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番茄果实ACC合成酶的纯化

胡建成, 华雪增, 刘愚, 梁厚果

兰州大学植物生理研究室. 甘肃,兰州730000；中科院上海植物生理研究所. 上海 200032

摘要

以伤诱导的番茄囊果皮为材料, 改进了ACC合成酶(EC4.4.1.14)纯化方法。经改进的5步纯化后, ACC合成酶被纯化7300倍, 比活性达到116 870U/mg, 回收率为7%, 以较少的步骤得到了较高的纯化倍数。纯化的ACC合成酶经SDS-PAGE和银染检验为一条带, 分子量为50kD, 等电点为pH6.5。图7表3参20

关键词： 番茄果实; ACC合成酶; 纯化

Studies on the purification of ACC synthase in tomato fruits

Hu Jiancheng, Hua Xuezeng, Liu Yu, Liang Houguo

Laboratory of Plant Physiology, Lanzhou University. Lanzhou 730000；China；Shanghai Institute of Plant Physiology, Chinese Academy of Sciences. Shanghai 200032

Abstract

The optimum time of incubation was found to be 10 h at 25℃ when the ACC synthase activity reached its maximum. Potassium phosphate buffer(PBP) and sodium phosphate buffer (PBS) showed the same effects on the extraction, but the reaction activity of the enzyme was twice as high in PBP as in PBS, indicating an activating effect of K~(+). PBP 100 mmol/L (pH 8.0, DTT 4 mmol/L, and PLP 5#mu#mol/L) was adopted as the extraction buffer. The use of polyethyleneglycol(PEG)6000(5%~13%)as the precipitant gave an ACC synthase activity twice as high as that of ammonium sulfate(40%~75%). The specific activity of ACC synthase was increased 7300 fold, reaching 116870 U/mg protein, and the yield was 7%. Purified ACC synthase showed a yellow strong band located at 50 kD after SDS-PAGE and silver staining. The amount of ACC synthase in tomato fruit pericarps induced by slicing was about 0.01% of the total soluble proteins.

Key words: Tomato fruit;ACC synthase;Purification

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引用本文：

胡建成, 华雪增, 刘愚, 梁厚果. 番茄果实ACC合成酶的纯化[J]. 生理学报 1995; 47 (1): .

Hu Jiancheng, Hua Xuezeng, Liu Yu, Liang Houguo. Studies on the purification of ACC synthase in tomato fruits. Acta Physiol Sin 1995; 47 (1): (in Chinese with English abstract).