Metabolic engineering of Cupriavidus necator for heterotrophic and autotrophic alka(e)ne production |
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| Institution: | 1. INRA, UMR792, Ingénierie des Systèmes Biologiques et des Procédés, Toulouse, France;2. CNRS, UMR5504, 135 avenue de rangueil, F-31400 Toulouse, France;3. CNRS; LAAS; 7 avenue du Colonel Roche, F-31400 Toulouse, France;4. Université de Toulouse; UPS, LAAS; F-31400 Toulouse, France;5. Université de Toulouse; INSA, LISBP; F-31400 Toulouse, France;6. Department of Biology, Massachusetts Institute of Technology, Bldg. 68-370, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA;7. Division of Health Sciences and Technology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA;8. Engineering Systems Division, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA;1. Flemish Institute for Technological Research (VITO), Business Unit Separation and Conversion Technology, Boeretang 200, 2400 Mol, Belgium;2. Ghent University, Department of Biosystems Engineering, Coupure Links 653, 9000 Ghent, Belgium;1. CAS Key Laboratory of Bio-based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, 266101 Qingdao, China;2. Mansoura University, Faculty of Agriculture, 35516 Mansoura, Egypt;3. Aachen University of Applied Sciences, Campus Juelich, Department of Chemistry and Biotechnology, Heinrich-Mussmann-Str.1, D-52428 Juelich, Germany;4. College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266042, China;1. Institute for Applied Biosciences, Department of Applied Biology, Karlsruhe Institute of Technology, Karlsruhe, Germany;2. University Grenoble Alpes, CEA, INSERM, IRIG, BGE, Grenoble, France;3. Institute for Biological Interfaces, Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany |
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| Abstract: | Alkanes of defined carbon chain lengths can serve as alternatives to petroleum-based fuels. Recently, microbial pathways of alkane biosynthesis have been identified and enabled the production of alkanes in non-native producing microorganisms using metabolic engineering strategies. The chemoautotrophic bacterium Cupriavidus necator has great potential for producing chemicals from CO2: it is known to have one of the highest growth rate among natural autotrophic bacteria and under nutrient imbalance it directs most of its carbon flux to the synthesis of the acetyl-CoA derived polymer, polyhydroxybutyrate (PHB), (up to 80% of intracellular content). Alkane synthesis pathway from Synechococcus elongatus (2 genes coding an acyl-ACP reductase and an aldehyde deformylating oxygenase) was heterologously expressed in a C. necator mutant strain deficient in the PHB synthesis pathway. Under heterotrophic condition on fructose we showed that under nitrogen limitation, in presence of an organic phase (decane), the strain produced up to 670 mg/L total hydrocarbons containing 435 mg/l of alkanes consisting of 286 mg/l of pentadecane, 131 mg/l of heptadecene, 18 mg/l of heptadecane, and 236 mg/l of hexadecanal. We report here the highest level of alka(e)nes production by an engineered C. necator to date. We also demonstrated the first reported alka(e)nes production by a non-native alkane producer from CO2 as the sole carbon source. |
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| Keywords: | Alkane Alkene Hydrocarbon Biofuels Metabolic engineering Fermentation |
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