茶树鲜叶中维生素B6代谢酶PNPO的活性分析

doi:10.13305/j.cnki.jts.2010.04.007

茶叶科学 ›› 2010, Vol. 30 ›› Issue (4): 273-276.doi: 10.13305/j.cnki.jts.2010.04.007

茶树鲜叶中维生素B₆代谢酶PNPO的活性分析

李艳霞¹, 张剑韵², 黄龙全^1,*

1. 安徽农业大学茶与食品科技学院,安徽合肥 230036;
2. 安徽农业大学生命科学学院,安徽合肥 230036

收稿日期:2009-12-28 修回日期:2010-03-22 发布日期:2019-09-11
通讯作者: lqhuang218@yahoo.com.cn
作者简介:李艳霞（1983— ）,女,山东德州人,硕士研究生,主要从事茶叶生物化学研究。
基金资助:
安徽省教育厅自然科学重点科研项目（KJ2010A116）

Analysis on the Activity of PNPO of VB₆-metabolizing Enzymes from Tea Leaf

LI Yan-xia¹, ZHANG Jian-yun², HUANG Long-quan^1,*

1. Tea and Food Science College of AnHui Agricultural University, Hefei 230036, China;
2. Life Science College of Anhui Agricultural University, Hefei 230036, China

Received:2009-12-28 Revised:2010-03-22 Published:2019-09-11

摘要/Abstract

摘要： 采用液氮研磨、Tris-HCl+KPO₄缓冲液(pH8.5)浸提法和硫酸铵沉淀法从茶树鲜叶中提取磷酸吡哆醇氧化酶（PNPO）粗酶液,并通过苯肼法研究其酶学性质。结果表明,以磷酸吡哆胺（PMP）为底物,PNPO催化反应的最佳时间为30 min,最适温度为50℃,最适pH6.5。在pH6.0~9.0,温度10~40℃的条件下,此酶稳定。在最适条件下测得反应底物PMP的Km值为2.34 mmol/L。

关键词: 茶树, 磷酸吡哆醇氧化酶, 维生素B₆

Abstract: The crude extract of Pyridoxamine (Pyridoxine)-5′-phosphate Oxidase (PNPO) extracted from fresh tea leaves which were grinded by liquid nitrogen with Tris-HCL+ KPO buffer (pH8.5) and fractionated with solid ammonium sulfate, the enzymatic properties were investigated with phenyl hydrazine method. Results showed that when Pyridoxamine phosphate (PMP) as the substrate, the best time for the PNPO oxidation was 30 min, and the optimum pH was 6.5, and the optimum temperature was 50℃. When the value of pH was from 6.0 to 9.0 and temperature was from 10℃ to 40℃, the enzyme was stable. The Km_pmpof the reactive substrate PMP was 2.34 mmol/L under the optimum measure condition.

Key words: tea plant, pyridoxamine (Pyridoxine)-5′-phosphate oxidase, vitamin B₆

中图分类号:

李艳霞, 张剑韵, 黄龙全. 茶树鲜叶中维生素B₆代谢酶PNPO的活性分析[J]. 茶叶科学, 2010, 30(4): 273-276. doi: 10.13305/j.cnki.jts.2010.04.007.

LI Yan-xia, ZHANG Jian-yun, HUANG Long-quan. Analysis on the Activity of PNPO of VB₆-metabolizing Enzymes from Tea Leaf[J]. Journal of Tea Science, 2010, 30(4): 273-276. doi: 10.13305/j.cnki.jts.2010.04.007.

参考文献

[1] Roll L C, Mayodan S N.Vitamin b-6 and immune competence. Nutrition Reviews, 1993(51): 217-225.
[2] McCormick D B, Chen H. Update on introversions of vitamin b-6 with its coenzyme[J]. Journal of Nutrition, 1999, 129(2): 325-327.
[3] McCormick D B, Gregory M E, Snell E. Pyridoxal phosphor kinase I Assay, distribution, I Assay, distribution, purification, and properties[J]. Journal of Biological Chemistry, 1961(236): 2076-2084.
[4] Bilski P, Li M Y, Ehrenshaft M, et al. Vitamin B6 (pyridoxine) and its derivatives are efficient singlet oxygen quenchers and potential fungal antioxidants[J]. Photo chem Photobiology, 2000(71): 129-134.
[5] Shi H, Xing L, Stevenson B, et al. The Arabidopsis salt overly sensitive 4 mutants uncover a critical role for vitamin B6 in plant salt tolerance[J]. Plant Cell, 2002 (14): 575-588.
[6] Chen H, Xing L.Pyridoxine is required for postembryonic root development and tolerance to osmotic and oxidative stresses[J]. Plant J, 2005(44): 396-408.
[7] Eugenia Gonza´lez, David Danehower, Margaret E Daub. Vitamin Levels, Stress Response, Enzyme Activity,Gene Regulation of Arabidopsis Lines Mutant in the Pyridoxine/Pyridoxamine 5′-Phosphate Oxidase (PDX3) and the Pyridoxal Kinase (SOS4) Genes Involved in the Vitamin B₆ Salvage Pathway[J]. Plant Physiology, 2007(145): 985-996.
[8] Bradford M M.A rapid and sensitive method for the quantitation of microgram of protein utilizing the principle of protein-dye binding[J]. Anal Biochem, 1976(72): 248-254.
[9] Scopes R K.Protein purification: Principles and practice. 2nd Ed[M]. New York: Spring-Verlag New York Inc, 1988.
[10] HiroshiWada, Esmond E Snell. The Enzymatic Oxidation of Pyridoxine and Pyridoxamine Phosphates[J]. Journal of Biological Chemistry, 1961, 236(7): 2089-2095.
[11] Jung-DoChoi, Delores M, Bowers-Komro, et al. Kinetic properties of Pyridoxamine (pyridoxine)-5′-phosphate oxidase from rabbit Liver[J]. Journal of Biological Chemistry, 1983, 258(2): 840-845.

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茶树鲜叶中维生素B₆代谢酶PNPO的活性分析

Analysis on the Activity of PNPO of VB₆-metabolizing Enzymes from Tea Leaf

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