SHMT酶促反应液中L-丝氨酸分离和有关组分分析

Separation of AminoL-Serine Acids and Assay of Some Components in SHMT catalyzed Reaction Solution

目的 :研究酶促反应液中L 丝氨酸的分离和反应液中甲醛、丝氨酸浓度的测定。方法 :采用 732树脂、71 7树脂离子交换柱层析从酶促反应液中分离甘氨酸、L 丝氨酸 ;采用铬变酸比色法测定甲醛和丝氨酸浓度。结果 :两种树脂柱层析均可不同程度地分离两种氨基酸 ,但 71 7树脂柱的分离效果更好 ,丝氨酸的收率可达 77%。结论 :利用 71 7树脂柱层析可从SHMT酶促反应液中成功地分离 L 丝氨 酸 ;铬变酸比色法可用于测定酶反应液中甲醛浓度。

Purpose:To study separation of L-ser from enzymatic reaction solution and determination of concentrations of bothL-ser and formaldehyde in the reaction solution. Methods:L-serine and glycine were separated from enzymeatic reaction solution by column chromatography on Resin 732 and Resin 717. The concentration ofL-serine and formaldehyde were determined by chromatic acid colorimetry. Results:The chromatography on both Resin 732 and Resin 717 could separate the two amino acids, but the separation effect of chromatography on Resin 717 was more satisfactory than that on Resin 732. The yield of L-serine was 77%. Conclusion:The satisfactory results of separation L-ser from SHMT catalyzed reaction soltion by chromatography on Resin 717 could be obtained under optimized condition. Colorimetry of chromatic acid may be used to determine the concentrations of L-serine and formaldehyde in the reaction solution.