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DENARASE® High Salt is a bioengineered endonuclease derived from the wild-type Serratia marcescens, typically used for DNA removal in bioprocesses. Compared with the wild-type DENARASE®, several amino acids of this enzyme have been substituted to enhance its salt tolerance without losing its specificity for nucleic acids. DENARASE® High Salt maintains higher activity over a broader range of salt concentrations and pH values, providing customers with greater process flexibility while improving cost-effectiveness for viral vector manufacturing processes—especially under conditions requiring high salt concentrations. Due to its high similarity to the wild-type endonuclease, DENARASE® High Salt can be detected using the same endonuclease ELISA kits as the wild-type enzyme, such as the DENARASE® ELISA Kit and the Cygnus Endonuclease Residue Detection Kit (EonucleaseGTP ELISA, Catalog No. F960).
DENARASE® High Salt is produced using c-LEcta’s mature and patented DENARASE® production process. This process employs a Gram-positive spore-forming bacterial strain (Bacillus sp.) for expression, which significantly reduces the risk of endotoxin residues. The enzyme is manufactured in compliance with ISO 9001 standards, with no antibiotics, animal-derived materials, or materials posing TSE/BSE risks used in the production process.
Enzyme Characteristics of DENARASE® High Salt
DENARASE® High Salt is specifically designed to achieve DNA-clearing activity at higher, process-relevant salt concentrations. This engineered enzyme exhibits activity across a monovalent cation concentration range of 0–500 mM (see Figure 1).
Figure 1. Effect of NaCl on Enzyme Activity
Magnesium ions (Mg2?) are essential cofactors for ensuring the enzymatic activity of DENARASE® High Salt. DENARASE® High Salt exhibits basal activity at low magnesium ion concentrations (5–25 mM Mg2?), but its enzymatic activity decreases at high magnesium ion concentrations (>25 mM Mg2?). Therefore, when using DENARASE® High Salt, the magnesium ion concentration should be maintained within an appropriate range (see Figure 2).
Figure 2. Effect of Mg2? on Enzyme Activity
Quality Parameters of DENARASE® High Salt
The DENARASE® High Salt product is in liquid form, with a formulation consisting of 20 mM Tris-HCl (pH 7.4±0.2), 250 mM NaCl, 5 mM MgCl?, and 50% (v/v) glycerol.
|
Project |
Method |
Quality Standard |
|
Product Characteristic |
Direct Observation |
Clear and transparent liquid |
|
Enzyme Activity |
Optical Assay |
>250 U/μL |
|
Purity |
SDS-PAG and Silver Staining |
≥ 99% |
|
Specific Activity |
Optical Assay (the enzyme activity per unit protein, calculated based on the molar extinction coefficient of 44,600 L·mol?1·cm?1 at 280 nm) |
>5 x 105 U/mg |
|
Endotoxin |
LAL-Test,refer Ph. Eur. 2.6.14, method C |
<0.25 EU/kU |
|
Total Colony Count |
TAMC/TYMC refer Ph. Eur. 2.6.12 |
Aerobic Bacteria: < 5 cfu/200 μL
Yeast: < 5 cfu/200 μL |
1. Unit Definition: One unit of activity (U) is defined as the amount of enzyme that digests salmon sperm DNA into acid-soluble oligonucleotides corresponding to a ΔA????? value of 1.0 within 30 minutes at 37 °C and pH 8.0.
DENARASE® High Salt Product Parameters
· Molecular Weight: 27 kDa
· Optimal pH: pH 7.4 - 9.0
· Optimal Reaction Temperature: 37 °C
· Isoelectric Point (pI): 7.831
· Cofactor: Mg2?
DENARASE® High Salt Ordering Information
|
Product Name |
Item |
Specification |
|
DENARASE High Salt 25 kU |
22002-25k |
25 kU |
|
DENARASE High Salt 100 kU |
22002-100k |
100 kU |
|
DENARASE High Salt 500 kU |
22002-500k |
500 kU |
|
DENARASE High Salt 1000 kU |
22002-1000k |
1000 kU |
|
DENARASE High Salt 5000 kU |
22002-5000k |
5000 kU |
Follow the official WeChat account of XMJ (WeChat ID: xmjsci), reply with "Nuclease Trial", and fill out the form to apply for a free trial of c-LEcta DENARASE® High Salt. Trials are limited in quantity and available on a first-come, first-served basis!
For more information, please call the customer service hotline of Beijing XMJ (the China distributor of c-LEcta) at 400-050-4006 or visit the website www.gq44.cn to obtain additional materials.
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