Laboratory of Microorganism Engineering, Beijing Institute of Biotechnology, 20 Dongdajie Street, Fengtai, Beijing, 100071, China.
World J Microbiol Biotechnol. 2013 Dec;29(12):2249-58. doi: 10.1007/s11274-013-1391-4. Epub 2013 Jun 5.
Methylovorus sp. MP688 is an aerobic bacterium that can grow on reduced C1 compounds such as methanol, being regarded as an attractive producer for many commercial materials including polysaccharides. The aim of the study was to learn more information about the biochemical and physiological functions of extracellular polysaccharides (EPS) produced by Methylovorus sp. MP688. Firstly, gene clusters involved in EPS synthesis were identified by whole genome sequence analysis. Then EPS produced by Methylovorus sp. MP688 were isolated and purified by centrifugation, precipitation and deproteinization. Purified EPS displayed antioxidant activity towards DPPH free radical, hydroxyl radical and superoxide anion radical. Glucose, galactose and mannose were identified to be main component monosaccharides in EPS. One mutant with defect in EPS production was obtained by knocking out epsA gene within EPS synthesis cluster. Strain with deletion of epsA exhibited compromised growth ability in the presence of oxidative stress due to the sharp reduction in EPS synthesis. Meanwhile, the intracellular antioxidant scavengers were activated to a higher level in order to counteract with the excess harmful radicals. In addition, EPS were assimilated by Methylovorus sp. MP688 to survive under disadvantage condition when the preferred carbon source was exhausted. It was reasonable to conclude that EPS produced by Methylovorus sp. MP688 contributed to oxidative defense and bacterial survival under adverse condition.
甲基球菌 MP688 是一种能够在甲醇等还原 C1 化合物上生长的需氧细菌,被认为是许多商业材料(包括多糖)的有吸引力的生产者。本研究的目的是更多地了解甲基球菌 MP688 产生的胞外多糖(EPS)的生化和生理功能。首先,通过全基因组序列分析鉴定了参与 EPS 合成的基因簇。然后通过离心、沉淀和脱蛋白法从甲基球菌 MP688 中分离和纯化 EPS。纯化的 EPS 对 DPPH 自由基、羟基自由基和超氧阴离子自由基具有抗氧化活性。鉴定出 EPS 的主要单糖组分为葡萄糖、半乳糖和甘露糖。通过敲除 EPS 合成簇中的 epsA 基因获得了一个 EPS 产量缺陷的突变体。由于 EPS 合成的急剧减少,缺失 epsA 的菌株在氧化应激存在下的生长能力受损。同时,为了对抗过量的有害自由基,细胞内抗氧化清除剂被激活到更高的水平。此外,当首选碳源耗尽时,EPS 被甲基球菌 MP688 同化以在不利条件下存活。因此,可以合理地得出结论,甲基球菌 MP688 产生的 EPS 有助于在不利条件下进行氧化防御和细菌存活。