Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in Pichia pastoris | Gaeumannomyces graminis |
Protein Variants | Comment | Organism |
---|---|---|
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 |
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 |
F347V | site-directed mutagenesis | Gaeumannomyces graminis |
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 |
General Stability | Organism |
---|---|
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 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
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 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Gaeumannomyces graminis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Pichia pastoris by hydrophobic interaction chromatography, dialfiltration, and gel filtration | Gaeumannomyces graminis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(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 | - |
? | |
(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 | - |
? | |
additional information | cf. EC 1.13.11.58, LC- and GC-MS analysis product analysis, overview | Gaeumannomyces graminis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
13 R -MnLOX | - |
Gaeumannomyces graminis |
manganese lipoxygenase | - |
Gaeumannomyces graminis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Gaeumannomyces graminis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
assay at | Gaeumannomyces graminis |
General Information | Comment | Organism |
---|---|---|
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 |
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 |