Cloned (Comment) | Organism |
---|---|
gene Mco1, detailed sequence comparisons and phylogenetic analysis, recombinant expression of N-terminally His-tagged wild-type and truncated mutant enzymes in Escherichia coli | Acidomyces acidophilus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a truncated enzyme mutant by deletion of the first 83 amino acids containing the putative N-terminal transmembrane helix in order to obtain a soluble protein. Expression of the DELTA-N-truncated protein leads to the production of an insoluble protein that presents a molecular weight estimated at 75 kDa. The truncated protein loses the N-terminal His-tag during or after renaturation step but retains to keep the expected molecular weight, the truncated enzyme shows slightly reduced activity compared to wild-type | Acidomyces acidophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
Fe(II) | recombinant wild-type enzyme, pH 5.0, 25°C | Acidomyces acidophilus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | no predicted signal peptide, but a putative N-terminal transmembrane helix, from amino acid 61-83, is predicted from the enzyme sequence. AaMco1 presents a predicted N-terminal transmembrane helix and lacks the COOH-terminal transmembrane domain common to ferroxidases | Acidomyces acidophilus | 16020 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | a multicopper oxidase, dependent on copper for catalytic activity, sequence motifs for copper binding sites, overview | Acidomyces acidophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4 Fe(II) + 4 H+ + O2 | Acidomyces acidophilus | - |
4 Fe(III) + 2 H2O | - |
? | |
4 Fe(II) + 4 H+ + O2 | Acidomyces acidophilus 26774 | - |
4 Fe(III) + 2 H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acidomyces acidophilus | M9PLY0 | - |
- |
Acidomyces acidophilus 26774 | M9PLY0 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His-tagged wild-type and truncated mutant enzymes from Escherichia coli by immobilized metal affinity chromatography first under denaturing conditions and second as renaturation step by metal affinity chromatography under native conditions | Acidomyces acidophilus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.69 | - |
purified recombinant N-terminally truncated enzyme mutant, pH 5.0, 25°C | Acidomyces acidophilus |
0.702 | - |
purified recombinant wild-type enzyme, pH 5.0, 25°C | Acidomyces acidophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4 Fe(II) + 4 H+ + O2 | - |
Acidomyces acidophilus | 4 Fe(III) + 2 H2O | - |
? | |
4 Fe(II) + 4 H+ + O2 | - |
Acidomyces acidophilus 26774 | 4 Fe(III) + 2 H2O | - |
? | |
additional information | AaMco1 has ferroxidase activity. AaMco1 is also able to oxidize the N,N-dimethyl-pphenylenediamine dihydrochloride (DMPPDA) compound. Ferroxidase activity of the purified protein is measured by the ferrozine assay. No activity with 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS), 2,6-dimethoxyphenol, and 2-L-ascorbic acid | Acidomyces acidophilus | ? | - |
? | |
additional information | AaMco1 has ferroxidase activity. AaMco1 is also able to oxidize the N,N-dimethyl-pphenylenediamine dihydrochloride (DMPPDA) compound. Ferroxidase activity of the purified protein is measured by the ferrozine assay. No activity with 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS), 2,6-dimethoxyphenol, and 2-L-ascorbic acid | Acidomyces acidophilus 26774 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | AaMco1 presents a predicted N-terminal transmembrane helix and lacks the COOH-terminal transmembrane domain common to ferroxidases, structure comparisons, overview | Acidomyces acidophilus |
Synonyms | Comment | Organism |
---|---|---|
MCO1 | - |
Acidomyces acidophilus |
multicopper oxidase 1 | - |
Acidomyces acidophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Acidomyces acidophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
assay at | Acidomyces acidophilus |
General Information | Comment | Organism |
---|---|---|
evolution | in addition to the classical classification, another cluster containing AaMco1 and filamentous ascomycete hypothetical proteins, putative multicopper oxidases (MCOs) or proteins called ascorbate oxidases is identified on the basis of sequence similarity. This group is named ascomycete MCOs. Neighbor joining tree of multicopper oxidase amino acid sequences, phylogenetic analysis, overview | Acidomyces acidophilus |