Key Laboratory of Systems Bioengineering, Ministry of Education; Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, People's Republic of China.
PLoS One. 2012;7(8):e43474. doi: 10.1371/journal.pone.0043474. Epub 2012 Aug 27.
The tolerant mechanism of yeast to the combination of three inhibitors (furfural, phenol and acetic acid) was investigated using 2-DE combined with MALDI-TOF/TOF-MS. The stress response and detoxification related proteins (e.g., Ahp1p, Hsp26p) were expressed higher in the tolerant yeast than in the parental yeast. The expressions of most nitrogen metabolism related proteins (e.g. Gdh1p, Met1p) were higher in the parental yeast, indicating that the tolerant yeast decreases its nitrogen metabolism rate to reserve energy, and possesses high resistance to the stress of combined inhibitors. Furthermore, upon exposure to the inhibitors, the proteins related to protein folding, degradation and translation (e.g., Ssc1p, Ubp14p, Efb1p) were all significantly affected, and the oxidative stress related proteins (e.g., Ahp1p, Grx1p) were increased. Knockdown of genes related to the oxidative stress and unfolded protein response (Grx1, Gre2, Asc1) significantly decreased the tolerance of yeast to inhibitors, which further suggested that yeast responded to the inhibitors mainly by inducing unfolded protein response. This study reveals that increasing the detoxification and tolerating oxidative stress, and/or decreasing the nitrogen metabolism would be promising strategies in developing more tolerant strains to the multiple inhibitors in lignocellulose hydrolysates.
采用 2-DE 结合 MALDI-TOF/TOF-MS 技术研究了酵母耐受三种抑制剂(糠醛、苯酚和乙酸)组合的耐受机制。耐受酵母中应激反应和解毒相关蛋白(如 Ahp1p、Hsp26p)的表达高于亲本酵母。大多数氮代谢相关蛋白(如 Gdh1p、Met1p)在亲本酵母中的表达更高,表明耐受酵母降低氮代谢率以储备能量,对组合抑制剂的应激具有高抗性。此外,在受到抑制剂的影响后,与蛋白质折叠、降解和翻译相关的蛋白质(如 Ssc1p、Ubp14p、Efb1p)均受到显著影响,氧化应激相关蛋白(如 Ahp1p、Grx1p)增加。与氧化应激和未折叠蛋白反应相关的基因(Grx1、Gre2、Asc1)的敲低显著降低了酵母对抑制剂的耐受性,这进一步表明酵母主要通过诱导未折叠蛋白反应来应对抑制剂。本研究表明,提高解毒和耐受氧化应激的能力,和/或降低氮代谢可能是开发耐受木质纤维素水解物中多种抑制剂的更具耐受性菌株的有前途的策略。