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Saccharomyces cerevisiae expressing the native Artemisia annua cytochrome P450 monooxygenase (CYP71AV1) and artemisinic aldehyde D11(13) reductase (DBR2) is used as a whole-cell biocatalyst to produce the immediate artemisinin precursor, dihydroartemisinic acid (DHAA). Introducing artemisinic aldehyde dehydrogenase (ALDH1) from Artemisia annua, which recycles NADPH, results in a significant enhancement in artemisinate titer. Method optimization and evaluation, host screening and cofactor engineering, overview. The co-expression of DBR2 and modulation of abiotic conditions result in a higher yield. Optimal condition are 80 OD yeast cells in 0.05 ml reaction at pH 8.0. Growing the yeast cells at 20°C in galactose medium increases the dihydroartemisinate titer by 50% |
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