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to increase the low metabolic flux through PPDS and increase the productivity of protopanaxadiol production, enzyme PPDS is modified through transmembrane domain truncation and construction of self-sufficient PPDS-ATR1 fusion proteins. The fusion enzymes exhibit approximately 4.5fold increase in catalytic activity and 71.1% increase in protopanaxadiol production compared with PPDS and ATR1 co-expression. The engineered yeast carrying fusion protein effectively converts 96.8% of dammarenediol-II into protopanaxadiol, bioreactor in fed-batch fermentation. Construction of four fusion proteins named as PPDS-linker1-46tATR1, PPDSlinker2-46tATR1, PPDS-linker3-46tATR1, and PPDS-nolinker-46tATR1, respectively. In addition, 31tPPDS heme domain and 46tATR1 reductase domain are linked by polypeptide GSTSSGSG. Method optimization, overview. Evaluation of the oxidative stress in yeast cells induced by co-expression of PPDS and ATR1 |
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