Combining in silico and in vitro approaches to understand the involvement of MEP and shikimate pathway in Agrobacterium tumefaciens for enhanced CoQ10 production.

Abstract

To carry out integrated comparative metabolic pathway analysis to understand Coenzyme Q10 (CoQ10) production in Agrobacterium tumefaciens. Comparative analysis of the CoQ10 metabolic pathways in 10 organisms using a G2KO program and KEGG database shed light on the completeness of the production pathway in A. tumefaciens. The specific role of the key precursors and the enzymes in the metabolic network was further validated using pathway inhibitors and enhancers. While the use of fosmidomycin and glyphosate displayed 54.54%-99% suppression in CoQ10 production, supplementation of polyprenyl pyrophosphate from the MEP (methylerythritol 4-phosphate) pathway and 4-hydroxybenzoate precursor from the shikimate pathway, enhanced CoQ10 production by 2.3-fold. The present study provides a comprehensive understanding of the CoQ10 biosynthetic pathway in A. tumefaciens, which would assist rational metabolic engineering strategies for boosting CoQ10 biosynthesis.