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Literature summary for 3.1.1.3 extracted from

  • Chang, S.W.; Lee, G.C.; Shaw, J.F.
    Efficient production of active recombinant Candida rugosa LIP3 lipase in Pichia pastoris and biochemical characterization of the purified enzyme (2006), J. Agric. Food Chem., 54, 5831-5838.
    View publication on PubMed

Activating Compound

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(Commentary)

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

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

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

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

Organism UniProt Comment Textmining
Diutina rugosa P32947 LIP3 precursor; i.e. Candida cylindracea, gene lip3, isozyme LIP3
-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl butyrate + H2O
-
Diutina rugosa 4-nitrophenol + butyrate
-
?

Synonyms

Synonyms Comment Organism
Candida rugosa lipase
-
Diutina rugosa
CRL
-
Diutina rugosa
LIP3
-
Diutina rugosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Diutina rugosa

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
10 90
-
Diutina rugosa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37 90 10 min Diutina rugosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4 6
-
Diutina rugosa

pH Range

pH Minimum pH Maximum Comment Organism
3 9 inactive at pH 10.0, pH profile, overview Diutina rugosa

pH Stability

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