Product Specification
| Synonyms | Kexin; Protease KEX2 |
| Expression System | Pichia pastoris |
| Molecular Weight | 75 kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Tag | No Tag |
| Physical Appearance | Lyophilized Powder |
| Storage Buffer | 20mM NaAC-HAC，2mM Ca2+，ph5.2 @ 25°C |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | 1. Kasai N .Identification of the fourth member of the mammalian endoprotease family homologous to the yeast Kex2 protease[J].Journal of Biological Chemistry, 1992, 267(1):45-63.
2. Matsuo T H . Characterization of KEX2-encoded endopeptidase from yeast Saccharomyces, cerevisiae[J].Biochemical and Biophysical Research Communications, 1989. |
Background
Kex2 protease, a precursor processing protease derived from yeast, is a calcium-dependent serine protease that specifically recognizes and cleaves the carboxy-terminal peptide bond of the bi-alkaline amino acids Arg-Arg, Lys-Arg, and Pro-Arg. Unlike trypsin, Kex2 does not recognize and cleave the carboxy-terminal peptide bond of a single basic amino acid, arginine or lysine. In yeast, Kex2 protease is responsible for the processing of killer toxin and α-factor precursors. Kex2 protease activity is not inhibited by conventional serine protease inhibitors such as peptidyl peptidase, PMSF and TPCK. The recombinant Kex2 protease is produced by expression of Saccharomyces cerevisiae and has the same enzyme specificity as the natural Saccharomyces cerevisiae Kex2 enzyme. The optimal pH for action is pH 9.0, and the pH for stable storage is pH 5.0-6.0.
Components
Kex2 lyophilized
Dissolved buffer: 20mmol/L NaAc-HAc, 2mmol/L Ca2+, pH 5.0~5.5
Reaction buffer:50 mM Tris-HCl、2mM Ca2+、(pH 8.0 @ 25°C)
Protocol
Kex2 should be reconstituted by the addition of 20–100 μl of Dissolved buffer. Typical reaction conditions are as follows:
Combine 500 ug of sample with reaction buffer *（Recommended 50 mM Tris-HCl、2mM Ca2+、(pH 8.0 @ 25°C))）
Add 5μL(5μg)of Kex2，Incubate at 25°C for 3 hours
Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate Recommended ratio: protease: fusion protein amount ratio 1:20~1:1000
Guidelines
1.Reaction buffer: pH 7.0-9.0, 50mM Tris-HCl, 2mM Ca2+;
2.If the lyophilized powder cannot be used immediately after dissolution, it is recommended to dissolve the lyophilized powder with 20mM (pH5.2) NaAc-HAc, 2mM Ca2+ buffer;
3.The final concentration of the enzyme after dissolution will be stored at -20℃ for storage after dispensing as needed;
4.The optimal reaction pH for Kex2 is pH 9.0, and the optimal pH for storage is 5.0-6.0.
Unit Definition
One unit refers to the amount of enzyme required to catalyze the conversion of 1 µmol of BOC-Gln-Arg-Arg-Arg-PNA to the product per minute at 25°C, pH 8.0.
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