Cloned (Comment) | Organism |
---|---|
recombinant expression of extracellular wild-type and mutant enzymes in Escherichia coli | uncultured bacterium |
Crystallization (Comment) | Organism |
---|---|
purified recombinant C275A/C292A mutant enzyme, sitting drop vapor diffusion method, from 20% w/v PEG 3350 and 0.2 M sodium thiocyanate, 20°C, 2 weeks, X-ray diffraction structure determination and analysis at 1.5 A resolution | uncultured bacterium |
Protein Variants | Comment | Organism |
---|---|---|
C275A/C292A | site-directed mutagenesis, the mutant lacks the disulfide bond formed by Cys275 and Cys292, resulting in increased instability | uncultured bacterium |
General Stability | Organism |
---|---|
the disulfide bond formed by Cys275 and Cys292 contributes not only to the thermodynamic stability but also to the kinetic stability of LC-cutinase | uncultured bacterium |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
guanidine hydrochloride | GdnHCl-induced unfolding of LC-cutinase is analyzed at pH 8.0 by circular dichroism spectroscopy, overview | uncultured bacterium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
0.19 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
0.21 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
0.22 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
0.24 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
0.25 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
0.25 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
0.27 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant wild-type enzyme, presence of 1% PEG | uncultured bacterium | |
0.27 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant wild-type enzyme, presence of 1% PEG | uncultured bacterium |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
uncultured bacterium | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | uncultured bacterium | the enzyme shows polyethylene terephthalate-degrading activity | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
uncultured bacterium | - |
metagenome-derived LC-cutinase | - |
Purification (Comment) | Organism |
---|---|
recombinant extracellular wild-type and mutant enzymes without a putative N-terminal signal peptide (Met1-Ala34) and Gln35 from Escherichia coli | uncultured bacterium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl butyrate + H2O | - |
uncultured bacterium | 4-nitrophenol + butyrate | - |
? | |
additional information | the enzyme shows polyethylene terephthalate-degrading activity | uncultured bacterium | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LC-cutinase | - |
uncultured bacterium |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
uncultured bacterium |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 70 | activity range | uncultured bacterium |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermal denaturation-induced unfolding of LC-cutinase is analyzed at pH 8.0 by circular dichroism spectroscopy, kinetics of transition of the thermal denaturation, overview. The disulfide bond formed by Cys275 and Cys292 contributes not only to the thermodynamic stability but also to the kinetic stability of LC-cutinase | uncultured bacterium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
49 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
196 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
213 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
232 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
278 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant wild-type enzyme, presence of 1% PEG | uncultured bacterium | |
318 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
342 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
343 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
359 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant wild-type enzyme, presence of 1% PEG | uncultured bacterium |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | uncultured bacterium |
General Information | Comment | Organism |
---|---|---|
additional information | residues Ser165, Asp210, and His242 form the catalytic triad. The disulfide bond formed by Cys275 and Cys292 contributes not only to the thermodynamic stability but also to the kinetic stability of LC-cutinase | uncultured bacterium |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
258 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
888 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
1046 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant wild-type enzyme, presence of 1% PEG | uncultured bacterium | |
1050 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
1090 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
1295 | - |
4-nitrophenyl butyrate | pH 8.0, 70°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium | |
1311 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant wild-type enzyme, presence of 1% PEG | uncultured bacterium | |
1370 | - |
4-nitrophenyl butyrate | pH 8.0, 30°C, recombinant mutant C275A/C292A, absence of 1% PEG | uncultured bacterium | |
1630 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C, recombinant wild-type enzyme, absence of 1% PEG | uncultured bacterium |