期刊论文详细信息
Biotechnology for Biofuels
Improved ethanol production by a xylose-fermenting recombinant yeast strain constructed through a modified genome shuffling method
Anli Geng1  Wei Zhang1 
[1]School of Life Sciences and Chemical Technology, Ngee Ann Polytechnic, 535 Clementi Road, Singapore, 599489, Singapore
关键词: Cellulosic ethanol;    Xylose;    Genome shuffling;    Yeast;    P. stipitis;    S. cerevisiae;   
Others  :  798269
DOI  :  10.1186/1754-6834-5-46
 received in 2011-11-02, accepted in 2012-01-11,  发布年份 2012
PDF
【 摘 要 】

Background

Xylose is the second most abundant carbohydrate in the lignocellulosic biomass hydrolysate. The fermentation of xylose is essential for the bioconversion of lignocelluloses to fuels and chemicals. However the wild-type strains of Saccharomyces cerevisiae are unable to utilize xylose. Many efforts have been made to construct recombinant yeast strains to enhance xylose fermentation over the past few decades. Xylose fermentation remains challenging due to the complexity of lignocellulosic biomass hydrolysate. In this study, a modified genome shuffling method was developed to improve xylose fermentation by S. cerevisiae. Recombinant yeast strains were constructed by recursive DNA shuffling with the recombination of entire genome of P. stipitis with that of S. cerevisiae.

Results

After two rounds of genome shuffling and screening, one potential recombinant yeast strain ScF2 was obtained. It was able to utilize high concentration of xylose (100 g/L to 250 g/L xylose) and produced ethanol. The recombinant yeast ScF2 produced ethanol more rapidly than the naturally occurring xylose-fermenting yeast, P. stipitis, with improved ethanol titre and much more enhanced xylose tolerance.

Conclusion

The modified genome shuffling method developed in this study was more effective and easier to operate than the traditional protoplast-fusion-based method. Recombinant yeast strain ScF2 obtained in this study was a promising candidate for industrial cellulosic ethanol production. In order to further enhance its xylose fermentation performance, ScF2 needs to be additionally improved by metabolic engineering and directed evolution.

【 授权许可】

   
2012 Zhang and Geng; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20140706113831424.pdf 1053KB PDF download
Figure 6 . 38KB Image download
Figure 5 . 49KB Image download
Figure 4 . 18KB Image download
Figure 3 . 54KB Image download
Figure 2 . 28KB Image download
Figure 1 . 23KB Image download
【 图 表 】

Figure 1 .

Figure 2 .

Figure 3 .

Figure 4 .

Figure 5 .

Figure 6 .

【 参考文献 】
  • [1]Jeffries TW, Jin YS: Metabolic engineering for improved fermentation of pentoses by yeasts. Appl Microbiol Biotechnol 2004, 63:495-509.
  • [2]Jeppsson M, Traff K, Johansson B, Hahn-Hagerdal B, Gorwa-Grauslund MF: Effect of enhanced xylose reductase activity on xylose consumption and product distribution in xylose-fermenting recombinant Saccharomyces cerevisiae. FEMS Yeast Res 2002, 3:167-175.
  • [3]Pasha C, Kuhad RC, Venkateswar Rao L: Stain improvement of thermotolerant Saccharomyces cerevisiae VS3 strain for better utilization of lignocellulosic substrates. J Appl Microbiol 2007, 103:1480-1489.
  • [4]Ho NW, Chen Z, Brainard AP: Genetically engineered Saccharomyces yeast capable of effective co-fermentation of glucose and xylose. Appl Environ Microbiol 1998, 64(5):1852-1859.
  • [5]Gong JX, Zheng HJ, Wu ZJ, Chen T, Zhao XM: Genome shuffling: progress and applications for phenotype improvement. Biotechnol Adv 2009, 27(6):996-1005.
  • [6]Ness JE, Welch M, Giver L, Bueno M, Cherry JR, Borchert TV, Stemmer WPC, Minshull J: DNA shuffling of subgenomic sequences of subtilisin. Nat Biotechnol 1999, 17:893-896.
  • [7]Patnaik R, Louie S, Gavrilovic V, Stemmer WPC, Ryan CM, Cardayre S: Genome shuffling of lactobacillus for improved acid tolerance. Nat Biotechnol 2002, 20:707-712.
  • [8]Dai MH, Copley SD: Genome shuffling improves degradation of the anthropogenic pesticide pentachlorophenol by Sphingobium chlorophenolicum ATCC 39723. Appl Environ Microbiol 2004, 70:2391-2397.
  • [9]Hou L: Improved production of ethanol by novel genome shuffling in Saccharomyces cerevisiae. Appl Biochem Biotechnol 2010, 160:1084-1093.
  • [10]Wohlbach DJ, Kuo A, Sato TK, Potts KM, Salamov AA, LaButte KM, Sun H, Clum A, Pangilinan JL, Lindquist EA, Lucas S, Lapidus A, Jin M, Gunawan C, Balan V, Dale BE, Jeffries TW, Zinkel R, Barry KW, Grigoriev IV, Gasch AP: Comparative genomics of xylose-fermenting fungi for enhanced biofuel production. PNAS 2011.
  • [11]Slininger PJ, Thompson SR, Weber S, Liu ZL, Moon J: Repression of xylose-specific enzymes by ethanol in Scheffersomyces (Pichia) stipitis and utility of repitching xylose-grown populations to eliminate diauxic lag. Biotechnol Bioeng 2011, 108(8):1801-1815.
  • [12]Lin Y, Tanaka S: Ethanol fermentation from biomass resources: current state and prospects. Appl Microbiol Biotechnol 2005, 69:627-642.
  • [13]Eliasson A, Christensson C, Wahlbom CF, Hahn-Hägerdal B: Anaerobic xylose fermentation by recombinant Saccharomyces cerevisiae carrying XYL1, XYL2, and XKS1 in mineral medium chemostat cultures. Appl Environ Microbiol 2000, 66(8):3381-3386.
  • [14]Toivari MH, Aristidou A, Ruohonen L, Penttilä M: Conversion of xylose to ethanol by recombinant Saccharomyces cerevisiae: importance of xylulokinase (XKS1) and oxygen availability. Metab Eng 2001, 3(3):236-249.
  • [15]Gupthar AS: Segregation of altered parental properties in fusions between Saccharomyces cerevisiae and the D-xylose fermenting yeasts Candida shehatae and Pichia stipitis. Can J Microbiol 1992, 38(12):1233-1237.
  • [16]Kordowska-Wiater M, Targonski Z: Application of Saccharomyces cerevisiae and Pichia stipitis karyoductants to the production of ethanol from xylose. Acta Microbiol Pol 2001, 50(3–4):291-299.
  • [17]Yoon G-S, Lee T-S, Kim C, Seo J-H, Ryu Y-W: Characterization of alcohol fermentation and segregation of protoplast fusant of Saccharomyces cerevisiae and Pichia stipitis. J Microbiol Biotechnol 1996, 6(4):286-291.
  • [18]Welsh J, McCleland M: Genomic fingerprinting using arbitrarily primed PCR and a matrix of pairwise combinations of primers. Nucleic Acids Res 1991, 19(19):5275-5279.
  • [19]Williams JGK, Kubelik AR, Livak KJ, Rafalski JA: DNA polymorfisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 1990, 18(22):6531-6535.
  • [20]Heluane H, Defigueroa LIC, Vázquez F: Fusion of yeast protoplasts and isolated nuclei of Fusarium moniliforme. Acta Biotechnol 1998, 18(4):353-359.
  • [21]Valério HM, Weikert-Oliveira RCB, Resende MA: Differentiation of Candida species obtained from nosocomial candidemia using RAPD-PCR technique. Rev Soc Bras Med Trop 2006, 39:174-178.
  文献评价指标  
  下载次数:83次 浏览次数:24次