Cloned (Comment) | Organism |
---|---|
library screening, DNA and amino acid sequence determination and anaylsis, expression of wild-type and mutant enzymes in Pichia pastoris strain GS115 as secreted proteins, subcloning in Escherichia coli | Rhizopus arrhizus |
Protein Variants | Comment | Organism |
---|---|---|
A9T | random mutagenesis, the mutation of residue A9 results in only marginally affected thermostability and optimum reaction temperature compared to the wild-type enzyme | Rhizopus arrhizus |
A9T/E190V/M225I | random mutagenesis, the mutation of residues A9 and M225 result in only marginally affected thermostability and optimum reaction temperature compared to the wild-type enzyme, while the mutation of E190 is critical for thermostability and optimum reaction temperature | Rhizopus arrhizus |
E190V | random mutagenesis, the mutation of E190 is critical for thermostability and optimum reaction temperature | Rhizopus arrhizus |
M225I | random mutagenesis, the mutation of residues M225 result in only marginally affected thermostability and optimum reaction temperature compared to the wild-type enzyme | Rhizopus arrhizus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhizopus arrhizus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
secreted recombinant wild-type and mutant enzymes from Pichia pastoris strain GS115 by ultrafiltration, and dialysis, followed by anion exchange and hydrophobic interaction chromatography | Rhizopus arrhizus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
triacylglycerol + H2O = diacylglycerol + a carboxylate | the catalytic center is formed by Ser145, Asp204, His257 | Rhizopus arrhizus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2050 | - |
purified recombinant mutant enzyme | Rhizopus arrhizus |
2500 | - |
purified recombinant wild-type enzyme | Rhizopus arrhizus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
olive oil + H2O | - |
Rhizopus arrhizus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
lipase | - |
Rhizopus arrhizus |
RAL | - |
Rhizopus arrhizus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
- |
Rhizopus arrhizus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 60 | mutant enzyme | Rhizopus arrhizus |
30 | 50 | wild-type enzyme | Rhizopus arrhizus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
the half-life of the purified recombinant wild-type enzyme is 7 min, for the mutant A9T/E190V/M225I 83 min, for mutant A9T 7 min, for mutant M225I 9 min, and for mutant E190V 76 min | Rhizopus arrhizus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Rhizopus arrhizus |