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Literature summary for 3.1.1.74 extracted from
- Production optimization and characterization of recombinant cutinases from Thermobifida fusca sp. NRRL B-8184 (2013), Appl. Biochem. Biotechnol., 170, 654-675.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| DMF | activates 10% at 40% v/v | Thermobifida fusca |  |
| DMF | activates 6% at 40% v/v | Thermobifida fusca | |
| DMSO | activates 14% at 40% v/v | Thermobifida fusca | |
| DMSO | activates 9% at 40% v/v | Thermobifida fusca | |
| sodium 2-{[(3alpha,5beta,12alpha)-3,12-dihydroxy-24-oxocholan-24-yl]amino}ethane-1-sulfonate | activates 10% at 10 mM | Thermobifida fusca | |
| sodium 2-{[(3alpha,5beta,12alpha)-3,12-dihydroxy-24-oxocholan-24-yl]amino}ethane-1-sulfonate | activates 24% at 10 mM | Thermobifida fusca | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene cut1, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3), secretion into periplasm and medium, method development and optimization for process for large-scale production of cutinase in soluble form | Thermobifida fusca |
| gene cut2, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3), secretion into periplasm and medium, method development and optimization for process for large-scale production of cutinase in soluble form | Thermobifida fusca |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 1-Heptanol | 27% inhibition at 40% v/v; 28% inhibition at 40% v/v | Thermobifida fusca | |
| 1-Hexanol | 68% inhibition at 40% v/v; 86% inhibition at 40% v/v | Thermobifida fusca | |
| AgNO3 | 90% inhibition at 1 mM; 93% inhibition at 1 mM | Thermobifida fusca | |
| BaCl2 | 14% inhibition at 1 mM; 8% inhibition at 1 mM | Thermobifida fusca | |
| Benzene | 77% inhibition at 40% v/v; 79% inhibition at 40% v/v | Thermobifida fusca | |
| butanol | 10% inhibition at 40% v/v; 17% inhibition at 40% v/v | Thermobifida fusca | |
| CrCl2 | 99% inhibition at 1 mM; 99% inhibition at 1 mM | Thermobifida fusca | |
| CuSO4 | 26% inhibition at 1 mM; 32% inhibition at 1 mM | Thermobifida fusca | |
| deoxycholate | 24% inhibition at 10 mM; 25% inhibition at 10 mM | Thermobifida fusca | |
| Dichloromethane | 61% inhibition at 40% v/v; 74% inhibition at 40% v/v | Thermobifida fusca | |
| diethyldicarbonate | 54% inhibition at 1 mM; 56% inhibition at 1 mM | Thermobifida fusca | |
| FeSO4 | 55% inhibition at 1 mM; 55% inhibition at 1 mM | Thermobifida fusca | |
| hexyl acetate | 63% inhibition at 40% v/v; 83% inhibition at 40% v/v | Thermobifida fusca | |
| HgCl2 | 99% inhibition at 1 mM; 99% inhibition at 1 mM | Thermobifida fusca | |
| methanol | 8% inhibition at 40% v/v | Thermobifida fusca | |
| MgCl2 | 15% inhibition at 1 mM; 8% inhibition at 1 mM | Thermobifida fusca | |
| additional information | no or poor inhibition by tetrahydrofuran, n-hexane, methanol, ethanol, acetone, and acetonitrile at 40% v/v, or by 10 mM sodium deoxycholate and 1 mM EDTA; no or poor inhibition by tetrahydrofuran, triethylamine, ethanol, acetone, and acetonitrile at 40% v/v, or by 10 mM sodium deoxycholate and 1 mM EDTA | Thermobifida fusca | |
| n-hexane | 10% inhibition at 40% v/v | Thermobifida fusca | |
| PbCl2 | 53% inhibition at 1 mM; 54% inhibition at 1 mM | Thermobifida fusca | |
| PMSF | 48% inhibition at 1 mM; 59% inhibition at 1 mM | Thermobifida fusca | |
| RbCl | 93% inhibition at 1 mM; 95% inhibition at 1 mM | Thermobifida fusca | |
| SDS | 30% inhibition at 1 mM; 9% inhibition at 1 mM | Thermobifida fusca | |
| triethylamine | 24% inhibition at 40% v/v | Thermobifida fusca | |
| Triton X-100 | 35% inhibition at 1 mM; 55% inhibition at 1 mM | Thermobifida fusca | |
| Tween-20 | 28% inhibition at 1 mM; 35% inhibition at 1 mM | Thermobifida fusca | |
| Tween-80 | 24% inhibition at 1 mM; 60% inhibition at 1 mM | Thermobifida fusca | |
| Urea | 17% inhibition at 1 mM; 30% inhibition at 1 mM | Thermobifida fusca | |
| ZnSO4 | 59% inhibition at 1 mM; 65% inhibition at 1 mM | Thermobifida fusca | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Thermobifida fusca |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| KCl | activates 12% at 1 mM | Thermobifida fusca | |
| additional information | no effect on enzyme activity by 1 mM CoCl2, KCl, NiCl2, MgSO4, and MnCl2 | Thermobifida fusca | |
| additional information | no effect on enzyme activity by 1 mM CoCl2, MnCl2, and MgSO4 | Thermobifida fusca | |
| NaCl | activates 15% at 1 mM | Thermobifida fusca | |
| NaCl | activates 20% at 1 mM | Thermobifida fusca | |
| NiCl2 | activates 11% at 1 mM | Thermobifida fusca | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| cutin + H2O | Thermobifida fusca | - |
cutin monomers | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermobifida fusca | G8GER6 | gene Cut1 | - |
| Thermobifida fusca | Q6A0I4 | gene cut2 | - |
| Thermobifida fusca NRRL B-8184 | G8GER6 | gene Cut1 | - |
| Thermobifida fusca NRRL B-8184 | Q6A0I4 | gene cut2 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) culture supernatant by nickel affinity chromatography to homogeneity | Thermobifida fusca |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.218 | - |
pH 8.0, 50°C, purified recombinant His-tagged enzyme, 4-nitrophenyl (16-methyl sulfoyl ester) hexadecanoate | Thermobifida fusca |
| 0.395 | - |
pH 8.0, 50°C, purified recombinant His-tagged enzyme, 4-nitrophenyl (16-methyl sulfone ester) hexadecanoate | Thermobifida fusca |
| 542.5 | - |
pH 8.0, 50°C, purified recombinant His-tagged enzyme, substrate 4-nitrophenyl butyrate | Thermobifida fusca |
| 643.4 | - |
pH 8.0, 50°C, purified recombinant His-tagged enzyme, substrate 4-nitrophenyl butyrate | Thermobifida fusca |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl (16-methyl sulfone ester) hexadecanoate + H2O | - |
Thermobifida fusca | ? | - |
? | |
| 4-nitrophenyl (16-methyl sulfonyl ester) hexadecanoate + H2O | - |
Thermobifida fusca | 4-nitrophenol + (16-methyl sulfonyl ester) hexadecanoate | - |
? | |
| 4-nitrophenyl acetate + H2O | high activity | Thermobifida fusca | 4-nitrophenol + acetate | - |
? | |
| 4-nitrophenyl acetate + H2O | high activity | Thermobifida fusca NRRL B-8184 | 4-nitrophenol + acetate | - |
? | |
| 4-nitrophenyl butyrate + H2O | best 4-nitrophenyl ester substrate | Thermobifida fusca | 4-nitrophenol + butyrate | - |
? | |
| 4-nitrophenyl butyrate + H2O | best 4-nitrophenyl ester substrate | Thermobifida fusca NRRL B-8184 | 4-nitrophenol + butyrate | - |
? | |
| 4-nitrophenyl decanoate + H2O | - |
Thermobifida fusca | 4-nitrophenol + decanoate | - |
? | |
| 4-nitrophenyl decanoate + H2O | - |
Thermobifida fusca NRRL B-8184 | 4-nitrophenol + decanoate | - |
? | |
| 4-nitrophenyl laurate + H2O | - |
Thermobifida fusca | 4-nitrophenol + laurate | - |
? | |
| 4-nitrophenyl laurate + H2O | low activity | Thermobifida fusca | 4-nitrophenol + laurate | - |
? | |
| 4-nitrophenyl myristate + H2O | low activity | Thermobifida fusca | 4-nitrophenol + myristate | - |
? | |
| 4-nitrophenyl octanoate + H2O | - |
Thermobifida fusca | 4-nitrophenol + octanoate | - |
? | |
| 4-nitrophenyl octanoate + H2O | - |
Thermobifida fusca NRRL B-8184 | 4-nitrophenol + octanoate | - |
? | |
| 4-nitrophenyl palmitate + H2O | low activity | Thermobifida fusca | 4-nitrophenol + palmitate | - |
? | |
| 4-nitrophenyl valerate + H2O | - |
Thermobifida fusca | 4-nitrophenol + valerate | - |
? | |
| 4-nitrophenyl valerate + H2O | - |
Thermobifida fusca NRRL B-8184 | 4-nitrophenol + valerate | - |
? | |
| cutin + H2O | - |
Thermobifida fusca | cutin monomers | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Cut1 | - |
Thermobifida fusca |
| Cut2 | - |
Thermobifida fusca |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
- |
Thermobifida fusca |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | 80 | activity range | Thermobifida fusca |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
purified recombinant enzyme, residual activity is over 75% after 125 h | Thermobifida fusca |
| 55 | - |
purified recombinant enzyme, residual activity is over 38% after 125 h | Thermobifida fusca |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
- |
Thermobifida fusca |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6.8 | 9 | activity range | Thermobifida fusca |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1350 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C | Thermobifida fusca | |
| 2840 | - |
4-nitrophenyl butyrate | pH 8.0, 50°C | Thermobifida fusca | |
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