Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | at 1% | uncultured organism | |
DMSO | increase in enzyme activity (130%) is observed at 25% v/v DMSO although at a higher concentration (50%), DMSO decreases enzyme activity to 72% | uncultured organism | |
DTT | at 1% | uncultured organism |
Cloned (Comment) | Organism |
---|---|
gene est06, screening of a forest soil metagenomic library, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, esterase (Est06) belonging to the hormone- sensitive lipase family of lipolytic enzymes is subcloned and heterologously expressed in Escherichia coli strain BL21(DE3) | uncultured organism |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 45% inhibition at 1 mM | uncultured organism | |
DMSO | increase in enzyme activity (130%) is observed at 25% v/v DMSO although at a higher concentration (50%), DMSO decreases enzyme activity to 72% | uncultured organism | |
Fe2+ | 35% inhibition at 1 mM | uncultured organism | |
additional information | no inhibition by EDTA, poor effect by Triton X-100 | uncultured organism | |
PMSF | strong inhibition | uncultured organism | |
SDS | complete inactivation | uncultured organism | |
Tween 20 | - |
uncultured organism | |
Tween 80 | - |
uncultured organism |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
NaCl | moderate stability with NaCl up to 2.5 M with the residual activity being over 75%, 1 h, 25°C | uncultured organism |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
uncultured organism | G3CRD1 | a forest soil metagenome-derived gene | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
5.76 | - |
purified recombinant enzyme, pH 7.0, 30°C, substrate 4-nitrophenyl valerate | uncultured organism |
Storage Stability | Organism |
---|---|
10-40°C, purified recombinant enzyme Est06 retains all its activity at 10, 25 and 30°C over the entire period. After six days of incubation at 40°C, esterase activity remains above 70%, considerable loss in Est06 activity (less than 65% residual activity) is observed only after the 10th day of incubation | uncultured organism |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl acetate + H2O | high activity | uncultured organism | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl butyrate + H2O | high activity | uncultured organism | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl caprate + H2O | low activity | uncultured organism | 4-nitrophenol + caprate | - |
? | |
4-nitrophenyl caproate + H2O | - |
uncultured organism | 4-nitrophenol + caproate | - |
? | |
4-nitrophenyl caprylate + H2O | low activity | uncultured organism | 4-nitrophenol + caprylate | - |
? | |
4-nitrophenyl laurate + H2O | low activity | uncultured organism | 4-nitrophenol + laurate | - |
? | |
4-nitrophenyl myristate + H2O | low activity | uncultured organism | 4-nitrophenol + myristate | - |
? | |
4-nitrophenyl palmitate + H2O | low activity | uncultured organism | 4-nitrophenol + palmitate | - |
? | |
4-nitrophenyl valerate + H2O | best substrate | uncultured organism | 4-nitrophenol + valerate | - |
? | |
additional information | the purified enzyme Est06 exhibits high affinity for acyl esters with short-chain fatty acids, and shows optimum activity with p-nitrophenyl valerate | uncultured organism | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 31000, about, sequence calculation, x* 35000, recombinant enzyme, SDS-PAGE | uncultured organism |
More | tertiary structure prediction confirmed the presence of two alpha-helices, alpha1 and alpha2, that constitute the N-terminal cap domain followed by the alpha/beta fold of the catalytic domain. The hydrolase fold comprises eight beta-strands that form parallel structures, with Ser144 located between beta5 and alpha5, Glu238 between beta7 and alpha7 and His268 between beta8 and alpha9. Two cysteine residues, Cys46 and Cys170, which may participate in disulphide bridge formation | uncultured organism |
Synonyms | Comment | Organism |
---|---|---|
Est06 | - |
uncultured organism |
family IV carboxylesterase | - |
uncultured organism |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
uncultured organism |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
purified recombinant enzyme Est06 exhibits high stability at moderate temperatures by retaining all of its catalytic activity below 30°C over 13 days | uncultured organism |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
uncultured organism |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 9.5 | over 50% of maximal activity within this range | uncultured organism |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5 | 9 | purified recombinant enzyme Est06 displays high stability within this range, at 25°C for 4 h | uncultured organism |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
uncultured organism | sequence calculation | - |
5.99 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a metagenome-derived member of the hormone-sensitive lipase family of lipolytic enzymes | uncultured organism |
additional information | Est06 has a HGGG motif (residues 74 to 77) and a catalytic triad composed of Ser144, Glu238, and His268 | uncultured organism |