Cloned (Comment) | Organism |
---|---|
mutant enzymes S154L and S154L/S293L are expressed in Pichia pastoris, no obvious expression is observed from wild type and the S293L mutant enzymes | Candida albicans |
Protein Variants | Comment | Organism |
---|---|---|
S154L | the mutation results in loss of enzyme expression | Candida albicans |
S154L/S293L | the mutant exhibits a preference for the short- and medium-chain length 4-nitrophenyl (C4 and C8 acyl group) esters rather than the long chain length 4-nitrophenyl esters (C12, C16 and C18 acyl group), the pure Lip5 double mutant is stable at pH 5.0-9.0 and at temperatures ranging from 15-35°C | Candida albicans |
S293L | the mutation causes a marked reduction in the lipase activity | Candida albicans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ba2+ | 77.22% residual activity at 5 mM | Candida albicans | |
Ca2+ | 72.04% residual activity at 5 mM | Candida albicans | |
Cu2+ | 78.35% residual activity at 5 mM | Candida albicans | |
EDTA | 65.41% residual activity at 5 mM | Candida albicans | |
Fe2+ | strong inhibition with residual activity of 20.27% at 5 mM | Candida albicans | |
Fe3+ | strong inhibition with residual activity of 35.11% at 5 mM | Candida albicans | |
Hg2+ | strong inhibition with residual activity of 13.19% at 5 mM | Candida albicans | |
Li+ | 92.24% residual activity at 5 mM | Candida albicans | |
Mg2+ | 94.49% residual activity at 1 mM | Candida albicans | |
Mn2+ | 90.66% residual activity at 5 mM | Candida albicans | |
Ni2+ | 92.24% residual activity at 5 mM | Candida albicans | |
phenylmethylsulfonyl fluoride | 38.54% residual activity 5 mM | Candida albicans | |
SDS | SDS sharply decreases the lipase activity by 85% at 0.1% (w/v) | Candida albicans | |
Triton X-100 | 48.55% residual activity at 0.1% (v/v) | Candida albicans | |
Tween 20 | 82.52% residual activity at 0.1% (v/v) | Candida albicans | |
Tween 80 | 57.24% residual activity at 0.1% (v/v) | Candida albicans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.27 | - |
4-nitrophenyl caprylate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
0.36 | - |
4-nitrophenyl palmitate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
0.41 | - |
4-nitrophenyl butyrate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
0.5 | - |
4-nitrophenyl laurate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | Zn2+ increases the recombinant lipase activity at 1 mM to 119.22% or at 5 mM to 115.19% | Candida albicans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
50000 | - |
x * 50000, S154L and S154L/S293L mutant enzymes | Candida albicans |
Organic Solvent | Comment | Organism |
---|---|---|
Acetone | the residual activities of Lip5 double mutant remains at 44.76% after incubation for 1 h in 30% (v/v) acetone | Candida albicans |
Ethanol | the residual activities of Lip5 double mutant remains at 24.32% after incubation for 1 h in 30% (v/v) ethanol | Candida albicans |
isopropanol | lipase activity is almost lost (4.59% residual activity) in the presence of 30% (v/v) isopropanol | Candida albicans |
Methanol | the residual activities of Lip5 double mutant remains at 48.23% after incubation for 1 h in 30% (v/v) methanol | Candida albicans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candida albicans | - |
- |
- |
Candida albicans ATCC 10231 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
ammonium sulfate precipitation and Q Sepharose column chromatography | Candida albicans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl butyrate + H2O | - |
Candida albicans | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Candida albicans ATCC 10231 | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl caprylate + H2O | highest activity of mutant enzyme S154L/S293L | Candida albicans | 4-nitrophenol + caprylate | - |
? | |
4-nitrophenyl caprylate + H2O | highest activity of mutant enzyme S154L/S293L | Candida albicans ATCC 10231 | 4-nitrophenol + caprylate | - |
? | |
4-nitrophenyl laurate + H2O | lowest activity of mutant enzyme S154L/S293L | Candida albicans | 4-nitrophenol + laurate | - |
? | |
4-nitrophenyl laurate + H2O | lowest activity of mutant enzyme S154L/S293L | Candida albicans ATCC 10231 | 4-nitrophenol + laurate | - |
? | |
4-nitrophenyl palmitate + H2O | - |
Candida albicans | 4-nitrophenol + palmitate | - |
? | |
4-nitrophenyl palmitate + H2O | - |
Candida albicans ATCC 10231 | 4-nitrophenol + palmitate | - |
? | |
additional information | the enzyme also uses olive oil and safflower oil as substrates | Candida albicans | ? | - |
? | |
additional information | the enzyme also uses olive oil and safflower oil as substrates | Candida albicans ATCC 10231 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 50000, S154L and S154L/S293L mutant enzymes | Candida albicans |
Synonyms | Comment | Organism |
---|---|---|
Lip5 | - |
Candida albicans |
lipase | - |
Candida albicans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 25 | the recombinant lipase displays the highest activity at 15 and 25°C | Candida albicans |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
5 | 25 | the recombinant lipase maintains more than 70% of the maximum activity at 35°C. Even at a lower temperature of 5°C, the recombinant lipase still retains more than 50% of the maximal activity | Candida albicans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 45 | the Lip5 double mutant shows good thermostability over a range of temperature from 15°C to 45°C, with more than 60% of maximal activity retained after 2 h incubation. However, the stability of recombinant lipase decreases at 55°C, with 23.86% residual activity after incubation for 2 h | Candida albicans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9.41 | - |
4-nitrophenyl laurate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
31.59 | - |
4-nitrophenyl palmitate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
54.18 | - |
4-nitrophenyl butyrate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
55.12 | - |
4-nitrophenyl caprylate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | 6 | the recombinant lipase displays the highest activity at pH 5.0 and 6.0 | Candida albicans |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4 | 9 | the Lip5-double mutant activity exhibits stable within the pH range of 5.0-9.0, and it remains more than 60% of maximal activity. The recombinant lipase sharply loses its activity when incubated at pH 4.0, and only preserves 26% activity after 12 h of incubation | Candida albicans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
18.58 | - |
4-nitrophenyl laurate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
86.86 | - |
4-nitrophenyl palmitate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
131 | - |
4-nitrophenyl butyrate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans | |
203.4 | - |
4-nitrophenyl caprylate | mutant enzyme S154L/S293L, at pH 6.0 and 25°C | Candida albicans |