Center for Nanomaterials and Devices, Kyoto Institute of Technology, Matsugasaki, Kyoto 606-8585, Japan.
Arch Microbiol. 2013 Feb;195(2):131-40. doi: 10.1007/s00203-012-0859-1. Epub 2012 Dec 21.
Scanning electron microscopy (SEM) shows remarkable morphological surface changes in Sphingopyxis sp. 113P3 cells grown in polyvinyl alcohol (PVA) but not in Luria-Bertani medium (LB) (Hu et al. in Arch Microbiol 188: 235-241, 2007). However, transmission electron microscopy showed no surface changes in PVA-grown cells and revealed the presence of polymer bodies in the periplasm of PVA-grown cells, which were not observed in LB-grown cells. The presence of polymer bodies was supported by low-vacuum SEM observation of PVA- and LB-grown cells of strain 113P3, and the presence of similar polymer bodies was also found when Sphingopyxis macrogoltabida 103 and S. terrae were grown in polyethylene glycol (PEG). The extraction of PVA and PEG from the periplasmic fraction of cells using a modified Anraku and Heppel method and their analysis by MALDI-TOF mass spectrometry strongly suggested that the polymer bodies are composed of PVA and PEG, respectively, in Sphingopyxis sp. 113P3 (PVA degrader) and Sphingopyxis macrogoltabida 103 or S. terrae (PEG degraders). PEG-grown S. macrogoltabida 103 and S. terrae showed higher transport of (14)C-PEG 4000 than LB-grown cells. Recombinant PegB (TonB-dependent receptor-like protein consisting of a barrel structure) interacted with PEG 200, 4000 and 20000, suggesting that the barrel protein in the outer membrane contributes to the transport of PEG into the periplasm.
扫描电子显微镜(SEM)显示,在聚乙烯醇(PVA)中生长的 Sphingopyxis sp. 113P3 细胞的表面形态发生了显著变化,但在 Luria-Bertani 培养基(LB)中则没有(Hu 等人,《Arch Microbiol》88:235-241, 2007)。然而,透射电子显微镜显示在 PVA 中生长的细胞表面没有变化,并揭示了在 PVA 中生长的细胞周质中存在聚合物体,而在 LB 中生长的细胞则没有观察到。低真空 SEM 观察到 PVA 和 LB 培养的 113P3 菌株的细胞,以及当 Sphingopyxis macrogoltabida 103 和 S. terrae 在聚乙二醇(PEG)中生长时发现类似的聚合物体,都支持了聚合物体的存在。使用改良的 Anraku 和 Heppel 方法从细胞周质部分提取 PVA 和 PEG,并通过 MALDI-TOF 质谱分析,强烈表明聚合物体分别由 Sphingopyxis sp. 113P3(PVA 降解物)和 Sphingopyxis macrogoltabida 103 或 S. terrae(PEG 降解物)中的 PVA 和 PEG 组成。在 PEG 中生长的 S. macrogoltabida 103 和 S. terrae 比在 LB 中生长的细胞对(14)C-PEG 4000 的转运更高。重组 PegB(由桶状结构组成的 TonB 依赖性受体样蛋白)与 PEG 200、4000 和 20000 相互作用,表明外膜中的桶状蛋白有助于 PEG 向周质的转运。