Complementation of Sulfolobus solfataricus PBL2025 with an α-mannosidase: effects on surface attachment and biofilm formation
Complementation of Sulfolobus solfataricus PBL2025 with an α-mannosidase: effects on surface attachment and biofilm formation
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DOI：
10.1007/s00792-011-0411-2
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发表时间：
2012-01-01
期刊：
影响因子：
2.9
通讯作者：
Albers, S. -V.
中科院分区：
文献类型：
--
作者：
Koerdt, A.;Jachlewski, S.;Albers, S. -V.
与solfataricus P2相比，solfataricus突变体PBL2025缺失了50个基因（SSO3004-3050），包括可能参与糖降解或代谢的多种酶的编码基因。我们将缺失的两种蛋白，α -甘露糖苷酶（SSO3006, Ss α -man）和β -半乳糖苷酶LacS （SSO3019）补充到PBL2025中，并使用比较荧光显微镜和共聚焦激光扫描显微镜对重组菌株进行分析。我们证明Ss α -man互补菌株在细胞附着于玻璃和细胞在静态生物膜中的生长方面类似于S. solfataricus P2的行为。在表达Ss α -man时，重组菌株在表面附着和生物膜形成过程中葡萄糖和含甘露糖的细胞外聚合物（EPS）水平发生了变化，而LacS不发生变化。这些结果表明Ss α -man可能参与了s.s solfataricus细胞外糖基化蛋白聚糖树的去甘露糖基化和/或EPS组成的调节。另一方面，LacS在PBL2025中的表达降低了分离的总EPS的碳水化合物含量，这意味着在静态生物膜形成过程中调节了EPS的产生。这是第一批被确定在古细菌EPS形成中起作用的酶。
Compared to Sulfolobus solfataricus P2, the S. solfataricus mutant PBL2025 misses 50 genes (SSO3004-3050), including genes coding for a multitude of enzymes possibly involved in sugar degradation or metabolism. We complemented PBL2025 with two of the missing proteins, the alpha-mannosidase (SSO3006, Ss alpha-man) and the beta-galactosidase LacS (SSO3019), and performed comparative fluorescence microscopy and confocal laser scanning microscopy to analyze the recombinant strains. We demonstrated that the Ss alpha-man complemented strain resembled the S. solfataricus P2 behavior with respect to attachment of cells to glass and growth of cells in static biofilms. During expression of the Ss alpha-man, but not LacS, glucose and mannose-containing extracellular polymeric substance (EPS) levels changed in the recombinant strain during surface attachment and biofilm formation. These results suggest that the Ss alpha-man might be involved in the modulation of the EPS composition and/or in the de-mannosylation of the glycan tree, which is attached to extracellular glycosylated proteins in S. solfataricus. On the other hand, LacS expression in PBL2025 reduced the carbohydrate content of the isolated total EPS implying a role in the modulation of the produced EPS during static biofilm formation. These are the first enzymes identified as playing a role in archaeal EPS formation.