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P6911 Sigma

Protease from Streptomyces griseus

BioReagent, DNase, RNase, and nickase, none detected (No RNase.)

Synonym: Actinase E, Pronase E

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Properties

Related Categories Application Index, Biochemicals and Reagents, Enzymes, Inhibitors, and Substrates, Modifying Enzymes, Molecular Biology,
grade   for molecular biology
product line   BioReagent
form   powder
mol wt   monomer mol wt ~20 kDa
concentration   ≥4 unit/mg
solubility   water: soluble5 - 20 mg/mL
foreign activity   DNase, RNase, and nickase, none detected (No RNase.)
storage temp.   −20°C

Description

Analysis Note

The protease is incubated for 10 minutes at pH 7.5 at 37°C in a 6 ml reaction volume containing 0.54% casein and 0.041 M potassium phosphate buffer. The reaction is stopped by the addition of 5.0 ml of 0.11 M trichloroacetic acid.

General description

A mixture of at least three proteolytic activities including an extracellular serine protease. In general, serine proteases display a wide range of substrate specificities, which are believed to be mediated by an active site composed of one Asp, one His, and a Ser residue in the molecule. This enzyme prefers to hydrolyze peptide bonds on the carboxyl side of glutamic or aspartic acid.

Other Notes

This protease is completely inactivated by heating above 80°C for 15-20 minutes. This enzyme is more active at a higher pH range, showing the proteolytic activity even in 0.2N NaOH solution.

Packaging

1, 5 g in glass bottle

100 mg in glass bottle

Preparation Note

Collected from culture broth of S. griseus.

Specificity

A mixture of at least three proteolytic activities including an extracellular serine protease. In general, serine proteases display a wide range of substrate specificities, which are believed to be mediated by an active site composed of one Asp, one His, and a Ser residue in the molecule. This enzyme prefers to hydrolyze peptide bonds on the carboxyl side of glutamic or aspartic acid.

Unit Definition

One unit will hydrolyze casein to produce color equivalent to 1.0 μmole (181 μg) of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).

Application

Protease is typically used in nucleic acid isolation procedures in incubations of 0.5-3.0 hours supplemented with 0.2% sodium dodecyl sulfate and 10 mM EDTA.

Suitable for:
• Nucleic acid isolation procedures in incubations
• Degrade protein during nucleic acid purification
• Proteolysis of insoluble protein
• Structural protein studies

This enzyme is more active at a higher pH range than the known alkaline protease, showing the proteolytic activity even in 0.2N NaOH solution. This enzyme is useful for proteolysis of insoluble protein and for structure investigation of protein.

Physical properties

Completely inactivated by heating above 80 °C for 15-20 minutes.

Price and Availability


Biomedical Applications
Protocols & Articles
Peer-Reviewed Papers
15

References

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