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鳄梨(Persea americana)亚细胞组分催化对氯-N-甲基苯胺的N-去甲基化反应。

N-demethylation of p-chloro-N-methylaniline catalyzed by subcellular fractions from the avocado pear (Persea americana).

作者信息

Dohn D R, Krieger R I

出版信息

Arch Biochem Biophys. 1984 Jun;231(2):416-23. doi: 10.1016/0003-9861(84)90405-3.

Abstract

Subcellular fractions from the avocado pear ( Persea americana) catalyzed formation of p-chloroaniline from p-chloro-N-methylaniline. Fractions prepared by centrifugation of avocado homogenates at 20, 000g for 20 min formed p-chloroaniline (2900 +/- 500 pmol min-1 mg protein-1) with an NADPH-generating system. p-Chloroaniline formation required reduced pyridine nucleotide (NADPH was 6-7 times more effective than NADH) and O2. N-Demethylation was inhibited by CO (55% inhibition at CO:O2 = 1) and was not inhibited by CN. Cytochrome P-450 was detected in the 20, 000g pellet at levels of 300-380 pmol/mg protein. This particulate preparation was also active in catalyzing the NADPH-dependent epoxidation of the chlorinated cyclodiene aldrin. Improvements to a colorimetric procedure for measuring p-chloroaniline increased the sensitivity of the procedure fourfold, and allowed use of samples containing high amounts of lipid. Avocado pear is suitable tissue for further studies on the oxidation of foreign compounds by higher plants.

摘要

鳄梨(Persea americana)的亚细胞组分催化了对氯-N-甲基苯胺生成对氯苯胺的反应。将鳄梨匀浆在20,000g下离心20分钟制备的组分,在有NADPH生成系统的情况下生成对氯苯胺(2900±500 pmol min⁻¹ mg蛋白质⁻¹)。对氯苯胺的生成需要还原型吡啶核苷酸(NADPH的效果比NADH高6 - 7倍)和O₂。N-去甲基化受到CO的抑制(在CO:O₂ = 1时抑制率为55%),且不受CN的抑制。在20,000g沉淀中检测到细胞色素P - 450,含量为300 - 380 pmol/mg蛋白质。这种颗粒制剂在催化氯化环二烯艾氏剂的NADPH依赖性环氧化反应中也具有活性。一种用于测量对氯苯胺的比色法经过改进后,该方法的灵敏度提高了四倍,并且可以使用含有大量脂质的样品。鳄梨是用于进一步研究高等植物对外源化合物氧化作用的合适组织。

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