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Literature summary for 3.4.21.1 extracted from

  • Castillo-Yanez, F.; Pacheco-Aguilar, R.; Lugo-Sanchez, M.; Garcia-Sanchez, G.; Quintero-Reyes, I.
    Biochemical characterization of an isoform of chymotrypsin from the viscera of Monterey sardine (Sardinops sagax caerulea), and comparison with bovine chymotrypsin (2009), Food Chem., 112, 634-639.
    View publication on PubMed

Application

Application Comment Organism
biotechnology dependence of activity on pH and temperature makes chymotrypsin II a biotechnological alternative for food processing when low temperatures are needed like in fish ripening, fish sauce production, fish protein hydrolysate production and other emerging processes. This enzyme together with other proteases from sardine viscera may aid in the enzymatic treatment of stick-water, in which a reduction in viscosity is required for further processing of effluent Sardinops caeruleus

Inhibitors

Inhibitors Comment Organism Structure
benzamidine at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, inhibits chymotrypsin II, whereas chymotrypsin I is not inhibited Sardinops caeruleus
EDTA at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, inhibits chymotrypsin II, whereas chymotrypsin I is not inhibited Sardinops caeruleus
additional information at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, is not inhibited by benzamidine and EDTA Bos taurus
N-toluenesulfonyl-L-phenylalanine chloromethyl-ketone at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, partially inhibits (53% residual activity) Bos taurus
N-toluenesulfonyl-L-phenylalanine chloromethylketone at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, partially inhibits chymotrypsin I (56% residual activity) and chymotrypsin II (60% residual activity) Sardinops caeruleus
phenylmethylsulfonyl fluoride at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, fully inhibits chymotrypsin I and chymotrypsin II (1-2% residual activity) Sardinops caeruleus
phenylmethylsulfonyl-fluoride at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, fully inhibits (4% residual activity) Bos taurus
Soybean trypsin inhibitor at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, fully inhibits (17% residual activity) Bos taurus
Soybean trypsin inhibitor at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, fully inhibits chymotrypsin I and chymotrypsin II (no residual activity) Sardinops caeruleus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.048
-
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 30°C, chymotrypsin II Sardinops caeruleus
0.074
-
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 30°C, chymotrypsin I Sardinops caeruleus
0.09
-
succinyl-L-Ala-Ala-Pro-L-phenylalanine-p-nitroanilide at 30°C Bos taurus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
25500
-
gel filtration Sardinops caeruleus

Organism

Organism UniProt Comment Textmining
Bos taurus
-
-
-
Sardinops caeruleus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
chymotrypsin I and II, by ionic exchange chromatography Sardinops caeruleus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
36
-
at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, on the amide substrate Bos taurus
39
-
chymotrypsin II, on the amide substrate, at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2 Sardinops caeruleus
58
-
chymotrypsin I, on the amide substrate, at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2 Sardinops caeruleus
84
-
at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2, on the ester substrate Bos taurus
109
-
chymotrypsin II, on the ester substrate, at 25°C, in 50 mM Tris-HCl buffer and 10 mM CaCl2 Sardinops caeruleus
168
-
chymotrypsin I, on the ester substrate, at 25°C, pH 8.0, in 50 mM Tris-HCl buffer and 10 mM CaCl2 Sardinops caeruleus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzoyl-L-tyrosine ethyl ester + H2O
-
Bos taurus ?
-
?
benzoyl-L-tyrosine ethyl ester + H2O
-
Sardinops caeruleus benzoyl-L-tyrosine + ethanol
-
?
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O
-
Sardinops caeruleus succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
-
?
succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide + H2O
-
Bos taurus succinyl-L-Ala-L-Ala-L-Pro-L-Phe + p-nitroaniline
-
?

Synonyms

Synonyms Comment Organism
chymotrypsin
-
Bos taurus
chymotrypsin I
-
Sardinops caeruleus
chymotrypsin II
-
Sardinops caeruleus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45 50 activity of chymotrypsin I and II increases with temperature up to an optimum of 50 and 45°C, respectively, when assayed against succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at pH 8.0 Sardinops caeruleus
55
-
activity increases with temperature up to an optimum of 55°C, when assayed against succinyl-L-Ala-Ala-Pro-L-phenylalanine-p-nitroanilide at pH 8 Bos taurus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 50 chymotrypsin I shows high activity in the range from 20°C to 50°C and chymotrypsin II from 10°C to 45°C Sardinops caeruleus
55 70
-
Bos taurus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
chymotrypsin I is completely inactivated at 55°C and chymotrypsin II at 50°C Sardinops caeruleus
40
-
is inactivated at 60°C Bos taurus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.9
-
succinyl-L-Ala-Ala-Pro-L-phenylalanine-p-nitroanilide at 30°C Bos taurus
4.8
-
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 30°C, chymotrypsin II Sardinops caeruleus
18.6
-
succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 30°C, chymotrypsin I Sardinops caeruleus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
at 25°C, with succinyl-L-Ala-Ala-Pro-L-phenylalanine-p-nitroanilide as substrate Bos taurus
8
-
chymotrypsin I and II, at 25°C, with succinyl-Ala-Ala-Pro-Phe-p-nitroanilide as substrate Sardinops caeruleus

pH Range

pH Minimum pH Maximum Comment Organism
4 9 low activity chymotrypsin I and II at acidic pH (4–5), considerably activity at pH from 6 to 9 Bos taurus
4 11 low activity of chymotrypsin I and II at acidic pH (4-5), considerably activity at pH from 6 to 9 whilst chymotrypsin I shows the highest activity toward the alkaline pH (7-11) Sardinops caeruleus

pH Stability

pH Stability pH Stability Maximum Comment Organism
additional information
-
is stable at acid and alkaline pH Bos taurus
6 11 chymotrypsin I and chymotrypsin II, are most stable at alkaline pH Sardinops caeruleus

pI Value

Organism Comment pI Value Maximum pI Value
Sardinops caeruleus chymotrypsin I, electrofocusing
-
4.3
Sardinops caeruleus chymotrypsin II, electrofocusing
-
5.8

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
21
-
succinyl-L-Ala-Ala-Pro-L-phenylalanine-p-nitroanilide at 30°C Bos taurus