| Biotechnology for Biofuels | |
| Butanol production in S. cerevisiae via a synthetic ABE pathway is enhanced by specific metabolic engineering and butanol resistance | |
| R. Swidah1  H. Wang1  P.J. Reid1  H.Z. Ahmed1  A.M. Pisanelli2  K.C. Persaud2  C.M. Grant1  M.P. Ashe1  | |
| [1] The Faculty of Life Sciences, The Michael Smith Building, The University of Manchester, Oxford Rd., Manchester M13 9PT, UK | |
| [2] School of Chemical engineering and Analytical Science, The Mill, The University of Manchester, Sackville St., Manchester M139PL, UK | |
| 关键词: ABE pathway; Saccharomyces cerevisiae; Biobutanol; | |
| Others : 1219144 DOI : 10.1186/s13068-015-0281-4 |
|
| received in 2015-03-31, accepted in 2015-06-29, 发布年份 2015 | |
PDF
|
|
【 摘 要 】
Background
The fermentation of sugars to alcohols by microbial systems underpins many biofuel initiatives. Short chain alcohols, like n-butanol, isobutanol and isopropanol, offer significant advantages over ethanol in terms of fuel attributes. However, production of ethanol from resistant Saccharomyces cerevisiae strains is significantly less complicated than for these alternative alcohols.
Results
In this study, we have transplanted an n-butanol synthesis pathway largely from Clostridial sp. to the genome of an S. cerevisiae strain. Production of n-butanol is only observed when additional genetic manipulations are made to restore any redox imbalance and to drive acetyl-CoA production. We have used this butanol production strain to address a key question regarding the sensitivity of cells to short chain alcohols. In the past, we have defined specific point mutations in the translation initiation factor eIF2B based upon phenotypic resistance/sensitivity to high concentrations of exogenously added n-butanol. Here, we show that even during endogenous butanol production, a butanol resistant strain generates more butanol than a butanol sensitive strain.
Conclusion
These studies demonstrate that appreciable levels of n-butanol can be achieved in S. cerevisiae but that significant metabolic manipulation is required outside of the pathway converting acetyl-CoA to butanol. Furthermore, this work shows that the regulation of protein synthesis by short chain alcohols in yeast is a critical consideration if higher yields of these alcohols are to be attained.
【 授权许可】
2015 Swidah et al.
【 预 览 】
| Files | Size | Format | View |
|---|---|---|---|
| 20150715080754114.pdf | 1898KB | ||
| Fig. 5. | 45KB | Image | |
| Fig. 4. | 35KB | Image | |
| Fig. 3. | 30KB | Image | |
| Fig. 2. | 54KB | Image | |
| Fig. 1. | 58KB | Image |
【 图 表 】
Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
【 参考文献 】
- [1]Kerr RA: Climate change. Global warming is changing the world. Science 2007, 316:188-90.
- [2]Peralta-Yahya PP, Zhang F, del Cardayre SB, Keasling JD: Microbial engineering for the production of advanced biofuels. Nature. 2012, 488:320-8.
- [3]Laluce C, Schenberg AC, Gallardo JC, Coradello LF, Pombeiro-Sponchiado SR: Advances and developments in strategies to improve strains of Saccharomyces cerevisiae and processes to obtain the lignocellulosic ethanol—a review. Appl Biochem Biotechnol. 2012, 166:1908-26.
- [4]Cascone R: Biobutanol—a replacement for bioethanol? Chem Eng Prog. 2008, 104:S4-9.
- [5]Zheng YN, Li LZ, Xian M, Ma YJ, Yang JM, Xu X, et al.: Problems with the microbial production of butanol. J Ind Microbiol Biotechnol. 2009, 36:1127-38.
- [6]Bud R: The uses of life: a history of biotechnology. Cambridge University Press, Cambridge; 1993.
- [7]Jang YS, Lee J, Malaviya A, Seung do Y, Cho JH, Lee SY: Butanol production from renewable biomass: rediscovery of metabolic pathways and metabolic engineering. Biotechnol J 2012, 7:186-98.
- [8]Xue C, Zhao XQ, Liu CG, Chen LJ, Bai FW: Prospective and development of butanol as an advanced biofuel. Biotechnol Adv. 2013, 31:1575-84.
- [9]Hong KK, Nielsen J: Metabolic engineering of Saccharomyces cerevisiae: a key cell factory platform for future biorefineries. Cell Mol Life Sci. 2012, 69:2671-90.
- [10]Lan EI, Liao JC: Microbial synthesis of n-butanol, isobutanol, and other higher alcohols from diverse resources. Bioresour Technol. 2013, 135:339-49.
- [11]Bond-Watts BB, Bellerose RJ, Chang MC: Enzyme mechanism as a kinetic control element for designing synthetic biofuel pathways. Nat Chem Biol. 2011, 7:222-7.
- [12]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 Microbiol. 2011, 77:2905-15.
- [13]Huffer S, Roche CM, Blanch HW, Clark DS: Escherichia coli for biofuel production: bridging the gap from promise to practice. Trends Biotechnol. 2012, 30:538-45.
- [14]Generoso WC, Schadeweg V, Oreb M, Boles E: Metabolic engineering of Saccharomyces cerevisiae for production of butanol isomers. Curr Opin Biotechnol. 2014, 33C:1-7.
- [15]Steen EJ, Chan R, Prasad N, Myers S, Petzold CJ, Redding A, et al.: Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol. Microb Cell Fact. 2008, 7:36. BioMed Central Full Text
- [16]Sakuragi H, Morisaka H, Kuroda K, Ueda M: Enhanced butanol production by eukaryotic Saccharomyces cerevisiae engineered to contain an improved pathway. Biosci Biotechnol Biochem 2015, 79:314-20.
- [17]Krivoruchko A, Serrano-Amatriain C, Chen Y, Siewers V, Nielsen J: Improving biobutanol production in engineered Saccharomyces cerevisiae by manipulation of acetyl-CoA metabolism. J Ind Microbiol Biotechnol. 2013, 40:1051-6.
- [18]Si T, Luo Y, Xiao H, Zhao H: Utilizing an endogenous pathway for 1-butanol production in Saccharomyces cerevisiae. Metab Eng. 2014, 22:60-8.
- [19]Branduardi P, Longo V, Berterame NM, Rossi G, Porro D: A novel pathway to produce butanol and isobutanol in Saccharomyces cerevisiae. Biotechnol Biofuels. 2013, 6:68. BioMed Central Full Text
- [20]Ashe MP, Slaven JW, De Long SK, Ibrahimo S, Sachs AB: A novel eIF2B-dependent mechanism of translational control in yeast as a response to fusel alcohols. Embo J. 2001, 20:6464-74.
- [21]Taylor EJ, Campbell SG, Griffiths CD, Reid PJ, Slaven JW, Harrison RJ, et al.: Fusel alcohols regulate translation initiation by inhibiting eIF2B to reduce ternary complex in a mechanism that may involve altering the integrity and dynamics of the eIF2B body. Mol Biol Cell. 2010, 21:2202-16.
- [22]Smirnova JB, Selley JN, Sanchez-Cabo F, Carroll K, Eddy AA, McCarthy JE, et al.: Global gene expression profiling reveals widespread yet distinctive translational responses to different eukaryotic translation initiation factor 2B-targeting stress pathways. Mol Cell Biol. 2005, 25:9340-9.
- [23]Egbe NE, Paget CM, Wang H, Ashe MP: Alcohols inhibit translation to regulate morphogenesis in C. albicans. Fungal Genet Biol 2015, 77:50-60.
- [24]Pavitt GD: eIF2B, a mediator of general and gene-specific translational control. Biochem Soc Trans. 2005, 33:1487-92.
- [25]Flagfeldt DB, Siewers V, Huang L, Nielsen J: Characterization of chromosomal integration sites for heterologous gene expression in Saccharomyces cerevisiae. Yeast. 2009, 26:545-51.
- [26]Paquin CE, Williamson VM: Ty insertions at two loci account for most of the spontaneous antimycin A resistance mutations during growth at 15 degrees C of Saccharomyces cerevisiae strains lacking ADH1. Mol Cell Biol. 1986, 6:70-9.
- [27]Slavov N, Botstein D: Coupling among growth rate response, metabolic cycle, and cell division cycle in yeast. Mol Biol Cell. 2011, 22:1997-2009.
- [28]Dunlop MJ, Dossani ZY, Szmidt HL, Chu HC, Lee TS, Keasling JD, et al.: Engineering microbial biofuel tolerance and export using efflux pumps. Mol Syst Biol. 2011, 7:487.
- [29]Guthrie C, Fink GR: Guide to yeast genetics and molecular biology. Academic, San Diego, California; 1991.
- [30]Ashe MP, De Long SK, Sachs AB: Glucose depletion rapidly inhibits translation initiation in yeast. Mol Biol Cell. 2000, 11:833-48.
PDF