Ahsan Islam (4116055)

Barbara Bourgade (5472902)

Brahim Benyahia (1258056)

James Millar (8881445)

Nigel Minton (565961)

English

Loughborough University Institutional Repository

Implementation of Synthetic Metabolic Pathways in Moorellathermoacetica and Clostridium autoethanogenumBarbara Bourgade1,2, James Millard2, Brahim Benyahia1, Nigel Minton2, M. Ahsanul Islam11Department of Chemical Engineering, Loughborough University 2Synthetic Biology Research Centre, University of Nottingham• Microbial chassis organisms are crucial forchemicals bioproduction.• Acetogenic bacteria, or acetogens, arepromising hosts as they convert CO and CO2into acetate and other products during gasfermentation.CONTACT INFORMATIONDepartment of Chemical EngineeringLoughborough University Leicestershire LE11 3TU  United Kingdomb.bourgade@lboro.ac.uk  REFERENCE1 Chou et al (2009). Nucleic Acid Research (37)2 Kuwahara et al (2016). Nucleic Acids Research (44)3 Becker et al (2007). Nature Protocols (2)ACKNOWLEDGEMENTS • Loughborough University • Prof Nigel Minton, Dr James Millard and the Synthetic Biology Research Centre (Nottingham) for their help with the experimental part IntroductionCO2CO2 fixation by acetogensCOWOOD-LJUNGDAHL PATHWAYAcetyl-CoAAcetate BiomassPathway implementation in C. autoethanogenum• Synthetic metabolic pathways for ethylene glycol and1,2-propanediol designed using the computationaltools From Metabolite to Metabolite1 and MetabolicRoute Explorer2• Further analysis with the constraint-based COBRA3toolbox.Future workGenetic tools for M. thermoacetica• Pathway expression in Clostridiumautoethanogenum with an inducible system• No production detected• Addition of a FLAG-tag to the enzymes to assessexpression• Moorella thermoacetica: several industrial advantages• Limited genetic tools available• Several challenges complicate successful DNAtransfer• Shuttle vector built• Compatible gram-positive replicon identified• Successful transformation• Mixed population: plasmid instability?• Increase of plasmid stability• Alternative selection marker; toxin-antitoxin system• Development of reporter system to test promoteractivity• The thermophilic acetogen Moorellathermoacetica has an interesting industrialpotential but better genetic tools are required tounlock this potential.• Computationally designed synthetic pathways forchemicals production are also currently beingimplemented in another acetogen, Clostridiumautoethanogenum.• Different aspects to consider for target production• Alternative expression system• Cofactor balance• Enzyme compatibility• Increase plasmid stability• Identify promoters• Apply developed genetic tools

Implementation of synthetic metabolic pathways in Moorella thermoacetica and Clostridium autoethanogenum

https://repository.lboro.ac.uk/articles/Implementation_of_synthetic_metabolic_pathways_in_Moorella_thermoacetica_and_Clostridium_autoethanogenum/12357863/files/22775333.pdf

Implementation of synthetic metabolic pathways in Moorella thermoacetica and Clostridium autoethanogenum

Abstract

Similar works

Full text

Available Versions

Loughborough University Institutional Repository