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Literature summary for 3.1.1.74 extracted from
- Thermal stability engineering of Glomerella cingulata cutinase (2013), Protein Eng. Des. Sel., 26, 369-375.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| sequence comparisons, functional recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Origami B (DE3) | Colletotrichum gloeosporioides |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| L172K | site-directed mutagenesis, compared to the wild-type enzyme, the mutant exhibits higher enzymatic performance towards phenyl ester substrates of longer carbon chain length, yet its thermal stability is inversely affected | Colletotrichum gloeosporioides |
| N177D | site-directed mutagenesis, the mutation aims to alter the surface electrostatics as well as to remove a potentially deamidation-prone asparagine residue. The mutant is more resilient to temperature increase with a 2.7fold increase in half-life at 50°C, accompanied by an increase in optimal temperature, as compared with wild-type enzyme, while the activity at 25°C is not compromised | Colletotrichum gloeosporioides |
| N177D/L172K | site-directed mutagenesis, the double mutant shows enhanced activity towards phenyl ester substrates and enhanced thermal stability | Colletotrichum gloeosporioides |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 39000 | - |
x * 39000, recombinant His-tagged enzyme, SDS-PAGE | Colletotrichum gloeosporioides |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Colletotrichum gloeosporioides | P11373 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Origami B (DE3) by nickel affinity chromatography, tag cleavage by recombinant enterokinase, and anion exchange chromatography | Colletotrichum gloeosporioides |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl laurate + H2O | - |
Colletotrichum gloeosporioides | 4-nitrophenol + laurate | - |
? |  |
| 4-nitrophenyl myristate + H2O | - |
Colletotrichum gloeosporioides | 4-nitrophenol + myristate | - |
? | |
| 4-nitrophenyl palmitate + H2O | - |
Colletotrichum gloeosporioides | 4-nitrophenol + palmitate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 39000, recombinant His-tagged enzyme, SDS-PAGE | Colletotrichum gloeosporioides |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
recombinant wild-type enzyme | Colletotrichum gloeosporioides |
| 30 | - |
recombinant mutant N177D | Colletotrichum gloeosporioides |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
purified wild-type enzyme and mutant L172K retain 10% activity after 2 h, mutant N177D retains 30% activity after 2 h with half-life of 30 min | Colletotrichum gloeosporioides |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | residues N168, Q170 an N171 of Glomerella cingulata are highly conserved with all cutinases of fungal phytopathogens | Colletotrichum gloeosporioides |
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Release 2023.1 (January 2023)
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