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Adaptation to low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and growth of Saccharomyces cerevisiae

机译:适应低pH值和木质纤维素抑制剂导致乙醇发酵和酿酒酵母的生长

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摘要

Lignocellulosic bioethanol from renewable feedstocks using Saccharomyces cerevisiae is a promising alternative to fossil fuels owing to environmental challenges. S. cerevisiae is frequently challenged by bacterial contamination and a combination of lignocellulosic inhibitors formed during the pre-treatment, in terms of growth, ethanol yield and productivity. We investigated the phenotypic robustness of a brewing yeast strain TMB3500 and its ability to adapt to low pH thereby preventing bacterial contamination along with lignocellulosic inhibitors by short-term adaptation and adaptive lab evolution (ALE). The short-term adaptation strategy was used to investigate the inherent ability of strain TMB3500 to activate a robust phenotype involving pre-culturing yeast cells in defined medium with lignocellulosic inhibitors at pH 5.0 until late exponential phase prior to inoculating them in defined media with the same inhibitor cocktail at pH 3.7. Adapted cells were able to grow aerobically, ferment anaerobically (glucose exhaustion by 19 ± 5 h to yield 0.45 ± 0.01 g ethanol g glucose−1) and portray significant detoxification of inhibitors at pH 3.7, when compared to non-adapted cells. ALE was performed to investigate whether a stable strain could be developed to grow and ferment at low pH with lignocellulosic inhibitors in a continuous suspension culture. Though a robust population was obtained after 3600 h with an ability to grow and ferment at pH 3.7 with inhibitors, inhibitor robustness was not stable as indicated by the characterisation of the evolved culture possibly due to phenotypic plasticity. With further research, this short-term adaptation and low pH strategy could be successfully applied in lignocellulosic ethanol plants to prevent bacterial contamination.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0234-8) contains supplementary material, which is available to authorized users.
机译:由于环境挑战,使用酿酒酵母(Saccharomyces cerevisiae)来自可再生原料的木质纤维素生物乙醇是化石燃料的有前途的替代品。在生长,乙醇产量和生产率方面,啤酒酵母经常受到细菌污染和在预处理期间形成的木质纤维素抑制剂的组合的挑战。我们研究了酿造酵母菌株TMB3500的表型稳健性及其适应低pH的能力,从而通过短期适应和适应性实验室进化(ALE)防止细菌与木质纤维素抑制剂一起污染。短期适应策略用于研究菌株TMB3500激活强大表型的内在能力,涉及在具有pH 5.0的木质纤维素抑制剂的特定培养基中预培养酵母细胞直至指数后期,然后再在相同的培养基中接种酵母pH 3.7的抑制剂混合物。适应的细胞能够有氧生长,厌氧发酵(葡萄糖耗竭19±5小时以产生0.45±0.01 g乙醇g葡萄糖 -1 ),与pH 3.7相比,抑制剂具有明显的解毒作用。非适应性细胞。进行ALE以研究是否可以在连续悬浮培养中用木质纤维素抑制剂开发出稳定的菌株以在低pH下生长和发酵。尽管在3600小时后获得了强大的种群,并且能够在pH 3.7的条件下与抑制剂一起生长和发酵,但正如表述可培养物的特性所表明的那样,抑制剂的耐受性并不稳定,这可能是由于表型可塑性所致。通过进一步的研究,这种短期适应性和低pH的策略可以成功地应用于木质纤维素乙醇工厂中以防止细菌污染。电子补充材料本文的在线版本(doi:10.1186 / s13568-016-0234-8)包含补充材料,可供授权用户使用。

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