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Literature summary for 3.1.1.74 extracted from

  • Ribitsch, D.; Herrero Acero, E.; Przylucka, A.; Zitzenbacher, S.; Marold, A.; Gamerith, C.; Tscheliessnig, R.; Jungbauer, A.; Rennhofer, H.; Lichtenegger, H.; Amenitsch, H.; Bonazza, K.; Kubicek, C.P.; Druzhinina, I.S.; Guebitz, G.M.
    Enhanced cutinase-catalyzed hydrolysis of polyethylene terephthalate by covalent fusion to hydrophobins (2015), Appl. Environ. Microbiol., 81, 3586-3592 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.1.74

Application

Application Comment Organism
degradation fusion of enzyme to the class II hydrophobins HFB4 and HFB7 or the pseudo-class I hydrophobin HFB9b. Upon fusion to HFB4 or HFB7, the hydrolysis of polyethylene terephthalate is enhanced over16fold over the level with the free enzyme Thermobifida cellulosilytica

Protein Variants

Protein Variants Comment Organism
additional information fusion of enzyme to the class II hydrophobins HFB4 and HFB7 or the pseudo-class I hydrophobin HFB9b. The fusion enzymes exhibit decreased kcat values on soluble substrates and strongly decreased the hydrophilicity of glass but cause only small changes in the hydrophobicity of polyethylene terephthalate. Upon fusion to HFB4 or HFB7, the hydrolysis of polyethylene terephthalate is enhanced over16fold over the level with the free enzyme. Fusion with the non-class II hydrophobin HFB9b does not increase the rate of hydrolysis over that of the enzyme-hydrophobin mixture, but HFB9b performs best when polyethylene terephthalate is preincubated with the hydrophobins before enzyme treatment. The pattern of hydrolysis by the fusion shows an increase in the concentration of the product mono(2-hydroxyethyl) terephthalate relative to that of the main product, terephthalic acid Thermobifida cellulosilytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.8
-
 4-nitrophenyl butanoate wild-type, pH 7.0, 25°C Thermobifida cellulosilytica
0.8
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB9b, pH 7.0, 25°C Thermobifida cellulosilytica
0.9
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB7, pH 7.0, 25°C Thermobifida cellulosilytica
1.3
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB4, pH 7.0, 25°C Thermobifida cellulosilytica
1.4
-
 4-nitrophenyl acetate wild-type, pH 7.0, 25°C Thermobifida cellulosilytica
1.5
-
 4-nitrophenyl acetate fusion protein Cut1-HFB9b, pH 7.0, 25°C Thermobifida cellulosilytica
2.3
-
 4-nitrophenyl acetate fusion protein Cut1-HFB7, pH 7.0, 25°C Thermobifida cellulosilytica
2.4
-
 4-nitrophenyl acetate fusion protein Cut1-HFB4, pH 7.0, 25°C Thermobifida cellulosilytica

Organism

Organism UniProt Comment Textmining
Thermobifida cellulosilytica E9LVH8 isoform Cut1
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O
-
 Thermobifida cellulosilytica 4-nitrophenol + acetate
-
 ?
4-nitrophenyl butanoate + H2O
-
 Thermobifida cellulosilytica 4-nitrophenol + butanoate
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
49
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB9b, pH 7.0, 25°C Thermobifida cellulosilytica
86
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB4, pH 7.0, 25°C Thermobifida cellulosilytica
99
-
 4-nitrophenyl acetate fusion protein Cut1-HFB4, pH 7.0, 25°C Thermobifida cellulosilytica
105
-
 4-nitrophenyl acetate fusion protein Cut1-HFB9b, pH 7.0, 25°C Thermobifida cellulosilytica
112
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB7, pH 7.0, 25°C Thermobifida cellulosilytica
129
-
 4-nitrophenyl acetate fusion protein Cut1-HFB7, pH 7.0, 25°C Thermobifida cellulosilytica
325
-
 4-nitrophenyl butanoate wild-type, pH 7.0, 25°C Thermobifida cellulosilytica
363
-
 4-nitrophenyl acetate wild-type, pH 7.0, 25°C Thermobifida cellulosilytica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
42
-
 4-nitrophenyl acetate fusion protein Cut1-HFB4, pH 7.0, 25°C Thermobifida cellulosilytica
55
-
 4-nitrophenyl acetate fusion protein Cut1-HFB7, pH 7.0, 25°C Thermobifida cellulosilytica
62
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB9b, pH 7.0, 25°C Thermobifida cellulosilytica
68
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB4, pH 7.0, 25°C Thermobifida cellulosilytica
71
-
 4-nitrophenyl acetate fusion protein Cut1-HFB9b, pH 7.0, 25°C Thermobifida cellulosilytica
130
-
 4-nitrophenyl butanoate fusion protein Cut1-HFB7, pH 7.0, 25°C Thermobifida cellulosilytica
291
-
 4-nitrophenyl acetate wild-type, pH 7.0, 25°C Thermobifida cellulosilytica
406
-
 4-nitrophenyl butanoate wild-type, pH 7.0, 25°C Thermobifida cellulosilytica