Chávez S, Lucena J M, Reyes J C, Florencio F J, Candau P
Instituto de Bioquímica Vegetal y Fotosíntesis, Universidad de Sevilla-CSIC, 41092 Seville, Spain.
J Bacteriol. 1999 Feb;181(3):808-13. doi: 10.1128/JB.181.3.808-813.1999.
The unicellular cyanobacterium Synechocystis sp. strain PCC 6803 has two putative pathways for ammonium assimilation: the glutamine synthetase-glutamate synthase cycle, which is the main one and is finely regulated by the nitrogen source; and a high NADP-dependent glutamate dehydrogenase activity (NADP-GDH) whose contribution to glutamate synthesis is uncertain. To investigate the role of the latter, we used two engineered mutants, one lacking and another overproducing NADP-GDH. No major disturbances in the regulation of nitrogen-assimilating enzymes or in amino acids pools were detected in the null mutant, but phycobiline content, a sensitive indicator of the nutritional state of cyanobacterial cells, was significantly reduced, indicating that NADP-GDH plays an auxiliary role in ammonium assimilation. This effect was already prominent in the initial phase of growth, although differences in growth rate between the wild type and the mutants were observed at this stage only at low light intensities. However, the null mutant was unable to sustain growth at the late stage of the culture at the point when the wild type showed the maximum NADP-GDH activity, and died faster in ammonium-containing medium. Overexpression of NADP-GDH improved culture proliferation under moderate ammonium concentrations. Competition experiments between the wild type and the null mutant confirmed that the presence of NADP-GDH confers a selective advantage to Synechocystis sp. strain PCC 6803 in late stages of growth.
单细胞蓝藻聚球藻属6803菌株(Synechocystis sp. strain PCC 6803)有两条假定的铵同化途径:谷氨酰胺合成酶-谷氨酸合酶循环,这是主要途径,受氮源精细调控;以及一种高NADP依赖型谷氨酸脱氢酶活性(NADP-GDH),其对谷氨酸合成的贡献尚不确定。为了研究后者的作用,我们使用了两个工程突变体,一个缺失NADP-GDH,另一个过量表达NADP-GDH。在缺失突变体中未检测到氮同化酶调节或氨基酸库有重大干扰,但藻胆素含量(蓝藻细胞营养状态的敏感指标)显著降低,这表明NADP-GDH在铵同化中起辅助作用。这种效应在生长初期就已很明显,尽管在这个阶段仅在低光照强度下观察到野生型和突变体之间的生长速率差异。然而,在培养后期,当野生型显示出最大NADP-GDH活性时,缺失突变体无法维持生长,并且在含铵培养基中死亡更快。NADP-GDH的过表达在中等铵浓度下改善了培养物增殖。野生型和缺失突变体之间的竞争实验证实,NADP-GDH的存在赋予聚球藻属6803菌株在生长后期的选择性优势。