Broad Spectrum Endonuclease

Biochemical / physiological behavior.
Digest natural or thermally denatured DNA and RNA.

Broad-spectrum unrestricted nuclease is a genetically engineered endonuclease from Serratia marcescens (Serratia marcescens). The protein is a dimer of 30kDa subunits with two necessary disulfide bonds. The endonuclease cleaves and degrades all forms of DNA and RNA (single-stranded, double-stranded, linear and cyclic) and is effective under a wide range of operating conditions. The best pH of enzyme activity is 8.0-9.2. It completely digests the nucleic acid into 5-monophosphate-terminal oligonucleotides with a length of 3 to 5 bases. This is an ideal method for removing nucleic acids from recombinant proteins and is also suitable for applications that require complete digestion of nucleic acids. It can also reduce the viscosity of protein extract and prevent cells from caking. The pretreatment of protein samples can improve the resolution of protein samples on 2D gel electrophoresis by completely removing nucleic acids from them.

Unit definition.
Under the condition of 37 °C, pH8.0, 37 μ g of substrate DNA (salmon sperm DNA) can be digested into oligodeoxynucleotides (equivalent to 1. 0 change in 260nm absorption) within 30 minutes.

Physical form.
The solution of 50% glycerol contains 20mM Tris HCl,pH 8.0 minute 2mm MgCl2 and 20mM NaCl.

Application.
Used to remove nucleic acid from protein samples.

Broad-spectrum unrestricted nucleases (totipotent nucleases), or endonucleases from Serratia marcescens, can be used to degrade all forms of DNA and RNA without proteolytic activity. Broad-spectrum endonuclease can also be used in microcalorimetric experiments to prepare proteins.

The enzyme has been used to limit cell aggregation in the preparation of chimeric cell mixtures. It is also used to prepare nuclear extracts by digesting DNA and releasing nuclear proteins closely related to DNA.