Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, ND 58108-6050, USA.
Arch Microbiol. 2010 Sep;192(9):715-28. doi: 10.1007/s00203-010-0599-z. Epub 2010 Jun 18.
Biofilms are communities of bacteria whose formation on surfaces requires a large portion of the bacteria's transcriptional network. To identify environmental conditions and transcriptional regulators that contribute to sensing these conditions, we used a high-throughput approach to monitor biofilm biomass produced by an isogenic set of Escherichia coli K-12 strains grown under combinations of environmental conditions. Of the environmental combinations, growth in tryptic soy broth at 37 degrees C supported the most biofilm production. To analyze the complex relationships between the diverse cell-surface organelles, transcriptional regulators, and metabolic enzymes represented by the tested mutant set, we used a novel vector-item pattern-mining algorithm. The algorithm related biofilm amounts to the functional annotations of each mutated protein. The pattern with the best statistical significance was the gene ontology 'pyruvate catabolic process,' which is associated with enzymes of acetate metabolism. Phenotype microarray experiments illustrated that carbon sources that are metabolized to acetyl-coenzyme A, acetyl phosphate, and acetate are particularly supportive of biofilm formation. Scanning electron microscopy revealed structural differences between mutants that lack acetate metabolism enzymes and their parent and confirmed the quantitative differences. We conclude that acetate metabolism functions as a metabolic sensor, transmitting changes in environmental conditions to biofilm biomass and structure.
生物膜是由细菌组成的群落,其表面形成需要细菌转录网络的很大一部分。为了确定有助于感知这些条件的环境条件和转录调节剂,我们使用高通量方法来监测在环境条件组合下生长的一组同源大肠杆菌 K-12 菌株产生的生物膜生物量。在 37 摄氏度的胰蛋白酶大豆肉汤中生长的环境组合支持最多的生物膜生产。为了分析由测试的突变体集代表的多样化细胞表面器官、转录调节剂和代谢酶之间的复杂关系,我们使用了一种新的向量-项目模式挖掘算法。该算法将生物膜数量与每个突变蛋白的功能注释相关联。具有最佳统计显著性的模式是与乙酸代谢酶相关的基因本体论“丙酮酸分解代谢过程”。表型微阵列实验说明了代谢为乙酰辅酶 A、乙酰磷酸和乙酸的碳源特别有助于生物膜的形成。扫描电子显微镜揭示了缺乏乙酸代谢酶的突变体与其亲本之间的结构差异,并证实了定量差异。我们得出结论,乙酸代谢作为代谢传感器,将环境条件的变化传递给生物膜生物量和结构。