Coupling limonene formation and oxyfunctionalization by mixed-culture resting cell fermentation

Biotechnology and Bioengineering
Christian WillrodtMattijs K Julsing

Abstract

Metabolic engineering strategies mark a milestone for the fermentative production of bulk and fine chemicals. Yet, toxic products and volatile reaction intermediates with low solubilities remain challenging. Prominent examples are artificial multistep pathways like the production of perillyl acetate (POHAc) from glucose via limonene. For POHAc, these limitations can be overcome by mixed-culture fermentations. A limonene biosynthesis pathway and cytochrome P450 153A6 (CYP153A6) as regioselective hydroxylase are used in two distinct recombinant E. coli. POHAc formation from glucose in one recombinant cell was hindered by ineffective coupling of limonene synthesis and low rates of oxyfunctionalization. The optimization of P450 gene expression led to the formation of 6.20 ± 0.06 mg gcdw (-1) POHAc in a biphasic batch cultivation with glucose as sole carbon and energy source. Increasing the spatial proximity between limonene synthase and CYP153A6 by a genetic fusion of both enzymes changed the molar limonene/POHAc ratio from 3.2 to 1.6. Spatial separation of limonene biosynthesis from its oxyfunctionalization improved POHAc concentration 3.3-fold to 21.7 mg L(-1) as compared to a biphasic fermentation. Mixed-cultures of E. coli BL21...Continue Reading

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Citations

Apr 4, 2015·Current Opinion in Biotechnology·Christian WillrodtMattijs K Julsing
Jun 1, 2016·Metabolic Engineering·Haoran Zhang, Xiaonan Wang
Nov 18, 2015·Biotechnology and Bioengineering·Christian WillrodtMattijs K Julsing
Jun 8, 2017·MBio·J Andrew JonesMattheos A G Koffas
Apr 25, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jascha RolfStephan Lütz
Jul 19, 2017·Biotechnology Journal·Marvin KadischAndreas Schmid
Dec 29, 2020·Microbial Biotechnology·Lisa BretschneiderRohan Karande
Dec 22, 2019·Biotechnology Advances·Xiujuan QianKatrin Ochsenreither
Sep 4, 2020·Biotechnology Advances·Yuyao RenYongjin J Zhou

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