Benzonase Nucleases

Introduction

Benzonase is a genetically engineered nucleic acid endonuclease from Serratia marcescens. It degrades all forms of DNA and RNA (including single-stranded, double-stranded, linear and cyclic) without protein cleavage activity and with high specificity over a wide range of conditions. It completely digests nucleic acids into 5'-monophosphate oligonucleotides of 3-5 bases in length (below the hybridization limit), making it suitable for the removal of nucleic acids from recombinant proteins and high-density cell fermentation viral fluids, in accordance with FDA protocols for handling nucleic acid contamination. The ability to rapidly hydrolyze nucleic acids makes this enzyme the best choice for reducing viscosity to reduce processing time and increase protein/antigen yield.

Application

• Use with cell or bacterial lysates to remove nucleic acids from crude extracts, reduce solution viscosity, and improve protein/antigen yields.
• When purifying recombinant proteins or extracting proteins from tissue cell samples, B nuclease can effectively reduce sample viscosity and facilitate downstream operations.
• In the clarification of viral solution in cell culture, B nuclease can effectively reduce the nucleic acid in viral culture solution and reduce the clustering of DNA and RNA with virus. At the same time, it can reduce the viscosity of the stock solution, which can improve the speed of stock solution clarification and filtration capacity.
• Removal of nucleic acid contamination during protein extraction

Q&A

1. In which step is it added Benzonase Nucleases ?
   BenzoNuclease can be added directly to the cell/bacterial lysate for action. Note that if the lysate does not contain Mg2+, Mg2+ needs to be added to 1-2 mM.
2. How can I ensure the digestion of BenzoNuclease when the reaction temperature is below 37°C?
   The digestion of Benzonase Nucleases II depends on the amount of enzyme, the reaction temperature and the reaction time. When the reaction temperature is low, it is more recommended to extend the reaction time to ensure the digestion of Benzonase Nucleases II in order to avoid residual problems caused by too much Benzonase Nucleases .
3. What are the inhibition conditions for Benzonase Nucleases?
   Benzonase Nucleases can remain active under a wide range of conditions, but 1-2 mM Mg2+ is critical for its activity.
   In general, Benzonase Nucleases activity can be inhibited by high salts, e.g. > 500 mM monovalent cations (e.g. Na+, K+, etc.), > 100 mM guanidine HCl, > 100 mM phosphate, > 100 mM ammonium sulfate etc. In addition, chelating agents can also inhibit enzymatic activity by chelating Mg2+ in the system, and by adding more Mg2+ the activity of Benzonase Nucleases can be restored (1 mM EDTA partially inhibits Benzonase Nucleases II activity and 5 mM EDTA causes about 90% loss of activity).
4. Is Benzonase Nucleases compatible with protease inhibitors?
   Yes. However, it is important to note that EDTA-free protease inhibitors are preferred (≥1 mM EDTA inhibits Benzonase Nucleases activity).

5. How do I remove Benzonase Nucleases ?
   Benzonase Nucleases is fused with a 6×His tag, which can be removed by a nickel column. In addition, Benzonase Nucleases can also be removed in combination with the purification process of the manufactured product itself, e.g. by TFF (tangential flow filtration), dialysis and chromatography (IEX, SEC, HIC), ultrafiltration, etc.
6. Are there any patent restrictions on the application of Benzonase Nucleases ?
   No. There are no patent restrictions.
7. What is the best method to detect nucleic acid residues after Benzonase Nucleases treatment?
   The qPCR method is recommended. The sensitivity and reproducibility of the qPCR method are good, and it can be quantified to have a more accurate grasp of the nucleic acid residues.
8. The storage solution of GMP-1707 contains 2mM MgCl2, do I need to add magnesium ions after adding to the sample?
   If the sample solution does not contain MgCl2 or the concentration of MgCl2 is low, it is recommended to add MgCl2 to the optimal working concentration (1-2mM) of the allosteric nuclease.