Benzonase Nuclease, 99%
Escherichia Coli.
Greater than 99.0% as determined by SDS-PAGE.
Description
Benzonase Nuclease Serratia Marcescens Recombinant produced in E.coli is a single, non-glycosylated polypeptide chain containing 245 amino acids and having a molecular mass of 30kDa with 2 essential disulfide bonds.
Benzonase Nuclease is purified by proprietary chromatographic techniques.
Product Specs
The enzyme Benzonase Nuclease, derived from Serratia marcescens, exhibits exceptional activity in breaking down nucleic acids in its surrounding environment. This enzyme is highly effective in eliminating nucleic acid contamination from purified protein samples. It finds applications in various processes such as downstream processing and viscosity reduction. Notably, the release of this nuclease by S. marcescens leads to the destruction of DNA, acting as a mechanism for the self-destruction of the microorganism.
Recombinant Benzonase Nuclease from Serratia marcescens, produced in E.coli, is a single polypeptide chain that lacks glycosylation. It comprises 245 amino acids, weighs 30kDa, and is stabilized by two disulfide bonds. The purification process involves proprietary chromatographic techniques.
Benzonase Nuclease solution consists of 50% glycerol, 50 mM Tris-HCl at a pH of 8.0, 20 mM NaCl, and 2 mM MgCl2.
The purity level is determined to be greater than 99% using SDS-PAGE analysis.
One unit (U) of Benzonase Nuclease is defined as the enzyme quantity required to produce a change in absorbance at 260 nm (ΔA260) of 1.0 in 30 minutes. This corresponds to the complete digestion of 37 micrograms of DNA under standard reaction conditions: 1 mg/ml sonicated DNA substrate in 50 mM Tris-HCl (pH 8.0), 0.1 mg/ml BSA, 1 mM MgCl2, incubated at 37°C.
Escherichia Coli.
Unspecific (DNA, RNA) attacks all nucleic acids (single strand, double strand, circular, supercoiled) with no apparent sequence preference. Final reaction product:
5’-mono-phosphate terminated oligonucleotides (3-5 bases).
Protease Activity: Not detectable
Product Science Overview
Benzonase Nuclease is produced through recombinant DNA technology, where the gene encoding the enzyme is inserted into Escherichia coli (E. coli) for expression. The enzyme is a dimeric protein, meaning it consists of two subunits, each with a molecular weight of approximately 30 kDa . It contains two essential disulfide bonds that are crucial for its stability and activity .
The primary function of Benzonase Nuclease is to hydrolyze nucleic acids into smaller oligonucleotides. It achieves this by cleaving the phosphodiester bonds within the nucleic acid backbone, resulting in 5′-monophosphate terminated oligonucleotides that are typically 2 to 5 bases in length . This degradation process is efficient and rapid, making Benzonase Nuclease an ideal tool for removing nucleic acids from protein samples and reducing viscosity in various biological preparations .
Benzonase Nuclease is utilized in a wide range of applications, including:
- Protein Purification: It is used to remove nucleic acids from recombinant protein preparations, ensuring higher purity and quality of the final product .
- Cell Lysis: The enzyme is often included in lysis buffers to digest nucleic acids released during cell disruption, facilitating the complete release of nuclear proteins .
- Immunoprecipitation: Benzonase Nuclease is employed to release protein complexes from the nucleoplasm and chromatin during immunoprecipitation procedures .
- Decellularization: It is used to remove residual nucleic acids from tissues during decellularization processes, which are essential for tissue engineering and regenerative medicine .
Benzonase Nuclease is active over a broad range of conditions. It functions optimally between pH 6 and 10 and temperatures from 0°C to 42°C. The enzyme requires 1-2 mM magnesium ions (Mg²⁺) for activation and remains active in the presence of various detergents, reducing agents, and other additives . However, its activity is inhibited by high concentrations of monovalent cations, phosphate, ammonium sulfate, or guanidine hydrochloride .
