| Expression System |
E.coli
|
| Species |
Serratia Marcescen
|
| Tag |
N-terminal 6 × His Tag
|
| Theoretical Molecular Weight |
27.5 kDa
|
| Biological Activity |
≥ 1.1×106
U/mg;
One unit
of Benz-Neburase™, His is defined as the amount of enzyme that causes a ∆A260
of 1.0 (equivalent to the complete digestion of 37μg DNA) in 30 min. |
| Formulation |
Supplied
as a solution of 20 mM Tris-HCl, 2 mM MgCl2, 20 mM NaCl, 50% Glycerol, pH 8.0.
|
| Apparent Molecular Weight |
~27.5 kDa, on SDS-PAGE
under reducing conditions
|
| Storage & Stability |
This product remains stable for up to 2 weeks at 4 °C or up to 24 months at -20 °C.
Avoid repeated freeze-thaw cycles.
Do not store below -20 °C! |
This
enzyme can be used to remove all forms of DNA and RNA, including double
stranded, single stranded, linearized, and circular forms.
|
The
activity of Benz-Neburase™, His requires 1-2 mM Mg2+.
|
| Purity |
≥ 95% as analyzed by reducing SDS-PAGE
|
| Enzyme Activity |
≥
250 U/μL
|
| Endotoxin Level |
≤ 0.1 EU/kU as determined by gel clotting method.
|
Lane M：DNA marker
Lane 1：GenScript Benz-Neburase™, His + PCR product
Lane 2：PCR product
Lane 3：GenScript Benz-Neburase™, His + Genomic DNA
Lane 4：Genomic DNA
Lane 5：GenScript Benz-Neburase™, His + Plasmid DNA
Lane 6：Plasmid DNA
Lane 7：GenScript Benz-Neburase™, His + RNA
Lane 8：RNA
Lane M：SDS-PAGE marker
Lane 1：Reducing ( R)
Lane 2：Non-reducing (NR)
Purity: > 95% as analyzed by SDS-PAGE
| Target Background |
The Benz-Neburase is a
genetically engineered endonuclease from Serratia
marcescens used as a DNA eraser in the purification processes of biological
molecules. The enzyme cleaves all forms of DNA and RNA into smaller nucleotides
of around 5-8 base pairs. Benz-Neburase requires divalent cation, preferably Mg2+
for activity, displays a broad pH tolerance, ranging from pH 6 to pH10, optimal
at 8-8.5, and has a wide temperature tolerance, ranging from 35℃ to 44℃. The nuclease is a physiologic
homodimer and functions more progressively than the monomer. Two disulfide
bonds in the nuclease are crucial to its activity and stability. The enzyme is
active in a broad range of conditions and is free of proteolytic activity. This
makes the enzyme especially useful for applications with challenging DNA
contents such as lysed host cells in viral vector manufacturing processes.
|
| Synonyms |
alternative to Benzonase®
Benzonase
is a registered trademark of Merck KGaA |
| References |
1. Nestle, Marion, and W. K. Roberts. "An
extracellular nuclease from Serratia marcescens: I. Purification and some
properties of the enzyme." Journal of Biological Chemistry 244.19
(1969): 5213-5218.
2. Benedik, Michael J., and Ulrich Strych.
"Serratia marcescens and its extracellular nuclease." FEMS
microbiology letters 165.1 (1998): 1-13.
3. Filimonova, Maria N., Kurt L. Krause, and
Michael J. Benedik. "Kinetic studies of the Serratia marcescens
extracellular nuclease isoforms." Biochemistry and molecular
biology international 33.6 (1994): 122-1032.
4. Friedhoff, Peter, et al. "A procedure for
renaturation and purification of the extracellular Serratia marcescens nuclease
from genetically engineered Escherichia coli." Protein expression
and purification 5.1 (1994): 37-43.
5. Franke, Ingo, Gregor
Meiss, and Alfred Pingoud. "On the Advantage of Being a Dimer, a Case
Study Using the Dimeric Serratia Nuclease and the Monomeric Nuclease from Anabaena
sp. Strain PCC 7120." Journal of Biological Chemistry 274.2
(1999): 825-832. |
For laboratory research use only. Direct human use,
including taking orally and injection and clinical use are forbidden.
