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A rapid method for an offline glycerol determination during microbial fermentation

机译:微生物发酵过程中离线甘油快速测定的方法

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摘要

Background: The purpose of this work was to find a rapid method for glycerol detection during microbial fermentations. The method requirements were, first, to avoid sample pretreatment, and second, to measure glycerol precisely especially out of fermentation broth. Results: This was achieved by combining two reaction principles — the Malaprade reaction and the Hantzsch reaction. In the Malaprade reaction, glycerol is converted into formaldehyde. This forms a dye in the Hantzsch reaction after which adsorption is than detected. The subsequent assay was investigated with two different fermentation media, a chemically undefined and a chemically defined media, used for Pichia pastoris fermentation. In both media, as well as in real fermentation samples, glycerol content could be reproducibly detected with the method. Moreover, measurements were more precise than using a standard glycerol detection kit. Conclusions: With this rapid assay, glycerol could be detected easily in microbial fermentation broth. It is reliable over a wide concentration range including advantages such as an easy assay set-up, a short assay time and no sample pretreatment.
机译:背景:这项工作的目的是找到一种在微生物发酵过程中检测甘油的快速方法。方法要求首先是避免样品预处理,其次是精确测量甘油,尤其是发酵液中的甘油。结果:这是通过结合两种反应原理实现的:马拉普拉德反应和汉茨反应。在马拉普拉德反应中,甘油被转化为甲醛。这在Hantzsch反应中形成染料,然后检测到吸附。使用两种不同的发酵培养基(化学不确定的和化学确定的培养基)对巴斯德毕赤酵母发酵进行了后续测定。在两种培养基中以及在真实的发酵样品中,该方法都可以重现甘油含量。而且,测量比使用标准甘油检测试剂盒更精确。结论:通过这种快速测定,可以很容易地在微生物发酵液中检测到甘油。它在很宽的浓度范围内都是可靠的,包括诸如简便的测定设置,较短的测定时间以及无需样品预处理的优点。

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