
| Expression System | |
| Form | Liquid solution |
| Formulation | |
| Molecular Weight | |
| Product Type | |
| Shipping | |
| Source | Recombinant, engineered from Serratia marcescens, expressed in Bacillus sp. |
| Species | |
| Storage | |
| UniProt # |
About DENARASE® High Salt
How DENARASE® High Salt works
DENARASE® High Salt is an engineered version of the wild-type Serratia marcescens endonuclease. By introducing a few targeted amino acid substitutions, the enzyme was optimized for salt tolerance, retaining activity at higher salt concentrations without losing specificity for nucleic acids. DENARASE® High Salt exhibits high DNA removal activity across a broad range of salt concentrations (0–500 mM NaCl) and pH levels, providing greater flexibility in bioprocessing and enabling seamless integration into existing workflows.
Production of DENARASE® High Salt
DENARASE® High Salt is produced using a method nearly identical to the established, patented DENARASE® manufacturing process, based on expression in a gram-positive Bacillus sp. strain. Developed for commercial production of biologicals, the enzyme is manufactured under GMP conditions complying with EU GMP regulations. Produced without antibiotics, Triton X-100, animal-derived or TSE/BSE risk raw materials.
DENARASE® High Salt is available in two quality grades
DENARASE® High Salt for Research and Development (R&D) use & DENARASE® High Salt for manufacturing under GMP.
- DENARASE® High Salt R&D-grade: produced under ISO 9001 standard; less stringent requirements for documentation, storage, and distribution.
- DENARASE® High Salt GMP-grade: manufactured under EU GMP conditions; dedicated regulatory support via US FDA Drug Master File.
- Both quality grades are technically equivalent, enabling a seamless transition from R&D to GMP manufacturing.
- Product Specification DENARASE® High Salt GMP-grade (PDF)
- Product Specification DENARASE® High Salt R&D-grade (PDF)
- Validation Guide (REQUEST-VALIDATION-GUIDE)
- Comparison Data: DENARASE® High Salt vs. Other Salt-Active Endonucleases (REQUEST-REQUEST-DOWNLOAD-COMPARISON-DENARASE-HIGHSALT)
- US FDA DMF Support Document (REQUEST-DMF-SUPPORT)
DENARASE® High Salt — Enzyme Characteristics
DENARASE® High Salt is engineered to maintain high DNA removal activity across a broad range of salt concentrations (0–500 mM NaCl) and pH levels. Magnesium (Mg2+) is an essential cofactor: optimal DENARASE® High Salt activity is observed between 5–25 mM Mg2+. The enzyme is highly effective at process-relevant pH 7.4 with standard magnesium supplementation (5 mM MgCl2).
Salt & Magnesium Concentration
Fig. 1: Effect of NaCl, pH and magnesium
Fig. 2: Effect of magnesium
pH & Temperature
Fig. 3: Effect of pH value in different buffer systems
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 |
| Temperature Optimum | 37 °C |
| Isoelectric Point (pI, calculated) | 7.83 |
| Active NaCl Range | 0–500 mM |
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 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 × mol−1 × cm−1 | > 4 × 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 |
Storage & Conditions
When stored under recommended conditions (−20 °C ± 5 °C), the enzyme is stable for at least 12 months. Long-term stability is currently under investigation. Note: It is not recommended to store the product at −70 °C or below, as deep freezing will cause loss of activity.
Packaging Information
DENARASE® High Salt is filled in non-pyrogenic, USP Class VI compliant vials. 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 are delivered by c-LEcta in a sealed secondary packaging with tamper-evident seals.
DENARASE® High Salt is an engineered variant of the standard enzyme. A few amino acid substitutions increase salt tolerance without affecting nucleic acid specificity.
No. Both products address distinct application needs and will continue to co-exist in the portfolio.
c-LEcta published comparative data covering the development of DENARASE® High Salt and its performance vs. other commercially available salt-tolerant endonucleases for viral vector manufacturing. Available upon request via the sales team.
The estimated delivery time is 2–3 weeks worldwide.
Hydrolyzes phosphodiester bonds leaving ~3–5 bp fragments. Active on all nucleic acid forms (ss/ds, linear, circular, supercoiled). Engineered salt tolerance makes it ideal for bioprocess steps at elevated ionic strengths.
Recommended for processes above 150 mM NaCl.
Mg2+ must be raised to ≥5 mM (range 5–25 mM depending on NaCl and pH; 15 mM suggested for initial tests). Starting concentration: 10–100 U/mL.
Above 150 mM NaCl as a general rule. For 150–250 mM, test both enzymes — pH and Mg2+ also influence activity in this range.
Viral Vector Production (AAV, Lentivirus / HEK293): used during/after cell lysis in high-salt buffer conditions to reduce viscosity and facilitate purification.
Vaccine Manufacturing (live attenuated, inactivated, VLP): reduces host cell DNA under elevated ionic strength, enhances process robustness.
Two grades — both technically equal with the same specification parameters.
R&D grade: ISO 9001; less strict documentation and distribution requirements.
GMP grade: EU GMP; US FDA Drug Master File support; GDP-compliant distribution.
Yes. The DENARASE® ELISA Kit can quantify DENARASE® High Salt, but since the standard is calibrated for the wild-type enzyme, a correction factor of 1.46 must be applied. Alternatively, generate a separate High Salt standard curve.
Anion exchange, cation exchange, hydrophobic interaction, hydroxyapatite, or size exclusion chromatography. Filtration techniques can also apply. Media selection must be evaluated per specific case.
The DENARASE® ELISA Kit detects residual enzyme to 12 pg/mL via monoclonal antibodies.
Potassium phosphate (200–300 mM) can also quench activity. EDTA (>5 mM) removes free Mg2+ ions and inhibits the reaction.
Yes. All batches must meet the endotoxin specification (<0.25 EU/kU) prior to product release. See the Validation Guide for full details.
A selection of peer-reviewed publications and reports referencing or featuring DENARASE®.
Viral Vectors for Gene Therapy and Vaccine Production
22 publicationsProduction of Virus-like Particles (VLP)
10 publicationsProduction of Bacteriophages
2 publicationsBiofilm Removal
2 publicationsProtein Purification
5 publicationsResearch budgets are tight — we get it. That's why we've put together a fresh round of exclusive promotions designed to help you stock up on the reagents, kits, and consumables your lab depends on, without stretching your budget.
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