Kjaergaard K, Schembri M A, Ramos C, Molin S, Klemm P
Department of Microbiology, Technical University of Denmark, Lyngby.
Environ Microbiol. 2000 Dec;2(6):695-702. doi: 10.1046/j.1462-2920.2000.00152.x.
Antigen 43 (Ag43) is a surface-displayed autotransporter protein of Escherichia coli. By virtue of its self-association characteristics, this protein is able to mediate autoaggregation of E. coli cells in static cultures. Here, we show that Ag43 can be expressed in a functional form on the surface of Pseudomonas fluorescens. Ag43 expression dramatically enhances the biofilm-forming potential of both E. coli and P. fluorescens to abiotic surfaces in simple microtitre well assays and in flow chambers. Importantly, Ag43-expressing E. coli and P. fluorescens cells tagged with Gfp and Rfp were shown to form interwoven biofilms in flow chambers. The three-dimensional structures of the biofilms were analysed by laser-confocal microscopy. Heterogeneous expression of Ag43 induced interspecies cell-to-cell contact that generated multispecies biofilm formation. Our data indicate that this versatile molecular tool can be used for the rational design of multispecies biofilms. More specifically, this novel technology offers opportunities for the design of multispecies consortia in which the concerted action of several bacterial species is required, e.g. waste treatment and degradation of pollutants.
抗原43（Ag43）是大肠杆菌的一种表面展示自转运蛋白。凭借其自身缔合特性，该蛋白能够介导大肠杆菌细胞在静态培养物中的自动聚集。在此，我们表明Ag43能够以功能形式在荧光假单胞菌表面表达。在简单的微孔板试验和流动腔中，Ag43的表达显著增强了大肠杆菌和荧光假单胞菌在非生物表面形成生物膜的潜力。重要的是，用绿色荧光蛋白（Gfp）和红色荧光蛋白（Rfp）标记的表达Ag43的大肠杆菌和荧光假单胞菌细胞在流动腔中形成了交织的生物膜。通过激光共聚焦显微镜分析了生物膜的三维结构。Ag43的异源表达诱导了种间细胞间接触，从而产生了多物种生物膜的形成。我们的数据表明，这种多功能分子工具可用于合理设计多物种生物膜。更具体地说，这项新技术为设计需要几种细菌物种协同作用的多物种群落提供了机会，例如废物处理和污染物降解。
