EC Number | Cloned (Comment) | Organism |
---|---|---|
1.13.11.B6 | recombinant expression in Pichia pastoris | Nakataea oryzae |
1.13.11.45 | recombinant expression of wild-type and mutant enzymes in Pichia pastoris | Gaeumannomyces graminis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.13.11.B6 | L350M | site-directed mutagenesis, the mutant shows increased specificity for 13S-lipoxygenation, EC 1.13.11.12, altered substrate specificity/regiospecific activity, overview | Nakataea oryzae |
1.13.11.B6 | additional information | replacement Leu350 in 9S-MnLOX and changes the regiospecific oxidation of 18:2 n-6 in a consistent way, but the n-3 double bond of 18:3 n-3 can reduce this effect | Nakataea oryzae |
1.13.11.45 | F347A | site-directed mutagenesis, the mutant oxidizes octadeca-9,11-dienoic acid to 11-hydroperoxyoctadecadienoate/(13R)-hydroperoxyoctadecadienoate/(9S)-hydroperoxyoctadecadienoate in an initial ratio of 50/42/8 and with almost complete consumption of 11-hydroperoxyoctadecadienoate to a ratio of 2/84/14. The (9E,11Z,15E)-octadeca-9,11,15-trienoic acid is transformed to 11- and (13R)-hydroperoxyoctadecatrienoic acid and to traces of (9S)-hydroperoxyoctadecatrienoic acid, essentially as native 13R-MnLOX | Gaeumannomyces graminis |
1.13.11.45 | F347L | site-directed mutagenesis, (9E,11Z,15E)-octadeca-9,11,15-trienoic acid is oxidized to a 2:3 mixture of (9S)- and (13R)-dihydroxyoctadecatrienoic acid | Gaeumannomyces graminis |
1.13.11.45 | F347V | site-directed mutagenesis | Gaeumannomyces graminis |
1.13.11.45 | additional information | replacement Phe347 in 13R-MnLOX and changes the regiospecific oxidation of 18:2 n-6 in a consistent way, but the n-3 double bond of 18:3n-3 can reduce this effect. Mutations are designed to convert the pentamer motif to a hexamer motif to mimic FeLOX, but the mutants with the His-Val-Leu-Phe-Thr-His and His-Val-Leu-Phe-Gly-His motives are inactive, overview | Gaeumannomyces graminis |
EC Number | General Stability | Organism |
---|---|---|
1.13.11.B6 | 9S-LOX activity is less robust during expression and purification, possibly due to proteolysis, stability is not improved by addition of 0.001 mM pepstatin A, 0.001 mM leupeptin, or 1 mM EDTA to the growth medium | Nakataea oryzae |
1.13.11.45 | 9S-LOX activity is less robust during expression and purification, possibly due to proteolysis, stability is not improved by addition of 0.001 mM pepstatin A, 0.001 mM leupeptin, or 1 mM EDTA to the growth medium | Gaeumannomyces graminis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.B6 | 0.0037 | - |
(9Z,12Z)-octadeca-9,12-dienoic acid | pH 9.0, 22°C | Nakataea oryzae |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
1.13.11.B6 | extracellular | the enzyme is secreted | Nakataea oryzae | - |
- |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.13.11.B6 | Fe2+ | Mn:Fe ratio is 1:0.05 | Nakataea oryzae | |
1.13.11.B6 | Mn2+ | manganese 9S-lipoxygenase, 9S-LOX contains catalytic manganese, Mn:protein ratio is about 0.2:1, while the Mn:Fe ratio is 1:0.05 | Nakataea oryzae | |
1.13.11.B6 | additional information | the catalytic metal of the 9S-LOX is manganese and not iron | Nakataea oryzae | |
1.13.11.45 | Mn2+ | manganese 13R-lipoxygenase contains catalytic manganese, the catalytic domain of 13R-MnLOX contains a pentamer motif flanked by two His metal ligands, His-Val-Leu-Phe-His, in the presumed manganese binding region | Gaeumannomyces graminis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.13.11.B6 | 66000 | - |
x * 80000-90000, recombinant glycosylated enzyme, SDS-PAGE, x * 66000, recombinant deglycosylated enzyme, SDS-PAGE | Nakataea oryzae |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.13.11.B6 | Nakataea oryzae | - |
- |
- |
1.13.11.B6 | Nakataea oryzae CBS 288.54 | - |
- |
- |
1.13.11.45 | Gaeumannomyces graminis | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.13.11.B6 | recombinant enzyme from Pichia pastoris by hydrophobic interaction chromatography, dialfiltration, and gel filtration | Nakataea oryzae |
1.13.11.45 | recombinant wild-type and mutant enzymes from Pichia pastoris by hydrophobic interaction chromatography, dialfiltration, and gel filtration | Gaeumannomyces graminis |
EC Number | Source Tissue | Comment | Organism | Textmining |
---|---|---|---|---|
1.13.11.B6 | mycelium | - |
Nakataea oryzae | - |
EC Number | Storage Stability | Organism |
---|---|---|
1.13.11.B6 | purified recombinant 9S-MnLOX can be stored for several weeks at 4°C with retained activity | Nakataea oryzae |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.B6 | (9E,11Z,15E)-octadeca-9,11,15-trienoic acid + O2 | - |
Nakataea oryzae | ? | - |
? | |
1.13.11.B6 | (9E,11Z,15E)-octadeca-9,11,15-trienoic acid + O2 | - |
Nakataea oryzae CBS 288.54 | ? | - |
? | |
1.13.11.B6 | (9Z,12Z)-octadeca-9,12-dienoic acid + O2 | alpha-linoleate is converted via two intermediates, (9Z,11S,12Z)-11-hydroperoxy-9,12-octadecenoate and (9Z,11E,13R)-13-hydroperoxy-9,12-octadecadienoate, which are isomerized to the end product, probably after oxidation to peroxyl radicals, beta-fragmentation, and oxygen insertion at C-9 | Nakataea oryzae | (9S,10E,12Z)-9-hydroperoxy-octadeca-10,12-dienoate | - |
? | |
1.13.11.B6 | (9Z,12Z)-octadeca-9,12-dienoic acid + O2 | alpha-linoleate is converted via two intermediates, (9Z,11S,12Z)-11-hydroperoxy-9,12-octadecenoate and (9Z,11E,13R)-13-hydroperoxy-9,12-octadecadienoate, which are isomerized to the end product, probably after oxidation to peroxyl radicals, beta-fragmentation, and oxygen insertion at C-9 | Nakataea oryzae CBS 288.54 | (9S,10E,12Z)-9-hydroperoxy-octadeca-10,12-dienoate | - |
? | |
1.13.11.B6 | (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2 | gamma-linoleate is oxidized at C-9, C-11, and C-13 | Nakataea oryzae | ? | - |
? | |
1.13.11.B6 | (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2 | gamma-linoleate is oxidized at C-9, C-11, and C-13 | Nakataea oryzae CBS 288.54 | ? | - |
? | |
1.13.11.B6 | additional information | cf. EC 1.13.11.45, LC- and GC-MS analysis product analysis, overview. Authentic (11S)-hydroperoxyoctadecadienoate is rapidly transformed to 9-hydroperoxyoctadecadienoate by 9S-MnLOX, and only traces of 13-hydroperoxyoctadecadienoate. 9S-MnLOX also partly transforms authentic (13R)-hydroperoxyoctadecadienoate to (9S)-hydroperoxyoctadecadienoate | Nakataea oryzae | ? | - |
? | |
1.13.11.B6 | additional information | cf. EC 1.13.11.45, LC- and GC-MS analysis product analysis, overview. Authentic (11S)-hydroperoxyoctadecadienoate is rapidly transformed to 9-hydroperoxyoctadecadienoate by 9S-MnLOX, and only traces of 13-hydroperoxyoctadecadienoate. 9S-MnLOX also partly transforms authentic (13R)-hydroperoxyoctadecadienoate to (9S)-hydroperoxyoctadecadienoate | Nakataea oryzae CBS 288.54 | ? | - |
? | |
1.13.11.45 | (9Z,12Z)-octadeca-9,12-dienoic acid + O2 | alpha-linoleate is converted via two intermediates, (11S)-hydroperoxy-(9Z,12Z)-octadecenoate and (13R)-hydroperoxy-(9Z,11E)-octadecadienoate, which are isomerized to the end product, probably after oxidation to peroxyl radicals, beta-fragmentation, and oxygen insertion at C-9 | Gaeumannomyces graminis | (9S)-hydroperoxy-octadeca-10,12-dienoate | - |
? | |
1.13.11.45 | (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid + O2 | gamma-linoleate is oxidized at C-9, C-11, and C-13 | Gaeumannomyces graminis | (10E,12E,14E)-9,16-dihydroxy-octadeca-10,12,14-trienoate | - |
? | |
1.13.11.45 | additional information | cf. EC 1.13.11.58, LC- and GC-MS analysis product analysis, overview | Gaeumannomyces graminis | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.13.11.B6 | ? | x * 80000-90000, recombinant glycosylated enzyme, SDS-PAGE, x * 66000, recombinant deglycosylated enzyme, SDS-PAGE | Nakataea oryzae |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.13.11.B6 | linoleate 9 S-lipoxygenase | - |
Nakataea oryzae |
1.13.11.B6 | manganese 9S-lipoxygenase | - |
Nakataea oryzae |
1.13.11.B6 | S-LOX | - |
Nakataea oryzae |
1.13.11.45 | 13 R -MnLOX | - |
Gaeumannomyces graminis |
1.13.11.45 | manganese lipoxygenase | - |
Gaeumannomyces graminis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.13.11.B6 | 22 | - |
assay at | Nakataea oryzae |
1.13.11.45 | 22 | - |
assay at | Gaeumannomyces graminis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.B6 | 8.3 | - |
(9Z,12Z)-octadeca-9,12-dienoic acid | pH 9.0, 22°C | Nakataea oryzae |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.13.11.B6 | 9 | - |
assay at | Nakataea oryzae |
1.13.11.45 | 9 | - |
assay at | Gaeumannomyces graminis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.13.11.B6 | evolution | 9S-LOX contains catalytic manganese, and its sequence can be aligned with 77% identity to 13R-LOX with catalytic manganese lipoxygenase of the Take-all fungus. Alterations in the Sloane determinant of 9S-LOX and 13R-MnLOX with larger and smaller hydrophobic residues interconvert the regiospecifi c oxidation of 18:2 n-6, presumably by altering the substrate position in relation to oxygen insertion | Nakataea oryzae |
1.13.11.B6 | additional information | 9S-MnLOX catalyzes hydrogen abstraction at C-11 and oxygenation at C-9 and C-11 in a suprafacial manner in analogy with 13R-MnLOX, EC 1.13.11.45 | Nakataea oryzae |
1.13.11.45 | evolution | 9S-LOX contains catalytic manganese, and its sequence can be aligned with 77% identity to 13R-LOX with catalytic manganese lipoxygenase of the Take-all fungus. Alterations in the Sloane determinant of 9S-LOX and 13R-MnLOX with larger and smaller hydrophobic residues interconvert the regiospecific oxidation of 18:2n-6, presumably by altering the substrate position in relation to oxygen insertion | Gaeumannomyces graminis |
1.13.11.45 | additional information | alterations in the Sloane determinant of 9S-LOX and 13R-MnLOX with larger and smaller hydrophobic residues interconvert the regiospecific oxidation of 18:2n-6, presumably by altering the substrate position in relation to oxygen insertion. The catalytic domain of 13R-MnLOX contains a pentamer motif flanked by two His metal ligands, His-Val-Leu-Phe-His, in the presumed manganese binding region. 9S-MnLOX, EC 1.13.11.58, catalyzes hydrogen abstraction at C-11 and oxygenation at C-9 and C-11 in a suprafacial manner in analogy with 13R-MnLOX | Gaeumannomyces graminis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.B6 | 0.45 | - |
(9Z,12Z)-octadeca-9,12-dienoic acid | pH 9.0, 22°C | Nakataea oryzae |