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Literature summary 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.21.4 0.0005
-
 Abz-FRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0006
-
 Abz-KRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.001
-
 Abz-RSFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.001
-
 Abz-LRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0017
-
 Abz-RYFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0017
-
 Abz-RRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.00185
-
 Abz-RHFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0029
-
 Abz-VRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0031
-
 Abz-RPFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.00325
-
 Abz-RTFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0033
-
 Abz-WRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0049
-
 Abz-RNFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0056
-
 Abz-YRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0068
-
 Abz-HRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0069
-
 Abz-ERFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0074
-
 Abz-RQFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0074
-
 Abz-NRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0081
-
 Abz-RDFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0089
-
 Abz-QRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0129
-
 Abz-REFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0159
-
 Abz-RGFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0163
-
 Abz-GRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 0.0183
-
 Abz-RVFK-EDDnp pH 8.0, 25°C Colossoma macropomum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.21.4 23900
-
 x * 23900, SDS-PAGE Colossoma macropomum

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.4 Colossoma macropomum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.4 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.4 Abz-ERFK-EDDnp + H2O
-
 Colossoma macropomum Abz-ER + FK-EDDnp
-
 ?
3.4.21.4 Abz-FRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-FR + FK-EDDnp
-
 ?
3.4.21.4 Abz-GRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-GR + FK-EDDnp
-
 ?
3.4.21.4 Abz-HRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-HR + FK-EDDnp
-
 ?
3.4.21.4 Abz-KRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-KR + FK-EDDnp
-
 ?
3.4.21.4 Abz-LRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-LR + FK-EDDnp
-
 ?
3.4.21.4 Abz-NRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-NR + FK-EDDnp
-
 ?
3.4.21.4 Abz-QRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-QR + FK-EDDnp
-
 ?
3.4.21.4 Abz-RDFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + DFK-EDDnp
-
 ?
3.4.21.4 Abz-REFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + EFK-EDDnp
-
 ?
3.4.21.4 Abz-RGFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + GFK-EDDnp
-
 ?
3.4.21.4 Abz-RHFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + HFK-EDDnp
-
 ?
3.4.21.4 Abz-RNFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + NFK-EDDnp
-
 ?
3.4.21.4 Abz-RPFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + PFK-EDDnp
-
 ?
3.4.21.4 Abz-RQFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + QFK-EDDnp
-
 ?
3.4.21.4 Abz-RRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + RFK-EDDnp
-
 ?
3.4.21.4 Abz-RSFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + SFK-EDDnp
-
 ?
3.4.21.4 Abz-RTFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + TFK-EDDnp
-
 ?
3.4.21.4 Abz-RVFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + VFK-EDDnp
-
 ?
3.4.21.4 Abz-RYFK-EDDnp + H2O
-
 Colossoma macropomum Abz-R + YFK-EDDnp
-
 ?
3.4.21.4 Abz-VRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-VR + FK-EDDnp
-
 ?
3.4.21.4 Abz-WRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-WR + FK-EDDnp
-
 ?
3.4.21.4 Abz-YRFK-EDDnp + H2O
-
 Colossoma macropomum Abz-YR + FK-EDDnp
-
 ?
3.4.21.4 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 ?
-
 ?
3.4.21.4 N-benzoyl-DL-arginine 4-nitroanilide + H2O a synthetic trypsin substrate Colossoma macropomum N-benzoyl-DL-arginine + 4-nitroaniline
-
 ?
3.4.21.4 N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
-
 Colossoma macropomum N-benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
-
 ?

Subunits

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.21.4 50
-
 with substrate N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin Colossoma macropomum
3.4.21.4 70
-
 with substrate N-benzoyl-DL-arginine 4-nitroanilide Colossoma macropomum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.21.4 60
-
 1 h, purified enzyme, 20% activity remaining with substrate N-benzoyl-DL-arginine 4-nitroanilide Colossoma macropomum
3.4.21.4 60
-
 3 h, purified enzyme, 60% activity remaining with substrate N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin Colossoma macropomum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.21.4 0.1
-
 Abz-RPFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 2.33
-
 Abz-RDFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 4.59
-
 Abz-RVFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 6.32
-
 Abz-WRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 7.49
-
 Abz-REFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 8.32
-
 Abz-RHFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 8.87
-
 Abz-FRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 9.15
-
 Abz-RGFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 10.65
-
 Abz-RYFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 12.64
-
 Abz-QRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 13.31
-
 Abz-RQFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 16.63
-
 Abz-RSFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 16.63
-
 Abz-RNFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 17.3
-
 Abz-ERFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 18.3
-
 Abz-RTFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 21.07
-
 Abz-HRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 26.61
-
 Abz-RRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 27.72
-
 Abz-YRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 30.61
-
 Abz-NRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 31.94
-
 Abz-GRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 46.57
-
 Abz-VRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 53.23
-
 Abz-LRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 53.23
-
 Abz-KRFK-EDDnp pH 8.0, 25°C Colossoma macropomum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.21.4 7.5 11.5 with substrate N-benzoyl-DL-arginine 4-nitroanilide Colossoma macropomum
3.4.21.4 9
-
 with substrate N-benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin Colossoma macropomum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.21.4 30
-
 Abz-RPFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 250
-
 Abz-RVFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 290
-
 Abz-RDFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 570
-
 Abz-RGFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 580
-
 Abz-REFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 1420
-
 Abz-QRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 1800
-
 Abz-RQFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 1940
-
 Abz-WRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 1960
-
 Abz-GRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 2520
-
 Abz-ERFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 3110
-
 Abz-HRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 3380
-
 Abz-RNFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 4120
-
 Abz-NRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 4490
-
 Abz-RHFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 4980
-
 Abz-YRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 5630
-
 Abz-RTFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 6400
-
 Abz-RYFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 16220
-
 Abz-VRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 16300
-
 Abz-RSFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 16460
-
 Abz-RRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 16680
-
 Abz-FRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 52600
-
 Abz-LRFK-EDDnp pH 8.0, 25°C Colossoma macropomum
3.4.21.4 91580
-
 Abz-KRFK-EDDnp pH 8.0, 25°C Colossoma macropomum