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Literature summary for 3.4.21.4 extracted from

  • Marcuschi, M.; Esposito, T.S.; Machado, M.F.; Hirata, I.Y.; Machado, M.F.; Silva, M.V.; Carvalho, L.B.; Oliveira, V.; Bezerra, R.S.
    Purification, characterization and substrate specificity of a trypsin from the Amazonian fish tambaqui (Colossoma macropomum) (2010), Biochem. Biophys. Res. Commun., 396, 667-673.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
benzamidine 99% inhibition at 1 mM Colossoma macropomum
EDTA weak inhibition, 18% inhibition at 10 mM Colossoma macropomum
additional information no inhibition by N-alpha-tosyl-L-Phe-chloromethyl ketone, E-64, and O-fenantroline Colossoma macropomum
N-alpha-tosyl-L-Lys-chloromethyl ketone complete inhibition at 1 mM Colossoma macropomum
PMSF 73% inhibition at 1 mM Colossoma macropomum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0005
-
Abz-FRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0006
-
Abz-KRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.001
-
Abz-RSFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.001
-
Abz-LRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0017
-
Abz-RYFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0017
-
Abz-RRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.00185
-
Abz-RHFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0029
-
Abz-VRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0031
-
Abz-RPFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.00325
-
Abz-RTFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0033
-
Abz-WRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0049
-
Abz-RNFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0056
-
Abz-YRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0068
-
Abz-HRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0069
-
Abz-ERFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0074
-
Abz-RQFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0074
-
Abz-NRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0081
-
Abz-RDFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0089
-
Abz-QRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0129
-
Abz-REFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0159
-
Abz-RGFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0163
-
Abz-GRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
0.0183
-
Abz-RVFK-EDDnp pH 8.0, 25°C Colossoma macropomum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23900
-
x * 23900, SDS-PAGE Colossoma macropomum

Organism

Organism UniProt Comment Textmining
Colossoma macropomum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme from the pyloric caecum of tambaqui by heat treatment, ammonium sulfate fractionation, gel filtration, and 4-aminobenzamidine affinity chromatography Colossoma macropomum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Abz-ERFK-EDDnp + H2O
-
Colossoma macropomum Abz-ER + FK-EDDnp
-
?
Abz-FRFK-EDDnp + H2O
-
Colossoma macropomum Abz-FR + FK-EDDnp
-
?
Abz-GRFK-EDDnp + H2O
-
Colossoma macropomum Abz-GR + FK-EDDnp
-
?
Abz-HRFK-EDDnp + H2O
-
Colossoma macropomum Abz-HR + FK-EDDnp
-
?
Abz-KRFK-EDDnp + H2O
-
Colossoma macropomum Abz-KR + FK-EDDnp
-
?
Abz-LRFK-EDDnp + H2O
-
Colossoma macropomum Abz-LR + FK-EDDnp
-
?
Abz-NRFK-EDDnp + H2O
-
Colossoma macropomum Abz-NR + FK-EDDnp
-
?
Abz-QRFK-EDDnp + H2O
-
Colossoma macropomum Abz-QR + FK-EDDnp
-
?
Abz-RDFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + DFK-EDDnp
-
?
Abz-REFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + EFK-EDDnp
-
?
Abz-RGFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + GFK-EDDnp
-
?
Abz-RHFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + HFK-EDDnp
-
?
Abz-RNFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + NFK-EDDnp
-
?
Abz-RPFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + PFK-EDDnp
-
?
Abz-RQFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + QFK-EDDnp
-
?
Abz-RRFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + RFK-EDDnp
-
?
Abz-RSFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + SFK-EDDnp
-
?
Abz-RTFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + TFK-EDDnp
-
?
Abz-RVFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + VFK-EDDnp
-
?
Abz-RYFK-EDDnp + H2O
-
Colossoma macropomum Abz-R + YFK-EDDnp
-
?
Abz-VRFK-EDDnp + H2O
-
Colossoma macropomum Abz-VR + FK-EDDnp
-
?
Abz-WRFK-EDDnp + H2O
-
Colossoma macropomum Abz-WR + FK-EDDnp
-
?
Abz-YRFK-EDDnp + H2O
-
Colossoma macropomum Abz-YR + FK-EDDnp
-
?
additional information the purified trypsin from Colossoma macropomum shows substrate specificity for arginine at P1, efficiently hydrolizing substrates with leucine and lysine at P2 and serine and arginine at P10 Colossoma macropomum ?
-
?
N-benzoyl-DL-arginine 4-nitroanilide + H2O a synthetic trypsin substrate Colossoma macropomum N-benzoyl-DL-arginine + 4-nitroaniline
-
?
N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
Colossoma macropomum N-benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
?

Subunits

Subunits Comment Organism
? x * 23900, SDS-PAGE Colossoma macropomum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
with substrate N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin Colossoma macropomum
70
-
with substrate N-benzoyl-DL-arginine 4-nitroanilide Colossoma macropomum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
1 h, purified enzyme, 20% activity remaining with substrate N-benzoyl-DL-arginine 4-nitroanilide Colossoma macropomum
60
-
3 h, purified enzyme, 60% activity remaining with substrate N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin Colossoma macropomum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.1
-
Abz-RPFK-EDDnp pH 8.0, 25°C Colossoma macropomum
2.33
-
Abz-RDFK-EDDnp pH 8.0, 25°C Colossoma macropomum
4.59
-
Abz-RVFK-EDDnp pH 8.0, 25°C Colossoma macropomum
6.32
-
Abz-WRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
7.49
-
Abz-REFK-EDDnp pH 8.0, 25°C Colossoma macropomum
8.32
-
Abz-RHFK-EDDnp pH 8.0, 25°C Colossoma macropomum
8.87
-
Abz-FRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
9.15
-
Abz-RGFK-EDDnp pH 8.0, 25°C Colossoma macropomum
10.65
-
Abz-RYFK-EDDnp pH 8.0, 25°C Colossoma macropomum
12.64
-
Abz-QRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
13.31
-
Abz-RQFK-EDDnp pH 8.0, 25°C Colossoma macropomum
16.63
-
Abz-RSFK-EDDnp pH 8.0, 25°C Colossoma macropomum
16.63
-
Abz-RNFK-EDDnp pH 8.0, 25°C Colossoma macropomum
17.3
-
Abz-ERFK-EDDnp pH 8.0, 25°C Colossoma macropomum
18.3
-
Abz-RTFK-EDDnp pH 8.0, 25°C Colossoma macropomum
21.07
-
Abz-HRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
26.61
-
Abz-RRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
27.72
-
Abz-YRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
30.61
-
Abz-NRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
31.94
-
Abz-GRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
46.57
-
Abz-VRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
53.23
-
Abz-LRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
53.23
-
Abz-KRFK-EDDnp pH 8.0, 25°C Colossoma macropomum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5 11.5 with substrate N-benzoyl-DL-arginine 4-nitroanilide Colossoma macropomum
9
-
with substrate N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin Colossoma macropomum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
30
-
Abz-RPFK-EDDnp pH 8.0, 25°C Colossoma macropomum
250
-
Abz-RVFK-EDDnp pH 8.0, 25°C Colossoma macropomum
290
-
Abz-RDFK-EDDnp pH 8.0, 25°C Colossoma macropomum
570
-
Abz-RGFK-EDDnp pH 8.0, 25°C Colossoma macropomum
580
-
Abz-REFK-EDDnp pH 8.0, 25°C Colossoma macropomum
1420
-
Abz-QRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
1800
-
Abz-RQFK-EDDnp pH 8.0, 25°C Colossoma macropomum
1940
-
Abz-WRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
1960
-
Abz-GRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
2520
-
Abz-ERFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3110
-
Abz-HRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3380
-
Abz-RNFK-EDDnp pH 8.0, 25°C Colossoma macropomum
4120
-
Abz-NRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
4490
-
Abz-RHFK-EDDnp pH 8.0, 25°C Colossoma macropomum
4980
-
Abz-YRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
5630
-
Abz-RTFK-EDDnp pH 8.0, 25°C Colossoma macropomum
6400
-
Abz-RYFK-EDDnp pH 8.0, 25°C Colossoma macropomum
16220
-
Abz-VRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
16300
-
Abz-RSFK-EDDnp pH 8.0, 25°C Colossoma macropomum
16460
-
Abz-RRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
16680
-
Abz-FRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
52600
-
Abz-LRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
91580
-
Abz-KRFK-EDDnp pH 8.0, 25°C Colossoma macropomum