title: Engineering the Autotroph Methanococcus maripaludis for Geraniol Production.
abstract: The rapid autotrophic growth of the methanogenic archaeon Methanococcus maripaludis on H2 and CO2 makes it an attractive microbial chassis to inexpensively produce biochemicals. To explore this potential, a synthetic gene encoding geraniol synthase (GES) derived from Ocimum basilicum was cloned into a M. maripaludis expression vector under selection for puromycin resistance. Recombinant expression of GES in M. maripaludis during autotrophic growth on H2/CO2 or formate yielded geraniol at 2.8 and 4.0 mg g(-1) of dry weight, respectively. The yield of geraniol decreased 2-3-fold when organic carbon sources were added to stimulate heterotrophic growth. In the absence of puromycin, geraniol production during autotrophic growth on formate increased to 4.6 mg g(-1) of dry weight. A conceptual model centered on the autotrophic acetyl coenzyme A biosynthetic pathway identified strategies to divert more autotrophic carbon flux to geraniol production.
authors.affiliation: Department of Microbiology, ‡College of Engineering, and §Department of Biochemistry and Molecular Biology, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Lyu
authors.affiliation: Department of Microbiology, ‡College of Engineering, and §Department of Biochemistry and Molecular Biology, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: Rachit
authors.initials: R
authors.lastname: Jain
authors.affiliation: Department of Microbiology, ‡College of Engineering, and §Department of Biochemistry and Molecular Biology, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: Peyton
authors.initials: P
authors.lastname: Smith
authors.affiliation: Department of Microbiology, ‡College of Engineering, and §Department of Biochemistry and Molecular Biology, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: Travis
authors.initials: T
authors.lastname: Fetchko
authors.affiliation: Department of Microbiology, ‡College of Engineering, and §Department of Biochemistry and Molecular Biology, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Microbiology, ‡College of Engineering, and §Department of Biochemistry and Molecular Biology, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: William B
authors.initials: WB
authors.lastname: Whitman
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism; D000080462:Acyclic Monoterpenes; D052818:Autotrophic Processes; D005561:Formates / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D017017:Methanococcus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018649:Ocimum basilicum / Q000235:genetics; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D011691:Puromycin / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism*
chemical_list: D000080462:Acyclic Monoterpenes; D005561:Formates; D010940:Plant Proteins; D011994:Recombinant Proteins; D013729:Terpenes; C030544:formic acid; D011691:Puromycin; D000105:Acetyl Coenzyme A; D010744:Phosphoric Monoester Hydrolases; C007836:geraniol
keywords: archaea; carbon dioxide; hydrogen economy; isoprenoid; methanogen; synthetic biology