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High-Throughput Screening of Inhibitory Compounds on Growth and Ethanol Production of Saccharomyces cerevisiae

机译：高通量抑制化合物对酿酒酵母生长和乙醇生产的筛选

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摘要

A high-throughput screening experiment to establish the individual and mixture effects of various common inhibitors found in lignocellulosic hydrolysates on the growth kinetics and ethanol production of Saccharomyces cerevisiae was carried out. Fermentations were performed utilizing 96-well microtiter plates to allow for carrying out fermentations in parallel, around which a central composite design of experiments was used to select inhibitor concentrations in each well. The individual and interaction effects of six common inhibitors were quantified using response surface fits of the growth rate, as determined by optical absorbance measurements and final cell density. For growth rate, 4-hydroxy-methylbenzaldehyde (phenol aldehyde) was found to be the strongest inhibitor of growth rate over the concentration range studied, while m-cresol (phenolic) had the least effect on growth rate and the largest inhibitory effect on final cell density. Both positive and negative interactions between inhibitors were found to affect both growth rate and maximum cell density. For example, both furfural and guaiacol when combined with m-cresol were found to have a positive effect on cell growth (less inhibitory), while guaiacol and m-cresol had a negative interaction with 4-hydroxy-methylbenzaldehyde. At all conditions studied, S. cerevisiae produced identical or higher ethanol yields compared to the inhibitor-free control fermentation which was attributed to the effects of physiological stress cause by the some inhibitors. These results quantify both the interactions of various inhibitors as well as their individual effects in a rapid and easy-to-perform experiment which can be easily expanded to include further inhibitors. Such a design can also be used for rapid and efficient screening of different pretreatments and feedstocks in the emerging field of lignocellulosic biofuels.
机译：进行了高通量筛选实验，以建立木质纤维素水解物中发现的各种常见抑制剂对酿酒酵母的生长动力学和乙醇生产的单独作用和混合作用。利用96孔微量滴定板进行发酵，以允许平行进行发酵，在该孔周围，使用中央集中实验设计来选择每个孔中的抑制剂浓度。通过光学吸收率测量和最终细胞密度确定的生长速率的响应面拟合，对六种常见抑制剂的个体效应和相互作用效应进行了定量。就生长速率而言，发现4-羟基-甲基苯甲醛（酚醛）是在所研究浓度范围内最强的生长抑制剂，而间甲酚（酚醛）对生长速率的影响最小，对最终甲醛的抑制作用最大。细胞密度。发现抑制剂之间的正向和负向相互作用都影响生长速率和最大细胞密度。例如，糠醛和愈创木酚与间甲酚组合使用时，对细胞生长具有积极作用（抑制作用较小），而愈创木酚和间甲酚与4-羟基-甲基苯甲醛具有负相互作用。在所有研究的条件下，与无抑制剂的对照发酵相比，酿酒酵母产生相同或更高的乙醇产量，这归因于某些抑制剂引起的生理应激作用。这些结果在快速，易于执行的实验中量化了各种抑制剂的相互作用以及它们各自的作用，该实验可以轻松扩展为包括其他抑制剂。这种设计还可以用于快速有效地筛选新兴的木质纤维素生物燃料领域中的不同预处理和原料。

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  • 来源
    《Bioenergy research》 |2015年第1期|423-430|共8页
  • 作者

    Jeffery A. Wood; Valerie C. A. Orr; Luis Luque; Vivek Nagendra; Franco Berruti; Lars Rehmann;

  • 作者单位

    1.Department of Chemical and Biochemical Engineering University of Western Ontario London ON Canada;

    1.Department of Chemical and Biochemical Engineering University of Western Ontario London ON Canada;

    1.Department of Chemical and Biochemical Engineering University of Western Ontario London ON Canada;

    1.Department of Chemical and Biochemical Engineering University of Western Ontario London ON Canada;

    1.Department of Chemical and Biochemical Engineering University of Western Ontario London ON Canada;

    1.Department of Chemical and Biochemical Engineering University of Western Ontario London ON Canada;

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  • 正文语种 eng
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  • 关键词

    Inhibition High-throughput screening Yeast Biofuel Ethanol Growth kinetics;

    机译：抑制高通量筛选酵母生物燃料乙醇生长动力学;

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