Purification and characterization of the 3-chloro-4-hydroxy-phenylacetate reductive dehalogenase of Desulfitobacterium hafniense
Purification and characterization of the 3-chloro-4-hydroxy-phenylacetate reductive dehalogenase of Desulfitobacterium hafniense
复制标题
DOI：
10.1016/s0014-5793(98)01114-4
复制
发表时间：
1998-10-02
期刊：
影响因子：
3.5
通讯作者：
Diekert, G
中科院分区：
文献类型：
--
作者：
Christiansen, N;Ahring, BK;Diekert, G
从氯还原厌氧菌hafniense中提取的膜结合3-氯-3-羟基苯基乙酸（Cl-OHPA）还原脱卤酶在洗涤剂CHAPS存在下纯化了11.3倍，达到明显的均匀性。纯化后的脱卤酶以还原性紫甲基素为电子供体催化Cl-OHPA还原脱氯生成4-羟基苯基乙酸酯，比活性为103.2 nkat/mg蛋白，SDS-PAGE显示单个蛋白带的表观分子质量为46.5 kDa，酶每mol亚基含0.68 +/- 0.2 mol类碱、12.0 +/- 0.7 mol铁和13.0 +/- 0.7 mol酸不稳定硫。测定了该酶的n端氨基酸序列，并没有发现与基因库中存在的任何蛋白质有显著的相似性，(C) 1998 Federation of European Biochemical Societies。
The membrane-bound 3-chloro-3-hydroxyphenylacetate (Cl-OHPA) reductive dehalogenase from the chlorophenol-reducing anaerobe Desulfitobacterium hafniense was purified 11.3-fold to apparent homogeneity in the presence of the detergent CHAPS. The purified dehalogenase catalyzed the reductive dechlorination of Cl-OHPA to 4-hydroxyphenylacetate with reduced methyl viologen as the electron donor at a specific activity of 103.2 nkat/mg protein, SDS-PAGE revealed a single protein band with an apparent molecular mass of 46.5 kDa, The enzyme contained 0.68 +/- 0.2 mol corrinoid, 12.0 +/- 0.7 mol iron, and 13.0 +/- 0.7 mol acid-labile sulfur per mol subunit, The N-terminal amino acid sequence of the enzyme was determined and no significant similarity was found to any protein present in the gene bank, (C) 1998 Federation of European Biochemical Societies.