Engineering the Autotroph Methanococcus maripaludis for Geraniol Production

ACS Synth Biol. 2016 Jul 15;5(7):577-81. doi: 10.1021/acssynbio.5b00267. Epub 2016 Feb 17.

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.

Keywords: archaea; carbon dioxide; hydrogen economy; isoprenoid; methanogen; synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyl Coenzyme A / metabolism
  • Acyclic Monoterpenes
  • Autotrophic Processes
  • Formates / metabolism
  • Genetic Engineering / methods*
  • Methanococcus / drug effects
  • Methanococcus / genetics
  • Methanococcus / metabolism*
  • Ocimum basilicum / genetics
  • Phosphoric Monoester Hydrolases / genetics
  • Phosphoric Monoester Hydrolases / metabolism
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Puromycin / pharmacology
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Terpenes / metabolism*

Substances

  • Acyclic Monoterpenes
  • Formates
  • Plant Proteins
  • Recombinant Proteins
  • Terpenes
  • formic acid
  • Puromycin
  • Acetyl Coenzyme A
  • Phosphoric Monoester Hydrolases
  • geraniol