Biotechnology for Biofuels | |
A microbial platform for renewable propane synthesis based on a fermentative butanol pathway | |
Navya Menon2  András Pásztor3  Binuraj RK Menon2  Pauli Kallio3  Karl Fisher2  M Kalim Akhtar3  David Leys2  Patrik R Jones1  Nigel S Scrutton2  | |
[1] Department of Life Sciences, Imperial College London, Sir Alexander Fleming Building, South Kensington Campus, London SW7 2AZ, UK | |
[2] BBSRC/EPSRC Centre for Synthetic Biology of Fine and Speciality Chemicals, Manchester Institute of Biotechnology, Faculty of Life Sciences, 131 Princess Street, The University of Manchester, Manchester M1 7DN, UK | |
[3] Molecular Plant Biology, Department of Biochemistry, Tykistökatu 6A 6krs, University of Turku, FI 20014 TURUN YLIOPISTO, Turku, Finland | |
关键词: Escherchia coli; Aldehyde deformylating oxygenase; Cyanobacteria; Microbial pathway engineering; Butanol; Propane; | |
Others : 1177293 DOI : 10.1186/s13068-015-0231-1 |
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received in 2014-11-10, accepted in 2015-02-23, 发布年份 2015 | |
【 摘 要 】
Background
Propane (C3H8) is a volatile hydrocarbon with highly favourable physicochemical properties as a fuel, in addition to existing global markets and infrastructure for storage, distribution and utilization in a wide range of applications. Consequently, propane is an attractive target product in research aimed at developing new renewable alternatives to complement currently used petroleum-derived fuels. This study focuses on the construction and evaluation of alternative microbial biosynthetic pathways for the production of renewable propane. The new pathways utilize CoA intermediates that are derived from clostridial-like fermentative butanol pathways and are therefore distinct from the first microbial propane pathways recently engineered in Escherichia coli.
Results
We report the assembly and evaluation of four different synthetic pathways for the production of propane and butanol, designated a) atoB-adhE2 route, b) atoB-TPC7 route, c) nphT7-adhE2 route and d) nphT7-TPC7 route. The highest butanol titres were achieved with the atoB-adhE2 (473 ± 3 mg/L) and atoB-TPC7 (163 ± 2 mg/L) routes. When aldehyde deformylating oxygenase (ADO) was co-expressed with these pathways, the engineered hosts also produced propane. The atoB-TPC7-ADO pathway was the most effective in producing propane (220 ± 3 μg/L). By (i) deleting competing pathways, (ii) including a previously designed ADOA134F variant with an enhanced specificity towards short-chain substrates and (iii) including a ferredoxin-based electron supply system, the propane titre was increased (3.40 ± 0.19 mg/L).
Conclusions
This study expands the metabolic toolbox for renewable propane production and provides new insight and understanding for the development of next-generation biofuel platforms. In developing an alternative CoA-dependent fermentative butanol pathway, which includes an engineered ADO variant (ADOA134F), the study addresses known limitations, including the low bio-availability of butyraldehyde precursors and poor activity of ADO with butyraldehyde.
【 授权许可】
2015 Menon et al.; licensee BioMed Central.
【 预 览 】
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【 参考文献 】
- [1]Hedley M, Jiang W, McLaren R, Singleton DL: Modeling of future-year emissions control scenarios for the lower Fraser Valley: impacts of natural gas and propane vehicle technologies. J Appl Meteorol 1998, 37:1190-204.
- [2]Shelley C: The story of LPG. Poten and Partners, London; 2003.
- [3]Laciak M: Properties of artificial gaseous mixtures for their safe use and support the natural gas supply networks. Arch Min Sci 2012, 57:351-62.
- [4]Thorsteinsson HH, Tester JW: Barriers and enablers to geothermal district heating system development in the United States. Energ Policy 2010, 38:803-13.
- [5]Keller FJ, Liang HM, Farzad M: Assessment of propane as a refrigerant in residential air-conditioning and heat pump applications. Refrigerants for the 21st Century. 1997.
- [6]Velasco I, Rivas C, Martinez-Lopez JF, Blanco ST, Otin S, Artal M: Accurate values of some thermodynamic properties for carbon dioxide, ethane, propane, and some binary mixtures. J Phys Chem B 2011, 115:8216-30.
- [7]Schirmer A, Rude MA, Li XZ, Popova E, del Cardayre SB: Microbial biosynthesis of alkanes. Science 2010, 329:559-62.
- [8]Akhtar MK, Turner NJ, Jones PR: Carboxylic acid reductase is a versatile enzyme for the conversion of fatty acids into fuels and chemical commodities. Proc Natl Acad Sci U S A 2013, 110:87-92.
- [9]Howard TP, Middelhaufe S, Moore K, Edner C, Kolak DM, Taylor GN, et al.: Synthesis of customized petroleum-replica fuel molecules by targeted modification of free fatty acid pools in Escherichia coli. Proc Natl Acad Sci U S A 2013, 110:7636-41.
- [10]Kallio P, Pasztor A, Thiel K, Akhtar MK, Jones PR: An engineered pathway for the biosynthesis of renewable propane. Nat Commun 2014, 5:4731.
- [11]Lang K, Zierow J, Buehler K, Schmid A: Metabolic engineering of Pseudomonas sp. strain VLB120 as platform biocatalyst for the production of isobutyric acid and other secondary metabolites. Microb Cell Fact 2014, 13:2. BioMed Central Full Text
- [12]Khara B, Menon N, Levy C, Mansell D, Das D, Marsh ENG, et al.: Production of propane and other short-chain alkanes by structure-based engineering of ligand specificity in aldehyde-deformylating oxygenase. Chembiochem 2013, 14:1204-8.
- [13]Coates RC, Podell S, Korobeynikov A, Lapidus A, Pevzner P, Sherman DH, et al.: Characterization of cyanobacterial hydrocarbon composition and distribution of biosynthetic pathways. Plos One 2014, 9:e85140.
- [14]Atsumi S, Higashide W, Liao JC: Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde. Nat Biotechnol 2009, 27:1177-80.
- [15]Bond-Watts BB, Bellerose RJ, Chang MCY: Enzyme mechanism as a kinetic control element for designing synthetic biofuel pathways. Nat Chem Biol 2011, 7:222-7.
- [16]Dellomonaco C, Clomburg JM, Miller EN, Gonzalez R: Engineered reversal of the beta-oxidation cycle for the synthesis of fuels and chemicals. Nature 2011, 476:355-U131.
- [17]Lan EI, Liao JC: Metabolic engineering of cyanobacteria for 1-butanol production from carbon dioxide. Metab Eng 2011, 13:353-63.
- [18]Lan EI, Liao JC: ATP drives direct photosynthetic production of 1-butanol in cyanobacteria. Proc Natl Acad Sci U S A 2012, 109:6018-23.
- [19]Lan EI, Ro SY, Liao JC: Oxygen-tolerant coenzyme A-acylating aldehyde dehydrogenase facilitates efficient photosynthetic n-butanol biosynthesis in cyanobacteria. Energ Environ Sci 2013, 6:2672-81.
- [20]Pasztor A, Kallio P, Malatinszky D, Akhtar MK, Jones PR: A synthetic O-tolerant butanol pathway exploiting native fatty acid biosynthesis in Escherichia coli. Biotechnol Bioeng 2015, 112:120-8.
- [21]Shen CR, Lan EI, Dekishima Y, Baez A, Cho KM, Liao JC: Driving forces enable high-titer anaerobic 1-butanol synthesis in Escherichia coli. Appl Environ Microb 2011, 77:2905-15.
- [22]Fontaine L, Meynial-Salles I, Girbal L, Yang XH, Croux C, Soucaille P: Molecular characterization and transcriptional analysis of adhE2, the gene encoding the NADH-dependent aldehyde/alcohol dehydrogenase responsible for butanol production in alcohologenic cultures of Clostridium acetobutylicum ATCC 824. J Bacteriol 2002, 184:821-30.
- [23]Zhuang ZH, Song F, Zhao H, Li L, Cao J, Eisenstein E, et al.: Divergence of function in the hot dog fold enzyme superfamily: the bacterial thioesterase YciA. Biochemistry-Us 2008, 47:2789-96.
- [24]Rodriguez GM, Atsumi S: Toward aldehyde and alkane production by removing aldehyde reductase activity in Escherichia coli. Metab Eng 2014, 25:227-37.
- [25]Davis MS, Cronan JE: Inhibition of Escherichia coli acetyl coenzyme A carboxylase by acyl-acyl carrier protein. J Bacteriol 2001, 183:1499-503.
- [26]James ES, Cronan JE: Expression of two Escherichia coli acetyl-CoA carboxylase subunits is autoregulated. J Biol Chem 2004, 279:2520-7.
- [27]Yee BC, Delatorre A, Crawford NA, Lara C, Carlson DE, Buchanan BB: The ferredoxin-thioredoxin system of enzyme regulation in a cyanobacterium. Arch Microbiol 1981, 130:14-8.
- [28]Buckel W, Thauer RK: Energy conservation via electron bifurcating ferredoxin reduction and proton/Na + translocating ferredoxin oxidation. Biochimica Et Biophysica Acta-Bioenergetics 2013, 1827:94-113.
- [29]Zhang JJ, Lu XF, Li JJ: Conversion of fatty aldehydes into alk (a/e)nes by in vitro reconstituted cyanobacterial aldehyde-deformylating oxygenase with the cognate electron transfer system. Biotechnol Biofuels 2013, 6:86. BioMed Central Full Text
- [30]Eser BE, Das D, Han J, Jones PR, Marsh ENG: Oxygen-independent alkane formation by non-heme iron-dependent cyanobacterial aldehyde decarbonylase: investigation of kinetics and requirement for an external electron donor. Biochemistry-US 2011, 50:10743-50.