An efficient xylose-fermenting recombinant Saccharomyces cerevisiae strain obtained through adaptive evolution and its global transcription profile
Factors related to ethanol production from xylose in engineered Saccharomyces cerevisiae that contain an exogenous initial metabolic pathway are still to be elucidated. In the present study, a strain that expresses the xylose isomerase gene of Piromyces sp. Pi-xylA and overexpresses XKS1, RPE1, RKI1, TAL1, and TKL1, with deleted GRE3 and COX4 genes was constructed. The xylose utilization capacity of the respiratory deficiency strain was poor but improved via adaptive evolution in xylose. The μ max of the evolved strain in 20 g l−1 xylose is 0.11 ± 0.00 h−1, and the evolved strain consumed 17.83 g l−1 xylose within 72 h, with an ethanol yield of 0.43 g g−1 total consumed sugars during glucose–xylose cofermentation. Global transcriptional changes and effect of several specific genes were studied. The result revealed that the increased xylose isomerase acivity, the upregulation of enzymes involved in glycolysis and glutamate synthesis, and the downregulation of trehalose and glycogen synthesis, may have contributed to the improved xylose utilization of the strain. Furthermore, the deletion of PHO13 decreased the xylose growth in the respiration deficiency strain although deleting PHO13 can improve the xylose metabolism in other strains.
KeywordsXylose isomerase Respiratory deficiency Ethanol PDC6 PHO13 CWP1
This work was supported by the National Key Basic Research Program (2011CB707405), the International S&T Cooperation Program of China (2010DFA32560), and the National Natural Science Foundation of China (30970091 and 31070096).We would like to thank Dr. Peter Kötter of the Johann Wolfgang Goethe-University in Frankfurt for supplying the CEN.PK strains.
- Entian KD, Kötter P (1998) Yeast mutant and plasmid collections. In: Brown AJP, Tuite MF (eds) Methods in microbiology, volume 26. Academic, San Diego, pp 431–449Google Scholar
- Garcia Sanchez R, Karhumaa K, Fonseca C, Sanchez Nogue V, Almeida JR, Larsson CU, Bengtsson O, Bettiga M, Hahn-Hägerdal B, Gorwa-Grauslund MF (2010) Improved xylose and arabinose utilization by an industrial recombinant Saccharomyces cerevisiae strain using evolutionary engineering. Biotechnol Biofuels 3:13CrossRefGoogle Scholar
- Hohmann S (1991) Characterization of PDC6, a third structural gene for pyruvate decarboxylase in Saccharomyces cerevisiae. J Bacteriol 173(24):7963–7969Google Scholar
- Lee P, Cho BR, Joo HS, Hahn JS (2008) Yeast Yak1 kinase, a bridge between PKA and stress-responsive transcription factors, Hsf1 and Msn2/Msn4. Mol Microbiol 70(4):882–895Google Scholar
- Peng B, Chen X, Shen Y, Bao X (2011) Effect of controlled overexpression of xylulokinase by different promoters on xylose metabolism in Saccharomyces cerevisiae. Wei Sheng Wu Xue Bao 51(7):914–922Google Scholar
- Sauer U (2001) Evolutionary engineering of industrially important microbial phenotypes. Adv Biochem Eng Biotechnol 73:129–169Google Scholar
- van Maris AJ, Abbott DA, Bellissimi E, van den Brink J, Kuyper M, Luttik MA, Wisselink HW, Scheffers WA, van Dijken JP, Pronk JT (2006) Alcoholic fermentation of carbon sources in biomass hydrolysates by Saccharomyces cerevisiae: current status. Antonie Van Leeuwenhoek 90(4):391–418CrossRefGoogle Scholar
- Wohlbach DJ, Kuo A, Sato TK, Potts KM, Salamov AA, Labutti 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 (2011) Comparative genomics of xylose-fermenting fungi for enhanced biofuel production. Proc Natl Acad Sci U S A 108(32):13212–13217CrossRefGoogle Scholar