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Effects of acetic acid on the kinetics of xylose fermentation by an engineered, xylose-isomerase-based Saccharomyces cerevisiae strain

机译:乙酸对基于工程木糖异构酶的酿酒酵母菌株木糖发酵动力学的影响

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Acetic acid, an inhibitor released during hydrolysis of lignocellulosic feedstocks, has previously been shown to negatively affect the kinetics and stoichiometry of sugar fermentation by (engineered) Saccharomyces cerevisiae strains. This study investigates the effects of acetic acid on S. cerevisiae RWB 218, an engineered xylose-fermenting strain based on the Piromyces XylA (xylose isomerase) gene. Anaerobic batch cultures on synthetic medium supplemented with glucose-xylose mixtures were grown at pH 5 and 3.5, with and without addition of 3 g L(-1) acetic acid. In these cultures, consumption of the sugar mixtures followed a diauxic pattern. At pH 5, acetic acid addition caused increased glucose consumption rates, whereas specific xylose consumption rates were not significantly affected. In contrast, at pH 3.5 acetic acid had a strong and specific negative impact on xylose consumption rates, which, after glucose depletion, slowed down dramatically, leaving 50% of the xylose unused after 48 h of fermentation. Xylitol production was absent (<0.10 g L(-1)) in all cultures. Xylose fermentation in acetic -acid-stressed cultures at pH 3.5 could be restored by applying a continuous, limiting glucose feed, consistent with a key role of ATP regeneration in acetic acid tolerance.
机译:乙酸,一种在木质纤维素原料水解过程中释放的抑制剂,先前已显示出对(工程化)酿酒酵母菌株进行糖发酵的动力学和化学计量的不利影响。这项研究调查了乙酸对酿酒酵母RWB 218的影响,酿酒酵母RWB 218是一种基于Piromyces XylA(木糖异构酶)基因的工程木糖发酵菌株。在添加和不添加3 g L(-1)乙酸的条件下,在补充有葡萄糖-木糖混合物的合成培养基上的厌氧分批培养物在pH 5和3.5下生长。在这些文化中,糖混合物的消耗遵循双峰模式。在pH值为5时,添加乙酸会导致葡萄糖消耗率增加,而特定的木糖消耗率没有受到明显影响。相反,在pH 3.5时,乙酸对木糖的消耗速率具有强烈而特定的负面影响,葡萄糖消耗消失后,乙酸的木糖消耗速率急剧降低,发酵48小时后未使用的木糖占50%。在所有文化中都不存在木糖醇的生产(<0.10 g L(-1))。可以通过施加连续的有限葡萄糖进料来恢复乙酸在pH值为3.5的培养物中的木糖发酵,这与ATP再生在乙酸耐受性中的关键作用相一致。

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