Department of Biological Engineering, College of Engineering, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 05029, Republic of Korea.
Department of Biological and Chemical Engineering, Hongik University, Sejong Ro 2639, Jochiwon, Sejong City, 30016, Republic of Korea.
Anal Biochem. 2020 May 15;597:113688. doi: 10.1016/j.ab.2020.113688. Epub 2020 Mar 16.
Glutaric acid is a precursor of a plasticizer that can be used for the production of polyester amides, ester plasticizer, corrosion inhibitor, and others. Glutaric acid can be produced either via bioconversion or chemical synthesis, and some metabolites and intermediates are produced during the reaction. To ensure reaction efficiency, the substrates, intermediates, and products, especially in the bioconversion system, should be closely monitored. Until now, high performance liquid chromatography (HPLC) has generally been used to analyze the glutaric acid-related metabolites, although it demands separate time-consuming derivatization and non-derivatization analyses. To substitute for this unreasonable analytical method, we applied herein a gas chromatography - mass spectrometry (GC-MS) method with ethyl chloroformate (ECF) derivatization to simultaneously monitor the major metabolites. We determined the suitability of GC-MS analysis using defined concentrations of six metabolites (l-lysine, cadaverine, 5-aminovaleric acid, 2-oxoglutaric acid, glutamate, and glutaric acid) and their mass chromatograms, regression equations, regression coefficient values (R), dynamic ranges (mM), and retention times (RT). This method successfully monitored the production process in complex fermentation broth.
戊二酸是一种增塑剂的前体，可用于聚酯酰胺、酯类增塑剂、缓蚀剂等的生产。戊二酸可以通过生物转化或化学合成来生产，在反应过程中会产生一些代谢物和中间产物。为了确保反应效率，应密切监测底物、中间产物和产物，特别是在生物转化系统中。到目前为止，高效液相色谱法（HPLC）通常用于分析与戊二酸相关的代谢物，尽管它需要单独进行耗时的衍生化和非衍生化分析。为了替代这种不合理的分析方法，我们应用了一种带有乙基氯甲酸酯（ECF）衍生化的气相色谱-质谱法（GC-MS），同时监测主要代谢物。我们使用六种定义浓度的代谢物（赖氨酸、尸胺、5-氨基戊酸、2-氧代戊二酸、谷氨酸和戊二酸）及其质谱色谱图、回归方程、回归系数值（R）、动态范围（mM）和保留时间（RT）来确定 GC-MS 分析的适用性。该方法成功地监测了复杂发酵液中的生产过程。
