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Process intensification through microbial strain evolution: mixed glucose-xylose fermentation in wheat straw hydrolyzates by three generations of recombinant Saccharomyces cerevisiae

机译:通过微生物菌株的进化强化过程:三代重组酿酒酵母在麦秸水解物中混合葡萄糖-木糖发酵

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摘要

BackgroundLignocellulose hydrolyzates present difficult substrates for ethanol production by the most commonly applied microorganism in the fermentation industries, Saccharomyces cerevisiae. High resistance towards inhibitors released during pretreatment and hydrolysis of the feedstock as well as efficient utilization of hexose and pentose sugars constitute major challenges in the development of S. cerevisiae strains for biomass-to-ethanol processes. Metabolic engineering and laboratory evolution are applied, alone and in combination, to adduce desired strain properties. However, physiological requirements for robust performance of S. cerevisiae in the conversion of lignocellulose hydrolyzates are not well understood. The herein presented S. cerevisiae strains IBB10A02 and IBB10B05 are descendants of strain BP10001, which was previously derived from the widely used strain CEN.PK 113-5D through introduction of a largely redox-neutral oxidoreductive xylose assimilation pathway. The IBB strains were obtained by a two-step laboratory evolution that selected for fast xylose fermentation in combination with anaerobic growth before (IBB10A02) and after adaption in repeated xylose fermentations (IBB10B05). Enzymatic hydrolyzates were prepared from up to 15% dry mass pretreated (steam explosion) wheat straw and contained glucose and xylose in a mass ratio of approximately 2.
机译:背景技术木质纤维素水解产物是发酵工业中最常用的微生物酿酒酵母(Saccharomyces cerevisiae)产生乙醇的困难底物。对原料的预处理和水解过程中释放的抑制剂的高抗性以及己糖和戊糖的有效利用,构成了用于生物质制乙醇工艺的酿酒酵母菌株开发的主要挑战。单独或组合应用代谢工程和实验室改进,以产生所需的应变特性。然而,对于酿酒酵母在木质纤维素水解产物的转化中的稳健性能的生理要求还没有被很好地理解。本文呈现的酿酒酵母菌株IBB10A02和IBB10B05是菌株BP10001的后代,其先前通过引入大量的氧化还原-中性氧化还原木糖同化途径衍生自广泛使用的菌株CEN.PK 113-5D。 IBB菌株是通过两步实验室演变而获得的,该过程选择了快速木糖发酵结合厌氧生长,然后再适应木糖发酵(IBB10B05),然后进行厌氧生长。酶水解物由高达15%干燥质量的预处理(蒸汽爆炸)小麦秸秆制得,并包含质量比约为2的葡萄糖和木糖。

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