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Title:: Enzyme Properties Analysis of Glutamate-Pyruvate Transaminase in Soybean Hypocotyl

Author(s):: GUO Xu-shun; LIN Yu-tian; HE Jing-dong; LI Xin-hao; ZHANG Shang-hong; (1.School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China; 2.Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China)

Keywords:: Glycine max; Hypocotyl; Glutamate-pyruvate transaminase; Enzymatic properties; LC-MS/MS; Ninhydrin colorimetry

PACS:: -

DOI:: 10.11861/j.issn.1000-9841.2021.01.0082

Abstract:: Soybean (Glycine max) is an important nitrogen-fixing crop. It has strong amino acid metabolism and high expression of vigorous glutamate-pyruvate transaminase(GPT) during seed germination and early seedling growth. In this study, we identified the sequence of the GPT expressed in soybean hypocotyl through liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, and described its kinetic characterization to guide its application in genetic engineeringand agricultural production. The Km values of this enzyme for glutamate and pyruvate were 1.05 and 0.186 mmol?L-1,respectively, measured by ninhydrin colorimetry based on the chromogenic difference between glutamate and alanine. Co2+, Zn2+, Ca2+ and Mg2+ had promotion effect, while EDTA2-, urea, Mn2+ and Fe2+ had inhibitory effect on the enzyme. The enzyme was active in the temperature range between 10 and 60 ℃, with the optimum temperature of 35 ℃, and active in the pH range between 4.0 and 8.0, with the optimum pH7.0, measured by separating glutamate and alanine through paper chromatography and calculating the enzyme activity.

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Enzyme Properties Analysis of Glutamate-Pyruvate Transaminase in Soybean Hypocotyl(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

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