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Literature summary for 3.4.21.1 extracted from

  • Yang, F.; Su, W.; Lu, B.; Wu, T.; Sun, L.; Hara, K.; Cao, M.
    Purification and characterization of chymotrypsins from the hepatopancreas of crucian carp (Carassius auratus) (2009), Food Chem., 116, 860-866.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information during purification, both chymotrypsin A and B are fully activated when extracted Carassius auratus

Application

Application Comment Organism
additional information potential application value of the two chymotrypsins A and B where low processing temperature and higher enzymatic activity is needed Carassius auratus

Inhibitors

Inhibitors Comment Organism Structure
benzamidine partially inhibits both chymotrypsin A and B. At room temperature, at 5 mM concentration, 60.3% and 62.5% residual activity for chymotrypsin A and B, respectively Carassius auratus
Cd2+ partially inactivates both chymotrypsin A and B at concentrations of 1 and 5 mM. At room temperature, at 5 mM concentration, 76.8% and 49.7% residual activity for chymotrypsin A and B, respectively Carassius auratus
chymostatin effectively inhibits both chymotrypsin A and B. At room temperature, at 0.01 mM concentration, no residual activity for both chymotrypsin A and B Carassius auratus
Cu2+ strongly inactivates both chymotrypsin A and B at concentrations of 1 and 5 mM. At room temperature, at 5 mM concentration, 0% and 2.5% residual activity for chymotrypsin A and B, respectively Carassius auratus
EDTA shows minimal inhibitory effect on both chymotrypsin A and B. At room temperature, at 1 mM concentration, 99% and 97% residual activity for chymotrypsin A and B, respectively Carassius auratus
Fe2+ strongly inactivates both chymotrypsin A and B at concentrations of 1 and 5 mM. At room temperature, at 5 mM concentration, 14.6% and 21.1% residual activity for chymotrypsin A and B, respectively Carassius auratus
Mn2+ partially inactivates both chymotrypsin A and B at concentrations of 1 and 5 mM. At room temperature, at 5 mM concentration, 85.3% and 48.9% residual activity for chymotrypsin A and B, respectively Carassius auratus
N-tosyl-L-phenylalanine chloromethyl ketone significant inhibition, especially to chymotrypsin B. At room temperature, at 0.05 mM concentration, 35.6% and 17.5% residual activity for chymotrypsin A and B, respectively Carassius auratus
Pefabloc SC effectively inhibits both chymotrypsin A and B. At room temperature, at 1 mM concentration, 1.2% and 1.1% residual activity for chymotrypsin A and B, respectively Carassius auratus
pepstatin shows minimal inhibitory effect on both chymotrypsin A and B. At room temperature, at 0.03 mM concentration, 95.9% and 92.3% residual activity for chymotrypsin A and B, respectively Carassius auratus
phenylmethanesulfonyl fluoride effectively inhibits both chymotrypsin A and B. At room temperature, at 1 mM concentration, 0.4% and 0.3% residual activity for chymotrypsin A and B, respectively Carassius auratus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0005
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin chymotrypsin B, at pH 7.5 and 37°C Carassius auratus
0.0014
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin chymotrypsin A, at pH 7.5 and 37°C Carassius auratus
0.8
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin at pH 7.5 and 37°C Bos taurus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ both chymotrypsin A and B are slightly activated at concentrations of 1 and 5 mM. At room temperature, at 5 mM concentration, 115% and 103% relative activity for chymotrypsin A and B, respectively Carassius auratus
Mg2+ both chymotrypsin A and B are slightly activated at concentrations of 1 and 5 mM. At room temperature, at 5 mM concentration, 111% and 110% relative activity for chymotrypsin A and B, respectively Carassius auratus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27000
-
x * 28000, chymotrypsin A, SDS-PAGE. X * 27000, chymotrypsin B, SDS-PAGE Carassius auratus
28000
-
x * 28000, chymotrypsin A, SDS-PAGE. X * 27000, chymotrypsin B, SDS-PAGE Carassius auratus

Organism

Organism UniProt Comment Textmining
Bos taurus
-
-
-
Carassius auratus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
chymotrypsin A and B, to homogeneity by ammonium sulfate fractionation and gel filtration Carassius auratus

Source Tissue

Source Tissue Comment Organism Textmining
hepatopancreas
-
Carassius auratus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.62
-
crude extract Carassius auratus
4.1
-
after ammonium sulfate fractionation, 6.6fold purified enzyme Carassius auratus
581.6
-
chymotrypsin B, after gel filtration, 938.1fold purified enzyme Carassius auratus
981.8
-
chymotrypsin A, after gel filtration, 1583fold purified enzyme Carassius auratus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Gelatin + H2O chymotrypsin A is active in degrading gelatin, while chymotrypsin B does not reveal activity against gelatin Carassius auratus ?
-
?
additional information chymotrypsin does not reveal activity against gelatin Bos taurus ?
-
?
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O
-
Bos taurus succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
?
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin + H2O chymotrypsin A shows much lower catalytic efficiency to the substrate than chymotrypsin B Carassius auratus succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
-
?

Subunits

Subunits Comment Organism
? x * 28000, chymotrypsin A, SDS-PAGE. X * 27000, chymotrypsin B, SDS-PAGE Carassius auratus

Synonyms

Synonyms Comment Organism
chymotrypsin
-
Bos taurus
chymotrypsin A anionic chymotrypsin Carassius auratus
chymotrypsin B cationic chymotrypsin Carassius auratus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
-
optimum temperatures of chymotrypsin A and B are 40 and 50°C, respectively, at pH 7.5 using 0.1 M Tris-HCl buffer Carassius auratus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 45 chymotrypsin A at 20°C exhibits more than 60% of its maximum activity, which is much higher than chymotrypsin B (27%). Slight changes in activity for both enzymes at 37°C. Relative activities of the two enzymes decrease sharply at above 45°C Carassius auratus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.9
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin at pH 7.5 and 37°C Bos taurus
2.7
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin chymotrypsin A, at pH 7.5 and 37°C Carassius auratus
3.4
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin chymotrypsin B, at pH 7.5 and 37°C Carassius auratus

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
optimal pH of chymotrypsin A and B are 7.5 and 8.0, respectively, at 37°C Carassius auratus

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 11 chymotrypsin A and B. The two chymotrypsins are unstable at pH below 5.0, about 10% activity for the enzymes at pH 4.0 Carassius auratus

General Information

General Information Comment Organism
physiological function anti-chymotrypsin B weakly cross reacts with chymotrypsin A Carassius auratus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1100
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin at pH 7.5 and 37°C Bos taurus
1900
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin chymotrypsin A, at pH 7.5 and 37°C Carassius auratus
6800
-
succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin chymotrypsin B, at pH 7.5 and 37°C Carassius auratus