Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Brij | slight inhibition of wild-type enzyme, and slight activation of the mutant N-fused enzyme at 1 mM | Diutina rugosa | |
sodium taurocholate | 55% inhibition of wild-type enzyme and slight activation of the mutant N-fused enzyme at 1 mM | Diutina rugosa |
Cloned (Comment) | Organism |
---|---|
gene lip3, subcloning in Escherichia coli strain DH5alpha, expression of wild-type enzyme and of genetically modified, active codon-optimized LIP3 lipase in Pichia pastoris strain SMD168H | Diutina rugosa |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the 18 nonuniversal serine codons of the lip3 gene are converted into universal serine codons by means of an overlap extension PCR-based multiple-site-directed mutagenesis to express an active recombinant LIP3 in Pichia pastoris, removal of additional N-terminal peptide in front of the lip3 gene by PCR, production yield of codon-optimized N-fused lip3 is 69fold increased compared to the non-codon-optimized enzyme | Diutina rugosa |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Brij | slight inhibition of wild-type enzyme, and slight activation of the mutant N-fused enzyme at 1 mM | Diutina rugosa | |
CHAPS | complete inhibition of both wild-type and mutant N-fused enzymes at 1 mM | Diutina rugosa | |
PMSF | about 40% inhibition at 10 mM, 20% at 1 mM | Diutina rugosa | |
SDS | complete inhibition of both wild-type and mutant N-fused enzymes at 0.1 mM | Diutina rugosa | |
sodium taurocholate | 55% inhibition of wild-type enzyme and slight activation of the mutant N-fused enzyme at 1 mM | Diutina rugosa | |
Triton X-100 | strong inhibition of both wild-type and mutant N-fused enzymes at 1 mM | Diutina rugosa | |
Tween 20 | over 90% inhibition of both wild-type and mutant N-fused enzymes at 1 mM | Diutina rugosa | |
Tween 80 | about 30% inhibition of both wild-type and mutant N-fused enzymes at 1 mM | Diutina rugosa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | 2.3fold activation at 10 mM | Diutina rugosa | |
additional information | the enzyme is not affected by Ca2+, EDTA, Mg2+, Ag+, ammmonium sulfate, NaCl, NaN3, and Cu2+ at 1-10 mM | Diutina rugosa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Diutina rugosa | P32947 | LIP3 precursor; i.e. Candida cylindracea, gene lip3, isozyme LIP3 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type LIP3 10.3fold and genetically modified, active codon-optimized LIP3 16.1fold from Pichia pastoris strain SMD168H | Diutina rugosa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
67.5 | - |
purified recombinant modified N-fused LIP3, substrate 4-nitrophenyl butyrate, pH 7.0, 37°C | Diutina rugosa |
82.6 | - |
purified recombinant wild-type LIP3, substrate 4-nitrophenyl butyrate, pH 7.0, 37°C | Diutina rugosa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl butyrate + H2O | - |
Diutina rugosa | 4-nitrophenol + butyrate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Candida rugosa lipase | - |
Diutina rugosa |
CRL | - |
Diutina rugosa |
LIP3 | - |
Diutina rugosa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Diutina rugosa |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
10 | 90 | - |
Diutina rugosa |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | 90 | 10 min | Diutina rugosa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4 | 6 | - |
Diutina rugosa |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | 9 | inactive at pH 10.0, pH profile, overview | Diutina rugosa |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3 | 9 | 16 h, 37°C, purified recombinant enzyme, 54% of maximal activity at pH 3.0, 52.5% at pH 9.0, inactivation at pH 10.0, completely stabel at pH 8.0, overview | Diutina rugosa |