title: Enhanced production of coenzyme Q10 by self-regulating the engineered MEP pathway in Rhodobacter sphaeroides.
abstract: Fine-tuning the expression level of an engineered pathway is crucial for the metabolic engineering of a host toward a desired phenotype. However, most engineered hosts suffer from nonfunctional protein expression, metabolic imbalance, cellular burden or toxicity from intermediates when an engineered pathway is first introduced, which can decrease production of the desired product. To circumvent these obstacles, we developed a self-regulation system utilizing the trc/tac promoter, LacI(q) protein and ribosomal binding sites (RBS). With the purpose of improving coenzyme Q10 (CoQ10 ) production by increasing the decaprenyl diphosphate supplement, enzymes DXS, DXR, IDI, and IspD were constitutively overexpressed under the control of the trc promoter in Rhodobacter sphaeroides. Then, a self-regulation system combining a set of RBSs for adjusting the expression of the LacI(q) protein was applied to tune the expression of the four genes, resulting in improved CoQ10 production. Finally, another copy of the tac promoter with the UbiG gene (involved in the ubiquinone pathway of CoQ10 biosynthesis) was introduced into the engineered pathway. By optimizing the expression level of both the upstream and downstream pathway, CoQ10 production in the mutants was improved up to 93.34 mg/L (7.16 mg/g DCW), about twofold of the wild-type (48.25 mg/L, 3.24 mg/g DCW).
authors.affiliation: Department of Chemical and Biological Engineering, Institute of Bioengineering, Zhejiang University, Hangzhou, 310027, PR China.
authors.forename: Wenqiang
authors.initials: W
authors.lastname: Lu
authors.forename: Lidan
authors.initials: L
authors.lastname: Ye
authors.forename: Haoming
authors.initials: H
authors.lastname: Xu
authors.forename: Wenping
authors.initials: W
authors.lastname: Xie
authors.forename: Jiali
authors.initials: J
authors.lastname: Gu
authors.forename: Hongwei
authors.initials: H
authors.lastname: Yu
mesh_terms: D004896:Erythritol / Q000031:analogs & derivatives* / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000235:genetics* / Q000502:physiology; D012242:Rhodobacter sphaeroides / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism
chemical_list: D014451:Ubiquinone; C024989:coenzyme Q10; D004896:Erythritol
keywords: LacIq; coenzyme Q10; constitutive expression; metabolic balance; ribosomal bind site; self-regulation