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Manufactured By: Sino Biological Inc.
Synonym: SuperNuclease, Nuclease, Serratia marcescens' extracellular endonuclease, Benzonase®; equivalent (Benzonase®; endonuclease is a trademark of Merck KGaA, Darmstadt, Germany).
Protein Construction: Construction of SuperNuclease is similar to its natural form.
Source: Serratia marcescens
Expression Host: Escherichia coli (E. coli).

Purity: > 98%, as determined by SDS-PAGE
Endotoxin: Less than 0.22 EU/mg of the SuperNuclease (SSNP01) as determined by the LAL method.
Stability: Samples are stable for up to twelve months from date of receipt -70℃.
Predicted N terminal: Three isoforms with different N terminal may be found from the compound—Sm1 (22D-266N), Sm2 (23T-266N) and Sm3 (25E-266N), the activity analysis shows that they were functionally equivalent[6, 15].


Fig. 1.
The SuperNuclease comprises 266 amino acids and has a calculated molecular mass of Sm1 (26708.2 Da), Sm2 (26591.8 Da) and Sm3 (26376.4 Da) [15].
The apparent molecular mass of SuperNuclease is approximately 26.5 KDa.The apparent molecular mass of each lot were determined by SDS-PAGE (12%) under reducing condition, visualized by coomassie blue staining, and analysed by Quantity One.



Fig. 2.
The high purity of SuperNuclease is needed in many applications. Result shows the purity of SuperNuclease was approximately 98 %( SSNP01).
The purity of each lot were determined by SDS-PAGE (12%) under reducing condition, visualized by coomassie blue staining and analysed by BandScan.

Shipment: Shipped at ambient temperature. The liquid or lyophilized enzyme is stable for at least 21d when stored at 37 ℃ (or ambient temperature).
Formulation SuperNuclease is lyophilized from sterile 50 mM Tris-HCl pH 8.0, 20 mM NaCl, 2 mM MgCl2, 5 % trehalose, 5 % mannitol, and 0.01 % Triton® (Triton® is a trademark of Union Carbide, USA). Follow the instructions on the vial. Centrifuge the vial at 4℃ before opening to recover the entire contents. Please contact us for any concerns or special requirements.
Reconstitution: Follow the instructions on the vial. Centrifuge the vial at 4℃ before opening to recover the entire contents. Normaly,25U/μL is recommended to be the final concentration.
Storage: Store it under sterile conditions at -20℃ to -80℃ upon receiving for at least 12 months. Recommend to aliquot the protein into smaller quantities for optimal storage. Avoid repeated freeze-thaw cycles.

(1).Viscosity Reduction of E. coli Lysate

Cell extracts often show high viscosity due to nucleic acids. The viscosity of lysate may lead to reduction of the protein production. This problem can be solved by using SuperNuclease.

Experimental design

E. coli (1.0 g) with a recombined pET-28a construct was suspended in cell lysate buffer (50 mM Tris-HCl (pH 8.0), 4 M Urea, 100 mM DTT, and 1% Triton X-100), and resulted in 1.0 g/mL. Then cell lysate was incubated with SuperNuclease at 4℃ for 5 min. Then samples were centrifuged at 10000 g for 1 min, and photographed.


1cell lysate

Fig. 3.
Viscosity reduction assay shows that the SuperNuclease can reduce the viscosity of E. coil lysate.

A. With 25U/mL SuperNuclease
B. Without SuperNuclease

(2).Nucleic Acid Elimination of Nucleoprotein (NP)

Nucleoprotein of virus is always thought to be an attractive target for vaccine development. Biotech has make it possible to express and purify the NP from the engineered E. coli, Baculovirus-insect cells or other expression hosts.
However, reducing of the nucleic acid contamination is needed for successful development of the vaccine. This problem can be solved by using SuperNuclease, which can decrease the viscosity of the cell lysate, reducing the ratio of A260/A280.

Experimental design

Expression of NP took place over 48 h at 26℃, cell plate was collected by centrifugation, and washed with buffer A (20 mM Tris pH8.0, 50 mM NaCl, 5 mM MgCl2). Cells were lysed by ultrasonic probe in buffer A (25 U/mL SuperNuclease). Supernatant contained NP-(His tag) was collected by centrifugation for 20 min, at 10000 g, 4℃, then NP was adhered to Ni+ column, and washed with low concentration of imidazole, high concentration of NaCl (to separate the fragment of nucleic acid from protein), low concentration of NaCl, and proper concentration of imidazole to elute the protein from the column. Finally, NP was desalted and stored in the proper buffer.


 2NP before