초록
<P>Identification of diterpene synthase-encoding genes together with synthetic biology technology offers an opportunity for the biosynthesis of <I>cis</I>-abienol. The methylerythritol phosphate (MEP) and the mevalonate (MVA) pathways were both engineered for <I>cis</I>-abienol production in <I>Escherichia coli</I>, which improved the <I>cis</I>-abienol yield by approximately 7-fold and 31-fold, respectively, compared to the yield obtained by overexpression of the MEP pathway alone or the original MEP pathway. Furthermore, systematic optimization of <I>cis</I>-abienol biosynthesis was performed, such as diterpene synthase screening and two-phase cultivation. The combination of bifunctional class I/II <I>cis</I>-abienol synthase from <I>Abies balsamea</I> (<I>Ab</I>CAS) and class II abienol synthase from <I>Salvia sclarea</I> (<I>Ss</I>TPS2) was found to be the most effective. By using isopropyl myristate as a solvent in two-phase cultivation, <I>cis</I>-abienol production reached 634.7 mg/L in a fed-batch bioreactor. This work shows the possibility of <I>E. coli</I> utilizing glucose as a carbon source for <I>cis</I>-abienol biosynthesis through a modified pathway.</P><BR>[FIG OMISSION]</BR>