Putative Alginate Assimilation Process of the Marine Bacterium Saccharophagus degradans 2-40 Based on Quantitative Proteomic Analysis
Putative Alginate Assimilation Process of the Marine Bacterium Saccharophagus degradans 2-40 Based on Quantitative Proteomic Analysis
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DOI：
10.1007/s10126-015-9667-3
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发表时间：
2015-10
影响因子：
3
通讯作者：
T. Takagi;Hironobu Morisaka;Shunsuke Aburaya;Yohei Tatsukami;K. Kuroda;M. Ueda
中科院分区：
文献类型：
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作者：
T. Takagi;Hironobu Morisaka;Shunsuke Aburaya;Yohei Tatsukami;K. Kuroda;M. Ueda
利用蛋白质组学定量分析技术研究了葡萄糖、果胶和海藻酸钠作为碳源培养的降解食蟹猴的同化过程。液相色谱-串联质谱法，采用我们独特的，长的整体硅胶毛细管柱。在试图选择与藻酸盐同化相关的候选蛋白质时，通过定量蛋白质组学数据的统计分析鉴定了23种藻酸盐特异性蛋白质的产生。在此基础上，我们提出S. degradans是一个藻酸盐特异性基因簇，可有效利用藻酸盐。S.降解产物由藻酸盐裂解酶、与碳水化合物代谢相关的酶、膜转运蛋白和转录因子组成。其中，在藻酸盐特异性簇中注释的短链脱氢酶/还原酶Sde_3281显示出4-脱氧-1-异丙基-5-己糖基葡糖醛酸还原酶（DehR）活性。此外，我们发现了两个不同的基因（Sde_3280和Sde_0939）编码的2-酮-3-脱氧-d-葡萄糖酸（KDG）激酶（KdgK）代谢的KDG来源于藻酸盐和果胶在降解链霉菌。S.降解物使用Sde_3280磷酸化来源于藻酸盐的KDG，使用Sde_0939磷酸化来源于果胶的KDG。KdgKs的独特选择为阐明howS提供了重要线索。降解物识别和加工多糖。
Quantitative proteomic analysis was conducted to assess the assimilation processes ofSaccharophagus degradanscultured with glucose, pectin, and alginate as carbon sources. A liquid chromatography-tandem mass spectrometry approach was used, employing our unique, long monolithic silica capillary column. In an attempt to select candidate proteins that correlated to alginate assimilation, the production of 23 alginate-specific proteins was identified by statistical analyses of the quantitative proteomic data. Based on the analysis, we propose thatS. degradanshas an alginate-specific gene cluster for efficient alginate utilization. The alginate-specific proteins ofS. degradanswere comprised of alginate lyases, enzymes related to carbohydrate metabolism, membrane transporters, and transcription factors. Among them, the short-chain dehydrogenase/reductase Sde_3281 annotated in the alginate-specific cluster showed 4-deoxy-l-erythro-5-hexoseulose uronic acid reductase (DehR) activity. Furthermore, we found two different genes (Sde_3280andSde_0939) encoding 2-keto-3-deoxy-d-gluconic acid (KDG) kinases (KdgK) that metabolize the KDG derived from alginate and pectin inS. degradans. S. degradansused Sde_3280 to phosphorylate the KDG derived from alginate and Sde_0939 to phosphorylate the KDG derived from pectin. The distinct selection of KdgKs provides an important clue toward the elucidation of howS. degradansrecognizes and processes polysaccharides.