title: Carbon partitioning to the terpenoid biosynthetic pathway enables heterologous β-phellandrene production in Escherichia coli cultures.
abstract: Escherichia coli was used as a microbial system for the heterologous synthesis of β-phellandrene, a monoterpene of plant origin with several potential commercial applications. Expression of Lavandula angustifolia β-phellandrene synthase (PHLS), alone or in combination with Picea abies geranyl-diphosphate synthase in E. coli, resulted in no β-phellandrene accumulation, in sharp contrast to observations with PHLS-transformed cyanobacteria. Lack of β-phellandrene biosynthesis in E. coli was attributed to the limited endogenous carbon partitioning through the native 2-C-methylerythritol-4-phosphate (MEP) pathway. Heterologous co-expression of the mevalonic acid pathway, enhancing cellular carbon partitioning and flux toward the universal isoprenoid precursors, isopentenyl-diphosphate and dimethylallyl-diphosphate, was required to confer β-phellandrene production. Differences in endogenous carbon flux toward the synthesis of isoprenoids between photosynthetic (Synechocystis) and non-photosynthetic bacteria (E. coli) are discussed in terms of differences in the regulation of carbon partitioning through the MEP biosynthetic pathway in the two systems.
authors.affiliation: Department of Plant and Microbial Biology, University of California, 111 Koshland Hall, Berkeley, CA, 94720-3102, USA.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Formighieri
authors.forename: Anastasios
authors.initials: A
authors.lastname: Melis
mesh_terms: D053898:Biosynthetic Pathways; D002244:Carbon / Q000378:metabolism; D000081005:Cyclohexane Monoterpenes; D053138:Cyclohexenes / Q000378:metabolism*; D004896:Erythritol / Q000031:analogs & derivatives / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D045782:Hemiterpenes / Q000378:metabolism; D027523:Lavandula / Q000201:enzymology / Q000235:genetics; D008798:Mevalonic Acid / Q000378:metabolism; D039821:Monoterpenes / Q000378:metabolism*; D009943:Organophosphorus Compounds / Q000378:metabolism; D028222:Picea / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D013403:Sugar Phosphates / Q000378:metabolism; D046939:Synechocystis / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism; D014169:Transformation, Bacterial
chemical_list: C114232:2-C-methylerythritol 4-phosphate; D000081005:Cyclohexane Monoterpenes; D053138:Cyclohexenes; D029968:Escherichia coli Proteins; D045782:Hemiterpenes; D039821:Monoterpenes; D009943:Organophosphorus Compounds; D011994:Recombinant Proteins; D013403:Sugar Phosphates; D013729:Terpenes; C058582:beta-phellandrene; C004809:isopentenyl pyrophosphate; C043060:3,3-dimethylallyl pyrophosphate; D002244:Carbon; D004896:Erythritol; D008798:Mevalonic Acid