|
E.C. 3.4.21.1
On this page:
Physical Properties
Molecular Weight: 25 kDa1 (Bovine)
pI: 8.752 (Bovine)
Extinction coefficient: E1%= 20.4 (280nm)
Chymotrypsin is produced in the acinar cells of the pancreas as the inactive precursor, chymotrypsinogen. α-Chymotrypsin is the predominant form of active enzyme produced from it's zymogen, Chymotrypsinogen A.
In vivo, the rate of hydrolysis of the zymogen by trypsin and by autolysis produces varying amounts of α, π, δ and γ variants.3 α-Chymotrypsin is a serine protease of the peptidase S1 family consisting of 241 amino acid residues. The molecule has three peptide chains: an A chain of 13 residues, a B chain of 131 residues, and a C chain of 97 residues.4
Back to top
Specificity and Kinetics
α-Chymotrypsin from bovine pancreas selectively catalyzes the hydrolysis of peptide bonds on the C-terminal side of tyrosine, phenylalanine, tryptophan, and leucine. A secondary hydrolysis will also occur on the C-terminal side of methionine, isoleucine, serine, threonine, valine, histidine, glycine, and alanine.1
pH optimum: 7.88
(pH 6.0: about 35% of maximal activity, pH 9.3: 40% of maximal activity)8
Temperature Optimum: 50 °C9
Back to top
| Inhibitors |
| Product # |
Product Name |
Add to Cart |
| A8849 |
α2-Antiplasmin from human plasma lyophilized powder, ≥5 inhibitor U/mg protein |
|
| A1153 |
Aprotinin from bovine lung lyophilized powder, 3-8 TIU/mg solid |
|
| A2504 |
6-Aminocaproic acid ≥99% (titration), powder |
|
| A6150 |
α1-Antitrypsin from human plasma salt-free, lyophilized powder |
|
| A8456 |
4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride ~97% |
|
| A9024 |
α1-Antitrypsin from human plasma salt-free, lyophilized powder |
|
| A9285 |
α1-Antichymotrypsin from human plasma ~95% (SDS-PAGE), lyophilized powder |
|
| B1552 |
Bromoenol lactone ≥98% |
|
| C7268 |
Chymostatin microbial |
|
| D0879 |
Diisopropyl fluorophosphate |
|
| E7888 |
N-Acetyl-eglin C recombinant, expressed in proprietary host |
|
| G2417 |
Gabexate mesylate solid |
|
| L2023 |
Leupeptin trifluoroacetate salt ≥90% (HPLC), microbial |
|
| T0256 |
Trypsin inhibitor from bovine pancreas Type I-P, essentially salt-free, lyophilized powder |
|
| T1886 |
Trypsin inhibitor from chicken egg white Type IV-O, Purified Ovoinhibitor |
|
| T4376 |
N-p-Tosyl-L-phenylalanine chloromethyl ketone ≥97% (TLC), powder |
|
| T4385 |
Trypsin inhibitor from turkey egg white Type II-T |
|
| T9253 |
Trypsin inhibitor from chicken egg white Type II-O, Partially purified ovomucoid, containing ovoinhibitor |
|
| T9378 |
Trypsin inhibitor from Phaseolus limensis (lima bean) Type II-L, crude powder |
|
| T9777 |
Trypsin-chymotrypsin inhibitor from Glycine max (soybean) lyophilized powder |
|
| Back to top |
α-Chymotrypsin is also completely inhibited by 10 mM Cu2+ and Hg2+.1
| Substrates |
| Product # |
Product Name |
Add to Cart |
| S7388 |
N-Succinyl-Ala-Ala-Pro-Phe p-nitroanilide |
|
| S9761 |
N-Succinyl-Ala-Ala-Pro-Phe-7-amido-4-methylcoumarin |
|
| 1530 |
N-Acetyl-L-tyrosine ethyl ester monohydrate BioChemika, ≥99.0% (sum of enantiomers, HPLC) |
|
| A3401 |
Ala-Ala-Phe-7-amido-4-methylcoumarin |
|
| A9273 |
Ala-Ala-Val-Ala p-nitroanilide |
|
| 13110 |
Magnesium according to Grignard, ≥99.5%, turnings |
|
| B6125 |
N-Benzoyl-L-tyrosine ethyl ester |
|
| B6760 |
N-Benzoyl-L-tyrosine p-nitroanilide |
|
| 13135 |
N-Benzoyl-L-tyrosine p-amidobenzoic acid sodium salt BioChemika, lyophilized, ≥98.0% (sum of enantiomers, HPLC) |
|
| G3769 |
Glutaryl-Ala-Ala-Phe-4-methoxy-β-naphthylamide |
|
| 49737 |
Glutaryl-L-phenylalanine 7-amido-4-methylcoumarin BioChemika, for fluorescence, ≥98.0% (HPLC) |
|
| 49738 |
N-Glutaryl-L-phenylalanine p-nitroanilide BioChemika, ≥98.0% (HPLC) |
|
| 49742 |
N-(N-Glutaryl-L-phenylalanyl)-2-aminoacridone BioChemika, for fluorescence, ≥98.0% (HPLC) |
|
| M4507 |
4-Methylumbelliferyl p-trimethylammoniocinnamate chloride powder |
|
| 85977 |
N-Succinyl-Ala-Ala-Pro-Phe p-nitroanilide BioChemika, ≥99.0% (HPLC) |
|
| S1899 |
N-Succinyl-Gly-Gly-Phe-p-nitroanilide |
|
| S2628 |
N-Succinyl-L-phenylalanine-p-nitroanilide |
|
| 85992 |
N-(N-Succinyl-L-phenylalanyl)-2-aminoacridone BioChemika, for fluorescence, ≥98.0% (HPCE) |
|
| Back to top |
Solubility and Solution Stability
Chymotrypsin is typically soluble in 1 mM HCl (2 mg/ml), yielding a clear solution.
Reconstitute in 1 mM HCl containing 2 mM CaCl2, aliquot, and store at -20 °C. Autolysis will occur when stored at a higher pH. The presence of calcium is also a stabilizer.5 Frozen aliquots are stable for approximately 1 week. Chymotrypsin is both activated and stabilized by Ca2+. The enzyme is active in the presence of 0.1% SDS and 2 M guanidine hydrochloride.
Back to top
Applications
For peptide digestion, use a ratio (w/w) of approximately 1:60 for chymotrypsin:peptide. Perform peptide digests in 100 mM Tris HCl containing 10 mM CaCl2, pH 7.8, at 30 °C. Self digestion may occur if temperatures above 37 °C are used. A known peptide such as melittin should be used as a control for all experiments. Incubate up to 24 hours at 37-40 °C. Digestion can be terminated by adjusting the pH to 2.0.6,7
| Products |
| Product # |
Product Name |
Add to Cart |
| C4129 |
α-Chymotrypsin from bovine pancreas Type II, essentially salt-free, lyophilized powder, ≥40 units/mg protein |
|
| C7762 |
α-Chymotrypsin from bovine pancreas Type I-S, essentially salt-free, lyophilized powder |
|
| C6423 |
α-Chymotrypsin from bovine pancreas suitable for protein sequencing, salt-free, lyophilized powder |
|
| C3142 |
α-Chymotrypsin TLCK Treated from bovine pancreas Type VII, essentially salt-free, lyophilized powder, ≥40 units/mg protein |
|
| C9134 |
α-Chymotrypsin−Agarose from bovine pancreas lyophilized powder, 2,000-3,500 units/g agarose (One ml gel will yield 65-120 units) |
|
| C5088 |
α-Chymotrypsin−polyethylene glycol ~20 BTEE units/mg protein |
|
| C4879 |
α-Chymotrypsinogen A from bovine pancreas Type II, essentially salt-free, lyophilized powder, ≥40 units/mg solid (after activation to α-chymotrypsin.) |
|
| C1012 |
α-Chymotrypsin bovine Type VI (Inactivated with DFP), lyophilized powder, <0.2 units/mg protein |
|
| Back to top |
Applications
- Enzymes of Molecular Biology, vol. 16, Burrell, M. M., ed., Humana Press (Totowa, NJ: 1993), pp. 277-281.
- Ui, N., Isoelectric points and conformation of proteins. II. Isoelectric focusing of alpha-chymotrypsin and its inactive derivative. Biochim. Biophys. Acta, 229(3), 582-589 (1971).
- Desnuelle, P.: The Structure of Chymotrypsin,The Enzymes, 3rd Ed. Vol. 3, P. Boyer, H. et al., Ed., Academic Press, NY, 185-191, 1960
- Hess, G. P., The Enzymes, 3rd ed., vol. 3, Boyer, P. D., ed., Academic Press (New York, NY: 1971), pp. 213-248.
- Burrell, M.M., Enzymes of Molecular Biology, Vol. 16, 278-281 (1993).
- Spackman, D.H., et al., J. Biol. Chem., 235, 648-659 (1960)
- Kamp, R. M. (1986) Advanced Methods in Protein Microsequence Analysis (Wittmann-Liebold, B., Salnikow, J., and Erdmann, V., eds) pp 8-20, Springer Verlag, Berlin
-
- Asgeirsson, B.; Bjarnason, J.B.; Comp. Biochem. Physiol. B 99B, 327-335 (1991)
- Fernandez, M.; Villalonga Mde, L.; Fragoso, A.; Cao, R.; Villalonga, R.; Biotechnol. Appl. Biochem. 36, 235-239 (2002)
Back to top
|
