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Enhanced characterization of yeast hydrolysate combining acid digestion and 1D‐1H NMR targeted profiling
The Canadian Journal of Chemical Engineering ( IF 2.1 ) Pub Date : 2020-12-30 , DOI: 10.1002/cjce.24018
Marco Quattrociocchi 1 , Scott J. Boegel 1 , Marc G. Aucoin 1
Affiliation  

Yeast extract, or autolysate, is a required component for many cell‐culture media, but its exact constituents and benefits are unknown. Yeast extract contains a diverse assortment of metabolites, often present in complex forms (eg, polypeptides and polynucleotides). This study employs one‐dimensional proton nuclear magnetic resonance (1D‐1H NMR) spectroscopy to analyze free (ie, readily available) components present in commercially available yeast autolysate. The product is monitored while further subjected to acid hydrolysis, allowing for a more robust understanding of the exact components present, particularly those contained in complex forms. The amino acids and glucose compounds behaved as expected based on other acid hydrolysis studies, and were modelled similarly. Parameter estimation was in strong agreement with pre‐hydrolysis targeted 1D‐1H NMR profiling. This analysis was expanded to components not as thoroughly investigated and was especially applicable to nucleic compounds. Acid hydrolysis revealed that the yeast extract was approximately 5.4% nucleic material by weight, mostly composed of adenosine, and largely provided by RNA due to the presence of uracil and lack of thymidine. Choline‐containing compounds were also positively identified with an observable increase of free choline during hydrolysis. 1D‐1H NMR spectroscopy allowed for the simultaneous monitoring of a significant number of yeast‐extract metabolites, some of which were previously unreported. Utilizing 1D‐1H NMR spectroscopy in conjunction with acid hydrolysis led to a more complete view of the compounds present, and accounted for an additional 24% of the yeast‐extract mass.

中文翻译:

结合酸消解和1D-1H NMR靶向分析来增强酵母水解产物的表征

酵母提取物或自溶产物是许多细胞培养基的必需成分,但其​​确切成分和益处尚不清楚。酵母提取物包含各种各样的代谢产物,通常以复杂的形式(例如多肽和多核苷酸)存在。这项研究使用一维质子核磁共振波谱(1D-1H NMR)来分析市售酵母自溶物中存在的游离(即易于获得)组分。在进一步进行酸水解的同时对产物进行监控,从而可以更全面地了解存在的确切组分,尤其是复杂形式的组分。氨基酸和葡萄糖化合物的表现符合其他酸水解研究的预期,并且进行了类似的建模。参数估计与以水解前为目标的1D-1H NMR谱图非常吻合。该分析扩展到了尚未进行深入研究的组件,尤其适用于核酸化合物。酸水解显示,酵母提取物的核酸物质约占重量的5.4%,主要由腺苷组成,并且由于存在尿嘧啶和缺少胸苷,大部分由RNA提供。还可以肯定地鉴定出含胆碱的化合物在水解过程中游离胆碱的增加。1D-1H NMR光谱可同时监测大量酵母提取物代谢产物,其中一些以前未报道。将1D-1H NMR光谱与酸水解结合使用,可以更完整地了解存在的化合物,
更新日期:2021-03-10
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