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烟草吡哆胺丙酮酸转氨酶的部分纯化与表征(PDF)

《广西植物》[ISSN:1000-3142/CN:45-1134/Q]

期数:
2013年01期
页码:
118
栏目:
植物化学与化学生理学
出版日期:
2013-01-25

文章信息/Info

Title:
Purification and enzymatic characterization of pyridoxaminepyruvate aminotransferase from the Tobacco
作者:
吴琼1 黄龙全1 张剑韵2*
1.安徽农业大学 茶与食品科技学院, 合肥 230036; 2.安徽农业大学 生命科学学院, 合肥 230036
Author(s):
WU Qiong1 HUANG LongQuan1 ZHANG JianYun2*
1.College of Tea & Food Sciences, Anhui Agricultural University, Hefei 230036, China; 2.College of Life Sciences, Anhui Agricultural University, Hefei 230036, China
关键词:
烟草 吡哆胺丙酮酸转氨酶 纯化 性质分析
Keywords:
tobacco pyridoxaminepyruvate aminotransferase purification properties
分类号:
Q945.14
DOI:
10.3969/j.issn.1000-3142.2013.01.021
文献标识码:
A
摘要:
通过冷冻干燥、DEAESepharose Fast Flow阴离子交换柱层析、Sephadex G100凝胶过滤柱层析,SP葡聚糖凝胶C25阳离子交换柱层析等分离纯化技术,对烟草吡哆胺丙酮酸转氨酶进行分离纯化。采用苯肼衍生化方法检测活性,并对其基本酶学性质进行分析。结果显示:该酶被纯化了92.34倍;最适温度为70 ℃,最适pH为9.0。在pH7.0~9.0内稳定且热稳定性较好,80 ℃保温3 h仍有51.55%的酶活力;在最适反应条件下,测得反应底物吡哆胺和丙酮酸的Km值分别为6.337 mmol?L1和0.867 mmol?L1。该结果为进一步研究烟草体内VB6代谢机制奠定了基础。
Abstract:
The pyridoxaminepyruvate aminotransferase was purified from the tobacco leaf by freezedrying,DEAE Sepharose Fast Flow ion exchange chromatography,Sephadex G100 gel filtration and SP Sephadex C25 ion exchange chromatography. The enzymatic activity and properties were investigated with phenyl hydrazine method. The results showed〖JP3〗 that 92.34fold purification was obtained;the enzyme had an optimum temperature at 70 ℃ and pH at 90,and it was stable at pH7.0-9.0;the enzyme had good thermal stability which remained about 51.55% of its activity after being treated at 80 ℃ for 3 h;under optimal conditions,the Km values for pyridoxamine and pyruvate were 6.337 mmol?L1 and 0.867 mmol?L1. The results provided basis for the metabolic mechanism of VB6 in tobacco plants.

参考文献/References

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备注/Memo

备注/Memo:
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更新日期/Last Update: 2013-01-25