Application of diethyl ethoxymethylenemalonate (DEEMM) derivatization for monitoring of lysine decarboxylase activity

作者：Kim Yong Hyun; Kim Hyun Joong; Shin Ji Hyun; Bhatia Shashi Kant; Seo Hyung Min; Kim Yun Gon; Lee Yoo Kyung; Yang Yung Hun^*; Park Kyungmoon

来源：Journal of Molecular Catalysis B: Enzymatic , 2015, 115: 151-154.

DOI：10.1016/j.molcatb.2015.01.018

• Highly sensitive method
• Diethyl ethoxymethylenemalonate
• Diamine analysis
• Cadevarine
• Corynebacterium glutamicum

Aliphatic diamines have been used as chemical reagents for the monomers of polyamides (i.e., Nylon 46, Nylon 510, Nylon 66, Nylon 610 and so on), and can be made from lysine, arginine or ornithine by the metabolism of microbes through decarboxylases. However, the conventional derivatization methods for the HPLC-based quantitative analysis of aliphatic diamines exhibit poor sensitivity and efficiency for the monitoring of enzyme activity. In this study, a chemical derivatization method using diethyl ethoxymethylenemalonate (DEEMM) was applied to monitor lysine decarboxylase activity for the production of cadaverine by measuring the levels of intracellular and secreted lysine and cadaverine. The calibration graphs for the determination of 2,4-diaminobutyrate, 2,6-diaminopimelic acid, ornithine, arginine, lysine, 1,3-diaminopropane, putrescine dihydrochloride, cadaverine, hexamethylenediamine, and 1,7-diaminoheptane were measured by the highly sensitive method using DEEMM, and a linear relationship was observed for each dimaine compound, with limits of detection up to 0.001 mM. Application of this method will be useful for the sensitive monitoring of lysine decarboxylase reactions through the detection of substrates containing amine groups and products with diamines using HPLC and a UV detector.

• 出版日期2015-5
• 单位中国极地研究中心