Metabolic Flow Analysis

Our solution: VersaTobacco

VersaTobacco can be mainly divided into two parts: the construction of the low-chlorogenic acid content Nicotiana benthamiana chassis and functional verification of the chassis.

The chlorogenic acid synthesis pathway of tobacco mainly involves six key enzymes: phenylalnine ammonialyase(PAL)、cinnamic acid 4-hydroxylase(C4H), 4-coumarate-CoA ligase(4CL)、p-cose(HCT) and hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase(HQT)[5-8]

We used the COBRApy toolkit to perform flux balance anSMM) of Nicotiana benthamiana and found that knocking out the HQT down-regulated the chlorogenic acid production response. Significantly correlated with the upregulation of other metabolic flux reactions.

Part I: Chassis Construction

Constructing NbHQTs knockout N.benthamiana by CRISPR/Cas9

The CRISPR/C heterolo-Cas9 technology. This approach aims to shiftic biology and increasing the yield of medicinal secondary metabolites.

Synthesis of resveratrol

Hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase,HQT catalyzes the conversion of p-coumaroyl-CoA to produce p-coumic pathways to enhance the production of downstream products.

Resveratrol (3,5,4'-trihydroxystilbene) presents in more than 70 species of plants, particulaely in grapes. It is mostly presifferent tilbene synthom Vitis vi synthase [16], trying to synthesize resveratrol in tobacco.

In addition, modificationsh can reduce the toxic effects of resveratrol on plants [14] and further increase the production of resveratrol.