Cloned (Comment) | Organism |
---|---|
- |
Penicillium roqueforti |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the replacement of Trp 334 with para-substituted phenylalanines of increasing electron-withdrawing properties leads to a progressive accumulation of germacrene A that shows a good correlation with the interaction energies of simple cations such as Na+ with substituted benzenes. Evidence for the stabilizing role played by Trp334 in aristolochene synthase catalysis for the energetically demanding transformation of germacrene A to eudesmane cation. Replacement of tryptophan by para-substituted phenylalanines with strong electron-withdrawing substituents has only minor effects on the KM values of the reaction but leads to approximately 30fold decreases of kcat relative to mutant W334F. Noncanonical substitutions lead to the following ratios of products: W334naphthyl 78% aristolochene, 22% germacrene A, W334-p-chlorophenylalanine or W334-p-fluorophenylalanine 57% aristolochene, 43% germacrene A, W334-p-trifluoromethylphenylalanine 31% aristolochene, 69% germacrene A, W334-p-nitrophenylalanine 23% aristolochene, 77% germacrene A | Penicillium roqueforti |
W334F | ratio of products: 82% aristolochene, 18% germacrene A | Penicillium roqueforti |
W334H | ratio of products: 10% aristolochene, 90% germacrene A | Penicillium roqueforti |
W334L | ratio of products: 2% aristolochene, 98% germacrene A | Penicillium roqueforti |
W334Y | ratio of products: 62% aristolochene, 38% germacrene A | Penicillium roqueforti |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00013 | - |
(2E,6E)-farnesyl diphosphate | mutant W334H, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.00028 | - |
(2E,6E)-farnesyl diphosphate | mutant W334F, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.00053 | - |
(2E,6E)-farnesyl diphosphate | wild-type, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.00075 | - |
(2E,6E)-farnesyl diphosphate | mutant W334L, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.001 | - |
(2E,6E)-farnesyl diphosphate | mutant W334Y, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Penicillium roqueforti | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2E,6E)-farnesyl diphosphate | reaction proceeds via germacrene A and eudesmane cation. The amount of germacrene A generated by mutant enzymes serves as a measure of the stabilization of eudesmane cation | Penicillium roqueforti | aristolochene + diphosphate | wild-type, about 8% of germacrene A as by-product. His-tagged wild-type, about 4% of germacrene A | ? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0003 | - |
(2E,6E)-farnesyl diphosphate | mutant W334H, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.0027 | - |
(2E,6E)-farnesyl diphosphate | mutant W334L, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.006 | - |
(2E,6E)-farnesyl diphosphate | mutant W334F, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.007 | - |
(2E,6E)-farnesyl diphosphate | mutant W334Y, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
0.084 | - |
(2E,6E)-farnesyl diphosphate | wild-type, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.56 | - |
(2E,6E)-farnesyl diphosphate | mutant W334H, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
3.56 | - |
(2E,6E)-farnesyl diphosphate | mutant W334L, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
6.95 | - |
(2E,6E)-farnesyl diphosphate | mutant W334Y, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
22.6 | - |
(2E,6E)-farnesyl diphosphate | mutant W334F, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti | |
158.5 | - |
(2E,6E)-farnesyl diphosphate | wild-type, pH not specified in the publication, temperature not specified in the publication | Penicillium roqueforti |