US EPA

8214343 

Journal Article 

Determination of serine: Glyoxylate aminotransferase activity by high performance liquid chromatography 

Zhu, Y; Lü, X; Wang, X; Wang, Y 

2003 

1 

Sepu / Chinese Journal of Chromatography
ISSN: 1000-8713
EISSN: 1872-2059 

21 

6 

584-586 

Chinese 

Serine: glyoxylate aminotransferase (SGAT, EC2. 6. 1. 45) is one of the key enzymes involved in the photorespiration pathway of plants. A method for the determination of SGAT activity by reversed-phase high performance liquid chromatography (HPLC) with 1-fluoro-2,4-dinitrobenzene (FDNB) pre-column derivatization is reported. The method has simple procedures and is accurate, rapid and efficient. The SGAT catalyzes the transfer of amino group from serine to glyxolate, producing glycine. Therefore, the increase in glycine contents in the reaction system can be analyzed by HPLC after derivatization and used to calculate the total activity of SGAT in the enzyme preparation. SGAT was extracted from plant tissues and an aliquot of the preparation was taken to incubate with L-serine, glyoxylate and pyridoxal 5′-phosphate at 30 °C for 60 min. The derivatization of the amino acids with FDNB was conducted at 60 °C for 60 min. After derivatization the sample was analyzed by HPLC with a C18 column. This method was applied to measure the SGAT activity in duckweed plants and reliable results were obtained. 

Activity determination; Glyoxylate aminotransferase; Reversed-phase high performance liquid chromatographyl serine