State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen, 361005, PR China.
Microbes Environ. 2012;27(4):430-42. doi: 10.1264/jsme2.me12029. Epub 2012 Oct 5.
Roseobacter litoralis OCh149 is a type strain of aerobic anoxygenic phototrophic bacteria in marine Roseobacter clade. Its full genome has been sequenced; however, proteomic research, which will give deeper insights into the environmental stimuli on gene expression networks, has yet to be performed. In the present study, a proteomic approach was employed to analyze the status of R. litoralis OCh149 in carbon starvation during the stationary phase and its responses to a dark/light regimen (12 h:12 h) in both exponential and stationary phases. LC-MS/MS-based analysis of highly abundant proteins under carbon starvation revealed that proteins involved in transport, the transcription/translation process and carbohydrate metabolism were the major functional categories, while poly-β-hydroxyalkanoate (PHA), previously accumulated in cells, was remobilized after stress. Glucose, as the sole carbon source in the defined medium, was broken down by Entner-Doudoroff and reductive pentose phosphate (PP) pathways. Carbohydrate catabolism-related proteins were down-regulated under light regardless of the growth phase, probably due to inhibition of respiration by light. In contrast, responses of amino acid metabolisms to light regimen varied among different proteins during growth phases depending on cellular requirements for proliferation, growth or survival. Fluorescence induction and relaxation measurements suggested that functional absorption cross-sections of the photosynthetic complexes decreased during the dark period and always recovered to about the previous level during the light period. Although the photosynthetic genes in R. litoralis OCh149 are located on the plasmid, these data indicate the regulatory mechanism of photoheterotroph metabolism by both carbon and light availability.
海洋罗尔斯通氏菌(Roseobacter)中的好氧兼性光养菌 OCh149 是一株模式菌株。其全基因组序列已被测序;然而,对基因表达网络的环境刺激有更深入了解的蛋白质组学研究尚未进行。在本研究中,采用蛋白质组学方法分析了静止期碳饥饿条件下的 R. litoralis OCh149 状态及其在指数期和静止期的黑暗/光照(12 h:12 h)条件下的响应。基于 LC-MS/MS 的分析表明,在碳饥饿条件下,大量存在的蛋白质涉及运输、转录/翻译过程和碳水化合物代谢等主要功能类别,而先前在细胞中积累的聚-β-羟基烷酸(PHA)在应激后被重新利用。在定义的培养基中,葡萄糖作为唯一的碳源,通过 Entner-Doudoroff 和还原戊糖磷酸(PP)途径分解。无论生长阶段如何,光照都会下调碳水化合物代谢相关蛋白,这可能是由于光照抑制了呼吸作用。相比之下,氨基酸代谢对光照的响应在不同的蛋白质中在不同的生长阶段会有所不同,这取决于细胞增殖、生长或生存的需要。荧光诱导和弛豫测量表明,在黑暗期间,光合复合物的功能吸收截面减小,并且在光照期间总是恢复到大约之前的水平。尽管 R. litoralis OCh149 的光合基因位于质粒上,但这些数据表明了碳和光可用性对光异养代谢的调控机制。