Cloned (Comment) | Organism |
---|---|
the enzyme is cloned and expressed in Escherichia coli BL21(DE3) and Pichia pastoris GS115 cells without a signal peptide sequence | Passiflora edulis |
Protein Variants | Comment | Organism |
---|---|---|
N105Q | the mutant enzyme shows much lower thermostability, pH stability, and organic solvent tolerance than the hydroxynitrile lyase from leaves and that expressed in Pichia pastoris. kcat of the mutant enzyme is 1.3fold lower than the kcat of the wild-type enzyme, kcat/Km of the mutant enzyme is 2.3fold lower than the kcat/Km of the wild type enzyme from leaves | Passiflora edulis |
General Stability | Organism |
---|---|
the presence of glycan can prevent the self-degradation of this enzyme at 10°C | Passiflora edulis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11.2 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase from leaves | Passiflora edulis | |
13 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase expressed in Pichia pastoris | Passiflora edulis | |
19.8 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris | Passiflora edulis | |
26.1 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase expressed in Escherichia coli | Passiflora edulis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
13000 | - |
hydroxynitrile lyase expressed in Escherichia coli, SDS-PAGE | Passiflora edulis |
13000 | - |
hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris, SDS-PAGE | Passiflora edulis |
14000 | - |
hydroxynitrile lyase from leaves, SDS-PAGE | Passiflora edulis |
14067 | - |
calculated from sequence, mature protein without signal peptide | Passiflora edulis |
17000 | - |
hydroxynitrile lyase expressed in Pichia pastoris, SDS-PAGE | Passiflora edulis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Passiflora edulis | A0A1L7NZN4 | var. flavicarpa | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | hydroxynitrile lyase from leaves and that expressed in Pichia pastoris are glycosylated, whereas that expressed in Escherichia coli is not. Predicted mature enzyme has a N-glycosylation site on the asparagine residue at 105 | Passiflora edulis |
Purification (Comment) | Organism |
---|---|
- |
Passiflora edulis |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Passiflora edulis | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
54.1 | - |
25°C, pH 4, hydroxynitrile lyase expressed in Escherichia coli | Passiflora edulis |
133.6 | - |
25°C, pH 4, hydroxynitrile lyase from leaves | Passiflora edulis |
134 | - |
25°C, pH 4, hydroxynitrile lyase expressed in Pichia pastoris | Passiflora edulis |
140 | - |
25°C, pH 4, hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris | Passiflora edulis |
Storage Stability | Organism |
---|---|
10°C, 12 h, the stabilities of hydroxynitrile lyase from leaves and hydroxynitrile lyase expressed in Pichia pastoris are comparable, with remaining enzyme activities of 60-94%. The activities of hydroxynitrile lyase mutant N105Q expressed in Pichia pastor and hydroxynitrile lyase expressed in Escherichia coli vary between 70% and 0% in all biphasic systems apart from those with DEE and DIPE. The nonglycosylated hydroxynitrile lyase expressed in Escherichia coli and hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris both lose 50% of their activity in aqueous buffer with shaking at 1500 rpm at 10°C | Passiflora edulis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyanide + benzaldehyde | the glycosylated enzyme efficiently performs transcyanation of (R)-mandelonitrile with a 98% enantiomeric excess in a biphasic system with diisopropyl ether | Passiflora edulis | (R)-mandelonitrile | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
the hydroxynitrile lyase from leaves and that expressed in Pichia pastoris show much better thermostability, pH stability, and organic solvent tolerance than the deglycosylated enzyme expressed in Escherichia coli and the deglycosylated mutant N105Q expressed in Pichia pastoris | Passiflora edulis |
30 | 50 | 1 h, stable, hydroxynitrile lyase from leaves, hydroxynitrile lyase expressed in Pichia pastoris, hydroxynitrile lyase expressed in Escherichia coli, hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris | Passiflora edulis |
55 | - |
1 h, 10% loss of activity, hydroxynitrile lyase from leaves and hydroxynitrile lyase expressed in Pichia pastoris | Passiflora edulis |
60 | - |
1h, hydroxynitrile lyase from leaves loses 20% of its activity. Hydroxynitrile lyase expressed in Pichia pastoris loses 40% of its activity. Hydroxynitrile lyase expressed in Escherichia coli and hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris completely lose activity | Passiflora edulis |
70 | - |
1 h, hydroxynitrile lyase from leaves loses 60% of its activity, Hydroxynitrile lyase expressed in Pichia pastoris, completely loses activity | Passiflora edulis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
38.4 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyaseexpressed in Escherichia coli | Passiflora edulis | |
39.7 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris | Passiflora edulis | |
50.9 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase from leaves | Passiflora edulis | |
52.1 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase expressed in Pichia pastoris | Passiflora edulis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3.5 | 10 | activities of all the hydroxynitrile lyase are broadly stable over the range from pH 3.5 to 10.0, although the activity of hydroxynitrile lyase expressed in Escherichia coli is reduced to 60% and 70% at pH 7.0 and 10.0, respectively | Passiflora edulis |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Passiflora edulis | calculated from sequence, mature protein without signal peptide | - |
5.2 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.47 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase expressed in Escherichia coli | Passiflora edulis | |
2 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase mutant N105Q expressed in Pichia pastoris | Passiflora edulis | |
4 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase expressed in Pichia pastoris | Passiflora edulis | |
4.56 | - |
benzaldehyde | 25°C, pH 4, hydroxynitrile lyase from leaves | Passiflora edulis |