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Native endopeptidase Lys-C from Achromobacter lyticus. Lysyl endopeptidase is a serine protease secreted by soil bacteria and specifically hydrolyzes amide and peptide ester bonds at the carboxylic side of lysine and S-aminoethylcysteine residues.
Catalogue Number
324796
Brand Family
Calbiochem®
References
References
Jekel, P.A., et al. 1983. Anal. Biochem. 134, 347.
Product Information
CAS number
78642-25-8
Unit of Definition
One AU (amidase unit) is defined as the amount of enzyme that catalyzes the release of 1 µM <i>p</i>NA from <i>N</i>-benzoyl-DL-lysine-<i>p</i>NA per min at 30°C, pH 9.5.
Jekel, P.A., et al. 1983. Anal. Biochem. 134, 347.
数据表
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
19-June-2008 RFH
Description
Native endopeptidase Lys-C from Achromobacter lyticus. Lysyl endopeptidase is a serine protease secreted by soil bacteria and specifically hydrolyzes amide and peptide ester bonds at the carboxylic side of lysine and S-aminoethylcysteine residues. Optimal pH: 9.0-9.5.
Form
Lyophilized
Recommended reaction conditions
Amidase Activity:Materials Required:
•Reagent A: 200 mM AMP, pH 9.5 (dissolve 4.2 g 2-amino-2-methyl-1,3-propanediol in 150 ml H2O; adjust the pH to 9.5 with 1 M HCl; add H2O to a final volume of 200 ml)
•Reagent B: 2.5 mM Bz-Lys-pNA (dissolve 22.6 mg Nα-benzoyl-DL-lysine-p-nitroanilide in 900 ml H2O)
•Reagent C: enzyme diluent (dissolve 240 µg 2-amino-2-hydroxymethyl-1,3-propanediol in 900 ml H2O; adjust the pH to 8.0 with 0.1 M HCl; add H2O to a final volume of 1 L
•Reagent D: enzyme solution (dissolve 1 vial enzyme in 1 ml H2O; dilute 100 µl to a final volume of 25 ml with Reagent C)
•45% Acetic Acid (stop solution)
Protocol
1. Add 2.6 ml Reagent A to 300 µl Reagent B and pre-incubate at 30°C to equilibrate the temperature.
2. Add 100 µl Reagent D and incubate for 25 min at 30°C. Include 100 µl Reagent C alone as a control.
3. Add 1 ml 45% acetic acid to stop the reaction.
4. Measure the absorbance at 405 nm.
5. Calculate the activity:
Amidase Activity (AU/vial) = [(Atest-Acontrol)/25] x [1/9.62] x [4/0.1] x 250
CAS number
78642-25-8
Specific activity
≥2 AU/mg protein
Unit definition
One AU (amidase unit) is defined as the amount of enzyme that catalyzes the release of 1 µM pNA from N-benzoyl-DL-lysine-pNA per min at 30°C, pH 9.5.
Solubility
H₂O or Tris-HCl buffer, pH 4-11. Reconstitute 1 vial with 1 ml solvent; if the enzyme is not completely solubilized, add 1-3 ml additional solvent until completely dissolved.
Storage
Avoid freeze/thaw
-20°C
Do Not Freeze
Ok to freeze
Special Instructions
Following reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 1 year at -20°C.
Toxicity
Standard Handling
References
Jekel, P.A., et al. 1983. Anal. Biochem. 134, 347.
