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Near Infrared Spectroscopy As High-Throughput Technology for Screening of Xylose-Fermenting Recombinant Saccharomyces cerevisiae Strains

机译:近红外光谱作为高通量技术筛选木糖发酵啤酒酵母菌株的研究

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Recently, genetic engineering efforts have been made to develop recombinant Saccharomyces cerevisiae strains able to utilize xylose, an inexpensive and abundant carbon source. However, their construction and selection processes are limited by the speed and expenses of the existing testing methods, thus a rapid and equally precise method will significantly increase the number of tested strains. Here, near infrared (NIR) spectroscopy is proposed as a successful alternative method for screening recombinant xylose-fermenting S. cerevisiae. Supernatant samples of fermentation solutions from one diploid and three haploid recombinant strains were collected along the fermentation process. NIR spectra of the diluted supernatant provided effective differentiation of strains consistent with their phenotypic and genotypic features. This result could be used as a feedback for multicomponent analysis, in order to develop regression model for quantification of consumed glucose and xylose, produced ethanol, glycerol, and xylitol. Robust partial least-squares regression models with high prediction accuracy that are effective with any strain were achieved for all components when the modeling was performed with combined data of all strains, achieving 0.21-1.49 g/L of standard error of prediction with calibration, prediction, limit of detection and limit of quantification in the range of 1.0-4.5 and 3.0-13.4 g/L, respectively.
机译:最近,已经进行了基因工程研究以开发能够利用木糖的廉价酿酒酵母菌株,所述木糖是廉价且丰富的碳源。但是,它们的构建和选择过程受到现有测试方法的速度和费用的限制,因此,一种快速且同样精确的方法将显着增加测试菌株的数量。在这里,近红外(NIR)光谱被提议为筛选重组木糖发酵啤酒糖酵母的成功替代方法。在发酵过程中,收集了来自一个二倍体和三个单倍体重组菌株的发酵溶液上清液样品。稀释的上清液的NIR光谱可有效区分与其表型和基因型特征一致的菌株。该结果可用作多组分分析的反馈,以便开发用于定量分析消耗的葡萄糖和木糖,产生的乙醇,甘油和木糖醇的回归模型。当使用所有应变的组合数据进行建模时,对所有组件均获得了对任何组件均有效的,具有高预测精度的稳健的偏最小二乘回归模型,通过校准,预测获得了标准预测误差的0.21-1.49 g / L ,检测限和定量限分别在1.0-4.5和3.0-13.4 g / L的范围内。

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