Christiansen N, Ahring B K, Wohlfarth G, Diekert G
Department of Biotechnology, Technical University of Denmark, Lyngby.
FEBS Lett. 1998 Oct 2;436(2):159-62. doi: 10.1016/s0014-5793(98)01114-4.
The membrane-bound 3-chloro-4-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.
来自氯酚还原厌氧菌哈氏脱硫肠状菌的膜结合3-氯-4-羟基苯乙酸（Cl-OHPA）还原脱卤酶在去污剂CHAPS存在下纯化了11.3倍，达到表观均一性。纯化的脱卤酶以还原型甲基紫精作为电子供体，催化Cl-OHPA还原脱氯生成4-羟基苯乙酸，比活性为103.2 nkat/mg蛋白质。SDS-PAGE显示一条表观分子量为46.5 kDa的单一蛋白条带。该酶每摩尔亚基含有0.68±0.2摩尔类咕啉、12.0±0.7摩尔铁和13.0±0.7摩尔酸不稳定硫。测定了该酶的N端氨基酸序列，发现与基因库中任何蛋白质均无明显相似性。
