†RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.
‡Graduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.
Environ Sci Technol. 2015 Jun 2;49(11):7056-62. doi: 10.1021/acs.est.5b00837. Epub 2015 May 14.
Planktonic metabolism plays crucial roles in Earth's elemental cycles. Chemical speciation as well as elemental stoichiometry is important for advancing our understanding of planktonic roles in biogeochemical cycles. In this study, a multicomponent solid-state nuclear magnetic resonance (NMR) approach is proposed for chemical speciation of cellular components, using several advanced NMR techniques. Measurements by ssNMR were performed on (13)C and (15)N-labeled Euglena gracilis, a flagellated protist. 3D dipolar-assisted rotational resonance, double-cross-polarization (1)H-(13)C correlation spectroscopy, and (1)H-(13)C solid-state heteronuclear single quantum correlation spectroscopy successively allowed characterization of cellular components. These techniques were then applied to E. gracilis cultured in high and low ammonium media to demonstrate the power of this method for profiling and comparing cellular components. Cellular NMR spectra indicated that ammonium induced both paramylon degradation and amination. Arginine was stored as a nitrogen reserve and ammonium replaced by arginine catabolism via the arginine dihydrolase pathway. (15)N and (31)P cellular ssNMR indicated arginine and polyphosphate accumulation in E. gracilis, respectively. This chemical speciation technique will contribute to environmental research by providing detailed information on environmental chemical properties.
浮游生物代谢在地球的元素循环中起着至关重要的作用。化学形态以及元素化学计量对于深入了解浮游生物在生物地球化学循环中的作用非常重要。在这项研究中,提出了一种多组分固态核磁共振(NMR)方法,用于细胞成分的化学形态分析,使用了几种先进的 NMR 技术。通过 ssNMR 对(13)C 和(15)N 标记的绿眼虫(一种鞭毛原生动物)进行了测量。3D 偶极辅助旋转共振、双交叉极化(1)H-(13)C 相关光谱和(1)H-(13)C 固态异核单量子相关光谱依次允许对细胞成分进行特征描述。然后将这些技术应用于在高铵和低铵培养基中培养的绿眼虫,以证明该方法用于分析和比较细胞成分的能力。细胞 NMR 谱表明,铵诱导了淀粉的降解和胺化。精氨酸被作为氮储备储存起来,并且通过精氨酸二氢酶途径,铵被精氨酸的分解代谢所取代。(15)N 和(31)P 细胞固态 NMR 分别表明绿眼虫中精氨酸和多磷酸盐的积累。这种化学形态分析技术将通过提供有关环境化学性质的详细信息,为环境研究做出贡献。