A novel assay method for theanine synthetase activity by capillary electrophoresis |
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Authors: | Li Ping Wan Xiao-Chun Zhang Zheng-Zhu Li Jian Shen Zuo-Jun |
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Institution: | Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China. |
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Abstract: | The determination of theanine has been performed by micellar electrokinetic capillary chromatography (MECC) using 2,4-dinitrofluorobenzene (DNFB) as a derivative reagent. To achieve the separation, a fused-silica capillary column was used with a borax buffer at 0.03 mol/L pH 9.8 (containing Brij35 and isopropanol) at 17 degrees C with detection wave length at 360 nm. The factors affecting the efficiency of the sample separation were examined simultaneously. A 40-min reaction at 35 degrees C between l-glutamate and ethylamine (with Tris-HCl buffer, pH 7.5) was investigated using the theanine synthetase from budding tea seeds. A novel method for the analysis of theanine synthetase activity based on MECC was established. The method shows mean recovery ranged from 87.1 to 105.3% and linearity ranged from 0.2 to 5.0 mmol/L. |
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Keywords: | Micellar electrokinetic capillary chromatography Theanine Theanine synthetase Activity assay |
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