Staphylococcus aureus Proteinase V8 CAS NO.66676-43-5

PRODUCT ADVANTAGE
Sequence grade recombinant protein: high specificity,good stability.
Animal origin free: Recombinant V8 Protease is no exogenous virus contamination,and any animal origin material is not used in the production process.
Stable quality: Mass production can ensure stable and continuous batch production.It is no difference between the batch and the product quality is stable.
High purity: Higher specific activity.Host protein residues is less than the limits of biological products.
Lyophilized powder: The product is lyophilized powder and is easy to store and transport.
Compliance with regulatory requirements: Production equipment and production environment comply with relevant regulatory requirements, and the production process is in full compliance with NSF ISO 9001: 2015 quality system and GMP guidelines.
Complete quality documents: we can provide relevant regular support files in according to customers’ requirement.

 

 Sequencing grade V8 Protease

Catalog Number:V813

CAS:66676-43-5

EC:3.4.21.19

Full Name: Sequencing grade recombinant  V8 protease; Staphylococcus aureus strain V8

Synonyms: V8 protease; Endoproteinase Glu-C; Endoproteinase Asp-C 

Storage temperature: 2-8°C

Source: Recombinant V8 protease is a genetically engineered protein expressed in E.coli

 

1. DESCRIPTION   

Recombinant V8 protease is the serine protease family and can specifically hydrolyzing glutamic acid or aspartic acid residues carboxy side peptide bonds. The Glu carboxy terminal peptide bond can be identified and cleaved in CH₄COONH₄ buffer (pH 7.8) and NH₄HCO₃ (pH 4.0 ). Identify and cleave the Glu or Asp carboxy terminal peptide bond in phosphate buffer (pH 7.8), and the hydrolysis rate of Glu was higher than that of Glu Asp. Recombinant V8 protease has activity between pH 4.0-10.0, and the optimum pH is 8.0-8.5. It can be used alone or in combination with other proteases when used in protein digestion, peptide profiling and peptide mass fingerprinting analysis. The inhibitors of V8 protease are diisopropylfluorophosphate (DFP), α2-macroglobulin and Nα-P-tosyl-L-lysine chloromethyl ketone (TLCK).

 

2. MAIN FEATURES

Source	Recombinant E. coli
Appearance	White or off white, or yellowish powder
Specific activity	≥5.0 U /mg pro.
Purity(SDS-PAGE)	Single major band
Molecular Weight(SDS-PAGE)	24.0±2.4 kDa

UNIT DEFINITION: One unit is defined as the amount of enzyme required to catalytic substrate Z-Phe-Leu-Glu-4- nitranilide produced 1 μmol of 4-nitroaniline per minute at 25°C and pH 7.8 .

SPECIFIC ACTIVITY: At 25°C, the activity measured with Z-Phe-Leu-Glu-4-nitranilide as a substrate is about 20U/mg pro. (Corresponding to the activity measured with casein is about 500 U / mg pro.at 37°C).

 

3. STABILITY OF STORAGE AND TRANSPORT

Stability of storage: Recombinant V8 protease should be stored under 2-8°C in sealed container.It is stable within 6 months after dissolved with1 mM HCl and 50mM HAC at -20°C or below. It is stable within 2 months after dissolved with 1 mM HCl and 50 mM HAC at 4°C.85% activity above can be kept after dissolved with 1 mM HCl and 50 mM HAC at 25°C or 37°C 12 h. It is no activity loss after 10 times repeated freezing and thawing.

Stability of transport: The product is stable by blue ice insulation transport.

 

4. RECOMMEND USAGE

Note that before using

1) Divide the material in according to usage quantity to avoid pollution or self-cutting.

2) Dissolve target protein.

The target protein is dissolved by cleavage buffer, for example 25-50 mM NH₄HCO₃ pH 7.8, if the solubility of target protein is not good, the target protein will be denatured by adding urea, SDS, DTT or heating. The effects of urea and SDS on V8 protease are shown in the

 

Table 1.

3) digest

The recommended ratio of V8 protease and target protein is 1:20-1:100 (W/W).At 25 °C or 37 °C digest the protein 2-18h.

Table 1:Stability of V8 protease in the different concentrations of denaturant, urea 25°C 2 h, SDS 25°C12 h

denaturant	concentration	rest of activity
No any denaturant	0	100%
urea	2M	95%
	5M	12%
	8M	0
SDS	0.1%	90%
	0.5%	80%
	1%	50%

 

4) Application example: recombinant V8 protease digests insulin

100 μl of insulin solution +198 μl 0.4 M Tris-HCl pH 8.0 + 60μl 0.1% V8 Protein solution + ultrapure water to 400 μl. 37 °C water bath reaction 2 h, plus 6μl phosphoric acid solution to terminate the reaction, C18 column analysis:

                A:Before digesting                                                         B:After digesting

 

5.PRODUCT ADVANTAGE

1）Sequence grade recombinant protein: high specificity,good stability.

2）Animal origin free: Recombinant V8 Protease is no exogenous virus contamination,and any animal origin material is not used in the production process.

3）Stable quality: Mass production can ensure stable and continuous batch production.It is no difference between the batch and the product quality is stable.

4）High purity: Higher specific activity.Host protein residues is less than the limits of biological products.

5）Lyophilized powder: The product is lyophilized powder and is easy to store and transport.

6）Compliance with regulatory requirements: Production equipment and production environment comply with relevant regulatory

      requirements, and the production process is in full compliance with NSF ISO 9001: 2015 quality system and GMP guidelines.

7）Complete quality documents: we can provide relevant regular support files in according to customers requirement.