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Literature summary for 1.14.18.1 extracted from

  • Pretzler, M.; Bijelic, A.; Rompel, A.
    Heterologous expression and characterization of functional mushroom tyrosinase (AbPPO4) (2017), Sci. Rep., 7, 1810 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
additional information enzymatic activity may be induced in the reaction mixture by the addition of a ionic detergent, e.g. SDS Agaricus bisporus
SDS activates at 2 mM Agaricus bisporus

Cloned(Commentary)

Cloned (Comment) Organism
Agaricus bisporus possesses genes coding for six different tyrosinases (AbPPO1-AbPPO6), full-length gene AbPPO4 (1-565) DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally GST-tagged tyrosinase 4 from gene AbPPO4 containing 23 mutations in Escherichia coli as latent enzyme Agaricus bisporus

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant detagged latent tyrosinase, X-ray diffraction structure determination and analysis. In contrast to the crystals obtained with the enzyme isolated from the natural source which contains two different chains in the asymmetric unit (one latent and one active protein) and does only form in the presence of sodium hexatungstotellurate(VI) (Na6[TeW6O24] x 22 H2O), the recombinant enzyme forms crystals containing exclusively the latent tyrosinase. The latent chain of AbPPO4 isolated from Agaricus bisporu, PDB ID 4OUA, chain B, is aligned with the heterologously produced protein, PDB ID 5M6B, chain B Agaricus bisporus

Protein Variants

Protein Variants Comment Organism
additional information mutations relative to GQ354802 mRNA for the reference sequence for AbPPO4, Uniprot ID C7FF05 : C21T, T168C, T306C, T362C (V121A), T483C, A504C, G536A (S179N), A540C, T717C, T735C, G1089A, C1104T, C1131G, G1218A, C1359T, T1449C, C1458T, A1521G, C1650T, T1686C, T1704C, G1717A (V573I), G1783A (A595T) in the wild-type AbPPO4 and C21T, G97A (V33I), G133T (A45S), T168C, G301A (V101I), G324C, T362C (V121A), T483C, A504C, G536A (S179N), A540C, G563A (R188K), C618T, C620G (A207G), T171C, T735C, G1089A, C1131G, T1172A & C1173A (L391Q), G1783A (A595T) in the mutant AbPPO4DELTA(A436-A580). Construction of a truncated enzyme mutant encoding only the main domain of the tyrosinase up to residue S383, which does not exhibit any tyrosinase activity Agaricus bisporus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.14
-
L-tyrosine pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
4.65
-
catechol pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
9.53
-
tyramine pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
12.5
-
phenol pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
15.2
-
dopamine pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
26.1
-
L-Dopa pH 6.8, 25°C, recombinant enzyme Agaricus bisporus

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ a type III copper-containing metalloenzyme Agaricus bisporus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 L-dopa + O2 Agaricus bisporus
-
2 dopaquinone + 2 H2O
-
?
L-tyrosine + O2 Agaricus bisporus
-
dopaquinone + H2O
-
?

Organism

Organism UniProt Comment Textmining
Agaricus bisporus C7FF05 mushroom tyrosinase describes a mixture of isoenzymes containing a number of enzymatic side-activities
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification the latent recombinant tyrosinase 4 enzyme can be activated by limited proteolysis with proteinase K, which cleaves the polypeptide chain after K382, only one The latent enzyme can amino acid before the main in-vivo activation site. Activation by proteinase K is more efficient compared to activation by trypsin Agaricus bisporus

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, followed by tag cleavage through specific protease HRV 3 C Agaricus bisporus

Source Tissue

Source Tissue Comment Organism Textmining
commercial preparation mushroom tyrosinase Agaricus bisporus
-
fruiting body tyrosinase 4 Agaricus bisporus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.22
-
purified recombinant enzyme, substrate L-tyrosine, pH 6.8, 25°C Agaricus bisporus

Storage Stability

Storage Stability Organism
4°C, purified recombinant detagged enzyme, after 1 year of storage in 10 mM HEPES, pH 7.5, the enzyme is still active Agaricus bisporus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 L-dopa + O2
-
Agaricus bisporus 2 dopaquinone + 2 H2O
-
?
catechol + O2 reaction of EC 1.10.3.1 Agaricus bisporus ?
-
?
dopamine + O2
-
Agaricus bisporus ?
-
?
L-tyrosine + O2
-
Agaricus bisporus dopaquinone + H2O
-
?
additional information the proteolytically activated mushroom tyrosinase shows over 50% of its maximal activity in the range of pH 5-10 and accepts a wide range of substrates including mono- and diphenols, flavonols and chalcones. The activated AbPPO4 catalyzes both reactions observed for tyrosinase. The catechol oxidase activity proceeds typically with a rate two orders of magnitude faster than the hydroxylation and oxidation of monophenols. Of the tested substrates the enzyme exhibits the highest affinity and the lowest reaction rate for L-tyrosine. Activated AbPPO4 discriminates between enantiomers of tyrosine showing pronounced differences in the rate of the tyrosinase reaction. For tyrosine 1 mM of the L-enantiomer is converted at a rate of 1.22 U/mg, which is 2.58times faster than the rate on D-tyrosine. A slight increase in enantioselectivity is seen for the methyl ester of tyrosine Agaricus bisporus ?
-
?
phenol + O2
-
Agaricus bisporus ?
-
?
tyramine + O2
-
Agaricus bisporus ?
-
?

Subunits

Subunits Comment Organism
? x * 52000, recombinant wild-type enzyme, SDS-PAGE Agaricus bisporus

Synonyms

Synonyms Comment Organism
AbPPO4
-
Agaricus bisporus
mushroom tyrosinase
-
Agaricus bisporus
polyphenol oxidase 4 UniProt Agaricus bisporus
tyrosinase 4
-
Agaricus bisporus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Agaricus bisporus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.77
-
L-tyrosine pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
11.3
-
phenol pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
15.6
-
tyramine pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
1080
-
catechol pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
1810
-
L-Dopa pH 6.8, 25°C, recombinant enzyme Agaricus bisporus
2540
-
dopamine pH 6.8, 25°C, recombinant enzyme Agaricus bisporus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.8
-
assay at Agaricus bisporus

pH Range

pH Minimum pH Maximum Comment Organism
5 10 the proteolytically activated mushroom tyrosinase shows over 50% of its maximal activity in the range of pH 5-10 and accepts a wide range of substrates including mono- and diphenols, flavonols and chalcones Agaricus bisporus

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 10 in the pH-range from pH 5-10 the enzyme retains more than 50% of its activity Agaricus bisporus

General Information

General Information Comment Organism
physiological function tyrosinases are an ubiquitous group of copper-containing metalloenzymes that hydroxylate and oxidize phenolic molecules. Tyrosinase catalyzes the o-hydroxylation of monophenols (EC 1.14.18.1) and the oxidation of o-diphenols (cf. EC 1.10.3.1) Agaricus bisporus