Department of Soil, Plant and Food Science, Bari, University of Bari Aldo Moro, Bari, Italy.
Appl Environ Microbiol. 2013 Apr;79(8):2657-69. doi: 10.1128/AEM.03625-12. Epub 2013 Feb 8.
This study aimed at investigating the extracellular and cell wall-associated proteins (exoproteome) of Lactobacillus plantarum DC400 when cultivated on modified chemically defined medium (CDM) supplemented with the chemically synthesized pheromone plantaricin A (PlnA) or cocultured with L. plantarum DPPMA20 or Lactobacillus sanfranciscensis DPPMA174. Compared to monoculture, two-dimensional gel electrophoresis (2-DE) analysis showed that the exoproteome of L. plantarum DC400 was affected by PlnA and cocultivation with strains DPPMA20 and, especially, DPPMA174. The highest similarity of the 2-DE maps was found between DC400 cells cultivated in monoculture and in coculture with strain DPPMA20. Almost all extracellular proteins (22 spots) and cell wall-associated proteins (40 spots) which showed decreased or increased levels of synthesis during growth in CDM supplemented with PlnA and/or in coculture with strain DPPMA20 or DPPMA174 were identified. On the basis of the sequences in the Kyoto Encyclopedia of Genes and Genomes database, changes to the exoproteome concerned proteins involved in quorum sensing (QS), the transport system, stress response, carbohydrate metabolism and glycolysis, oxidation/reduction processes, the proteolytic system, amino acid metabolism, cell wall and catabolic processes, and cell shape, growth, and division. Cultivation with PlnA and cocultivation with strains DPPMA20 and, especially, DPMMA174 markedly increased the capacity of L. plantarum DC400 to form biofilms, to adhere to human Caco-2 cells, and to prevent the adhesion of potential intestinal pathogens. These phenotypic traits were in part related to oversynthesized moonlighting proteins (e.g., DnaK and GroEL, pyruvate kinase, enolase, and glyceraldehyde-3-phosphate dehydrogenase) in response to QS mechanisms and interaction with L. plantarum DPPMA20 and, especially, L. sanfranciscensis DPPMA174.
本研究旨在探究植物乳杆菌 DC400 在添加人工合成的细菌素 plantaricin A (PlnA) 的改良化学限定培养基 (CDM) 中培养或与植物乳杆菌 DPPMA20 或旧金山乳杆菌 DPPMA174 共培养时的胞外蛋白和细胞壁相关蛋白（外膜蛋白组）。与单一培养相比，二维凝胶电泳（2-DE）分析表明，PlnA 和与菌株 DPPMA20 及尤其是 DPPMA174 的共培养会影响植物乳杆菌 DC400 的外膜蛋白组。在 CDM 中培养的 DC400 细胞与在 CDM 中添加 PlnA 并与 DPPMA20 共培养的 2-DE 图谱的相似性最高。在添加 PlnA 和/或与 DPPMA20 或 DPPMA174 共培养时，所有表达量降低或升高的胞外蛋白（22 个斑点）和细胞壁相关蛋白（40 个斑点）都被鉴定出来。根据京都基因与基因组百科全书数据库中的序列，外膜蛋白组的变化涉及群体感应（QS）、转运系统、应激反应、碳水化合物代谢和糖酵解、氧化/还原过程、蛋白水解系统、氨基酸代谢、细胞壁和分解代谢过程以及细胞形态、生长和分裂相关的蛋白。添加 PlnA 和与 DPPMA20 及尤其是 DPMMA174 共培养显著提高了植物乳杆菌 DC400 形成生物膜、黏附于人 Caco-2 细胞以及阻止潜在肠道病原体黏附的能力。这些表型特征部分与 QS 机制相关的 moonlighting 蛋白（如 DnaK 和 GroEL、丙酮酸激酶、烯醇酶和甘油醛-3-磷酸脱氢酶）过度合成有关，还与与植物乳杆菌 DPPMA20 及尤其是旧金山乳杆菌 DPPMA174 的相互作用有关。
