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Literature summary for 1.10.3.2 extracted from

  • Autore, F.; Del Vecchio, C.; Fraternali, F.; Giardina, P.; Sannia, G.; Faraco, V.
    Molecular determinants of peculiar properties of a Pleurotus ostreatus laccase: analysis by site-directed mutagenesis (2009), Enzyme Microb. Technol., 45, 507-513.
No PubMed abstract available
Search for more literature for 1.10.3.2

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, subcloning in Escherichia coli strain Top10, expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain W303-1A Pleurotus ostreatus

Protein Variants

Protein Variants Comment Organism
D205R site-directed mutagenesis, the mutation in a highly conserved region perturbs the structural local environment in POXA1b, leading to a large rearrangement of the enzyme structure. The mutant shows highly reduced activity compared to the wild-type enzyme and is inactive with substrate syringaldazine Pleurotus ostreatus
additional information deletion of the C-terminal extension of 4 or 16 amino acids leads to truncated mutants which lose the high stability at pH 10, while they show an increased stability at pH 5.0, the thermostability is unaltered compared to the wild-type enzyme Pleurotus ostreatus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information construction of enzyme–substrate complexes and molecular dynamics simulations, complex of mutant D205R with 2,5-xylidine, molecular dynamics simulations, overview Pleurotus ostreatus
0.03
-
 syringaldazine pH 6.0, 22°C, recombinant truncated mutant POXA1bDELTA16 Pleurotus ostreatus
0.03
-
 syringaldazine pH 6.0, 22°C, recombinant truncated mutant POXA1bDELTA4 Pleurotus ostreatus
0.04
-
 syringaldazine pH 6.0, 22°C, recombinant wild-type enzyme Pleurotus ostreatus
0.07
-
 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) pH 3.0, 22°C, recombinant wild-type enzyme Pleurotus ostreatus
0.11
-
 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) pH 3.0, 22°C, recombinant truncated mutant POXA1bDELTA16 Pleurotus ostreatus
0.11
-
 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) pH 3.0, 22°C, recombinant truncated mutant POXA1bDELTA4 Pleurotus ostreatus
0.32
-
 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) pH 3.0, 22°C, recombinant truncated mutant POXA1bD205R Pleurotus ostreatus
0.43
-
 2,6-dimethoxyphenol pH 5.3, 22°C, recombinant truncated mutant POXA1bDELTA16 Pleurotus ostreatus
0.43
-
 2,6-dimethoxyphenol pH 5.3, 22°C, recombinant truncated mutant POXA1bDELTA4 Pleurotus ostreatus
0.57
-
 2,6-dimethoxyphenol pH 5.3, 22°C, recombinant wild-type enzyme Pleurotus ostreatus
1.26
-
 2,6-dimethoxyphenol pH 5.3, 22°C, recombinant truncated mutant POXA1bD205R Pleurotus ostreatus

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ laccases contain four copper ions distributed into three sites that, according to their spectroscopic properties, are classified as type 1 Cu, or blue copper centre, type 2 or normal copper centres, and type 3 or coupled binuclear copper centres, overview Pleurotus ostreatus

Organism

Organism UniProt Comment Textmining
Pleurotus ostreatus O60199
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes partially from Saccharomyces cerevisiae strain W303-1A Pleurotus ostreatus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3
-
 partially purified recombinant truncated mutant POXA1bD205R, substrate 2,6-dimethoxyphenol Pleurotus ostreatus
12
-
 partially purified recombinant truncated mutant POXA1bDELTA16, substrate syringaldazine Pleurotus ostreatus
15
-
 partially purified recombinant wild-type enzyme, substrate syringaldazine Pleurotus ostreatus
17
-
 partially purified recombinant truncated mutant POXA1bDELTA4, substrate syringaldazine Pleurotus ostreatus
33
-
 partially purified recombinant truncated mutant POXA1bD205R, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid Pleurotus ostreatus
33
-
 partially purified recombinant truncated mutant POXA1bDELTA16, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid Pleurotus ostreatus
100
-
 partially purified recombinant truncated mutant POXA1bDELTA4, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid Pleurotus ostreatus
129
-
 partially purified recombinant truncated mutant POXA1bDELTA16, substrate 2,6-dimethoxyphenol Pleurotus ostreatus
155
-
 partially purified recombinant wild-type enzyme, substrate 2,6-dimethoxyphenol Pleurotus ostreatus
162
-
 partially purified recombinant wild-type enzyme, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid Pleurotus ostreatus
189
-
 partially purified recombinant truncated mutant POXA1bDELTA4, substrate 2,6-dimethoxyphenol Pleurotus ostreatus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) + O2
-
 Pleurotus ostreatus ?
-
 ?
2,6-dimethoxyphenol + O2
-
 Pleurotus ostreatus ?
-
 ?
syringaldazine + O2
-
 Pleurotus ostreatus ?
-
 ?

Subunits

Subunits Comment Organism
More the POXA1b laccase possesses an unusual C-terminal extension of sixteen amino acids compared to other laccases Pleurotus ostreatus

Synonyms

Synonyms Comment Organism
POXA1b
-
 Pleurotus ostreatus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room tenperature Pleurotus ostreatus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3
-
 assay at, substrate 2,2'-azinobis(3-ethylbenzo-6-thiazolinesulfonic) acid Pleurotus ostreatus
5.3
-
 assay at, substrate 2,6-dimethoxyphenol Pleurotus ostreatus
6
-
 assay at, substrate syringaldazine Pleurotus ostreatus

pH Range

pH Minimum pH Maximum Comment Organism
3 6.4 pH profile with substrate 2,6-dimethoxyphenol Pleurotus ostreatus

pH Stability

pH Stability pH Stability Maximum Comment Organism
3 10 stability of recombinant wild-type and mutant enzymes, overview Pleurotus ostreatus
10
-
 half-life are 30 days Pleurotus ostreatus