Nordling M, Olausson T, Lundberg L G
Department of Biochemistry and Biophysics, Chalmers University of Technology, Göteborg, Sweden.
FEBS Lett. 1990 Dec 10;276(1-2):98-102. doi: 10.1016/0014-5793(90)80517-m.
An expression vector designed for overexpression of plastocyanin in the periplasmic space of E. coli has been developed. The vector contains the signal peptide sequence of Pseudomonas aeruginosa azurin and the mature sequence of spinach plastocyanin. The precursor is efficiently translocated to the periplasmic space and correctly processed to mature plastocyanin. No detectable amount of plastocyanin was present in the cytoplasmic or in the membrane fraction. A large scale preparation of the recombinant plastocyanin in a 20 litre fermentor yielded approximately 30 mg of pure plastocyanin. The recombinant protein obtained from E. coli shows CD, EPR and optical properties identical to plastocyanin isolated from spinach.
已开发出一种用于在大肠杆菌周质空间中过表达质体蓝素的表达载体。该载体包含铜绿假单胞菌天青蛋白的信号肽序列和菠菜质体蓝素的成熟序列。前体可有效转运至周质空间并正确加工成成熟的质体蓝素。在细胞质或膜组分中未检测到质体蓝素。在20升发酵罐中大规模制备重组质体蓝素，可得到约30毫克的纯质体蓝素。从大肠杆菌获得的重组蛋白显示出与从菠菜中分离的质体蓝素相同的圆二色性（CD）、电子顺磁共振（EPR）和光学性质。
