EC Number   |
Protein Variants |
Reference |
|---|
   1.14.14.151 | A484I |
the mutation results in a catalytically compromised enzyme |
712360 |
| 1.14.14.151 | more |
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 |
745687 |
| 1.14.14.151 | V366S |
the mutation dramatically reduces overall enzyme activity |
712360 |
| 1.14.14.151 | V480I/A484I |
mutant with improved catalytic efficiency |
712360 |
| 1.14.14.151 | V480S |
the mutation improves the kcat for the conversion of (-)-vetispiradiene to solavetivol about 2fold |
712360 |
| 1.14.14.151 | V482I |
mutant with improved catalytic efficiency |
712360 |
| 1.14.14.151 | V482I |
the mutation improves the kcat for the conversion of (-)-vetispiradiene to solavetivol about 2fold |
712360 |
| 1.14.14.151 | V482I/A484I |
the mutant possesses a 5fold improvement in its catalytic efficiency for nootkatol biosynthesis and a 10fold improvement for 2beta-hydroxy-epiaristolochene biosynthesis |
712360 |
