Cloned (Comment) | Organism |
---|---|
co-expression of C-terminally FLAG-tagged premnaspirodiene oxygenase of Hyoscyamus muticus (HPO) and the Arabidopsis thaliana cytochrome P450 reductase (CPR) in Pichia pastoris strains ADH-C1 and -C3 | Hyoscyamus muticus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | production of the sesquiterpenoid (+)-nootkatone by metabolic engineering of Pichia pastoris by generation of a strain co-expressing the premnaspirodiene oxygenase of Hyoscyamus muticus (HPO) and the Arabidopsis thaliana cytochrome P450 reductase (CPR) that hydroxylates extracellularly added (+)-valencene. Intracellular production of (+)-valencene by co-expression of valencene synthase from Callitropsis nootkatensis resolves the phase-transfer issues of (+)-valencene. Bi-phasic cultivations of Pichia pastoris result in the production of trans-nootkatol, which is oxidized to (+)-nootkatone by an intrinsic Pichia pastoris activity. Additional overexpression of a Pichia pastoris alcohol dehydrogenase and truncated hydroxy-methylglutaryl-CoA reductase (tHmg1p) significantly enhances the (+)-nootkatone yield to 208 mg/l cell culture in bioreactor cultivations. After 12 h of biotransformation about 50% of added (+)-valencene is converted to (+)-nootkatone without residual trans-nootkatol or ot herby-products, but with a moderate overall yield of 48% due to high substrate loss overtime. HPO,CPR and ADH-C3 protein levels are only marginally decreased by co-overexpression of tHMG1 | Hyoscyamus muticus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-valencene + [reduced NADPH-hemoprotein reductase] + O2 | Hyoscyamus muticus | enzyme HPO-mediated biohydroxylation of (+)-valencene | alpha-nootkatol + beta-nootkatol + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
additional information | Hyoscyamus muticus | co-expression of enzyme HPO and cytochrome P450 reductase (CPR) leads to the hydroxylation of externally added (+)-valencene to trans-nootkatol in Pichia pastoris cells | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Hyoscyamus muticus | A6YIH8 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-valencene + [reduced NADPH-hemoprotein reductase] + O2 | enzyme HPO-mediated biohydroxylation of (+)-valencene | Hyoscyamus muticus | alpha-nootkatol + beta-nootkatol + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
additional information | co-expression of enzyme HPO and cytochrome P450 reductase (CPR) leads to the hydroxylation of externally added (+)-valencene to trans-nootkatol in Pichia pastoris cells | Hyoscyamus muticus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CYP71D55 | gene name, UniProt | Hyoscyamus muticus |
HPO | - |
Hyoscyamus muticus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH-hemoprotein reductase | A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases, EC 1.14.14._ | Hyoscyamus muticus |