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|本期目录/Table of Contents|

巴西橡胶树胶乳磷脂酶A2的分离纯化及部分酶学性质鉴定

《广西植物》[ISSN:1000-3142/CN:45-1134/Q]

期数:
2010年06期
页码:
876-880
栏目:
植物生理学与分子生物学
出版日期:
2010-11-15

文章信息/Info

Title:
Purification and enzymatic characterization of phospholipase A2 from latex of Hevea brasiliensis
作者:
邓治 刘实忠 校现周*
中国热带农业科学院 橡胶研究所 农业部橡胶树生物学重点开放实验室,海南儋州 571737
Author(s):
DENG Zhi LIU Shi-Zhong XIAO Xian-Zhou*
Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences; Key Laboratory of Rubber Biology,Ministry of Agriculture,Danzhou,571737,China
关键词:
巴西橡胶树磷脂酶A2 酶的纯化酶的性质
Keywords:
Hevea brasiliensis phospholipase A2 purification of enzymes enzymological properties
分类号:
-
DOI:
-
文献标识码:
A
摘要:
通过丙酮沉淀、DEAE-纤维素离子交换柱层析和Sephadex G-100凝胶过滤柱层析等分离纯化技术,对巴西橡胶树(Hevea brasiliensis Muell. Arg.)胶乳C-乳清磷脂酶A2进行分离纯化。用SDS-PAGE测定其亚基的相对分子量。测定该酶最适温度和pH,动力学常数Km和Vmax。并测定Ca2+和La3+对酶活性的影响。结果显示该酶被纯化了49.47倍,产率为5.12%;SDS-PAGE检测为单一条带,其亚基相对分子量约43kDa。最适反应温度为37℃,最适反应pH为8.0,Km为0.44mmol ?L-1,Vmax为7.22μmol ?(mL?min)-1。最适Ca2+浓度为50?mol?L-1,稀土元素La3+离子对磷脂酶A2活性有抑制作用,但加入Ca2+后可缓解La3+对磷脂酶A2活性的抑制作用。胶乳C-乳清磷脂酶A2与其他植物磷脂酶A2在Ca2+的依赖性上存在差异。研究结果为今后探索橡胶树胶乳磷脂酶A2的催化机理、调节机理及生理功能等奠定了基础。
Abstract:
A phospholipase A2(PLA2)was purified from the latex C-serum of Hevea brasiliensis by acetone precipitation, DEAE-Cellulose ion exchange chromatography, Sephadex G-100 gel filtration. SDS-PAGE was used to determine the Mr of the subunit of PLA2. Then the Km, Vmax , the optimum temperature and pH of PLA2 was tested. The effect of Ca2+ and La3+ on PLA2 activity was assayed. A 49.47-fold purification was obtained with the recovery of 5.12% activity. The results showed that a single band on SDS-PAGE with a subunit Mr of 43kDa. Latex C-serum PLA2 had an optimum temperature at 37℃, and an optimum pH at 8.2. The Km was 0.44mmol ?L-1 . The Vmax was 7.22μmol?(mL?min) -1. The Ca2+ ion optimum concentration for activity was determined to be 50?mol?L-1. The activity of PLA2 was inhibited by La3+. The inhibited effect of La3+ was palliated after added calcium ion. There are differences of Ca2+ dependence between latex C-serum and other plant PLA2. The results provide a research basis for catalytic mechanism, regulation, and physiology function of PLA2 in latex.

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: 2011-01-28