Nagashima Kenji V P, Verméglio André, Fusada Naoki, Nagashima Sakiko, Shimada Keizo, Inoue Kazuhito
Research Institute for Photobiological Hydrogen Production, Kanagawa University, 2946 Tsuchiya, Hiratsuka, 259-1293, Kanagawa, Japan,
J Mol Evol. 2014 Aug;79(1-2):52-62. doi: 10.1007/s00239-014-9634-z. Epub 2014 Jul 31.
A mutant of the phototrophic species belonging to the β-proteobacteria, Rubrivivax gelatinosus, lacking the photosynthetic growth ability was constructed by the removal of genes coding for the L, M, and cytochrome subunits of the photosynthetic reaction center complex. The L, M, and cytochrome genes derived from five other species of proteobacteria, Acidiphilium rubrum, Allochromatium vinosum, Blastochloris viridis, Pheospirillum molischianum, and Roseateles depolymerans, and the L and M subunits from two other species, Rhodobacter sphaeroides and Rhodopseudomonas palustris, respectively, have been introduced into this mutant. Introduction of the genes from three of these seven species, Rte. depolymerans, Ach. vinosum, and Psp. molischianum, restored the photosynthetic growth ability of the mutant of Rvi. gelatinosus, although the growth rates were 1.5, 9.4, and 10.7 times slower, respectively, than that of the parent strain. Flash-induced kinetic measurements for the intact cells of these three mutants showed that the photo-oxidized cytochrome c bound to the introduced reaction center complex could be rereduced by electron donor proteins of Rvi. gelatinosus with a t1/2 of less than 10 ms. The reaction center core subunits of photosynthetic proteobacteria appear to be exchangeable if the sequence identities of the LM core subunits between donor and acceptor species are high enough, i.e., 70% or more.
通过去除编码光合反应中心复合物的L、M和细胞色素亚基的基因,构建了属于β-变形菌的光合营养物种胶质红环菌的一个缺乏光合生长能力的突变体。分别来自其他五种变形菌(嗜酸红菌、嗜酒全色菌、绿色绿硫菌、莫氏嗜盐菌和去聚玫瑰菌)的L、M和细胞色素基因,以及分别来自另外两种菌(球形红杆菌和沼泽红假单胞菌)的L和M亚基,已被导入该突变体。导入这七个物种中三个物种(去聚玫瑰菌、嗜酒全色菌和莫氏嗜盐菌)的基因后,恢复了胶质红环菌突变体的光合生长能力,尽管其生长速率分别比亲本菌株慢1.5倍、9.4倍和10.7倍。对这三个突变体的完整细胞进行闪光诱导动力学测量表明,与导入的反应中心复合物结合的光氧化细胞色素c可以被胶质红环菌的电子供体蛋白重新还原,半衰期小于10毫秒。如果供体和受体物种之间LM核心亚基的序列同一性足够高,即70%或更高,光合变形菌的反应中心核心亚基似乎是可交换的。