Department of Chemical and Biomolecular Engineering, University of Houston , Houston, Texas 77204-4004, United States.
Department of Civil and Environmental Engineering, University of Houston , Houston, Texas 77204-4003, United States.
Langmuir. 2018 Jan 23;34(3):1133-1142. doi: 10.1021/acs.langmuir.7b02447. Epub 2017 Oct 17.
Adhesion of bacteria to interfaces is the first step in pathogenic infection, in biofilm formation, and in bioremediation of oil spills and other pollutants. Bacteria use a variety of surface structures to promote interfacial adhesion, with the level of expression of these structures varying in response to local conditions and environmental signals. Here, we investigated how overexpression of type 1 fimbriae, one such appendage, modifies the ability of Escherichia coli to adhere to solid substrates, via biofilm formation and yeast agglomeration, and to oil/water interfaces, via a microbial adhesion to hydrocarbon assay. A plasmid that enables inducible expression of E. coli MG1655 type 1 fimbriae was transformed into fimbriae-deficient mutant strain MG1655ΔfimA. The level of fimH gene expression in the engineered strain, measured using quantitative real-time PCR, could be tuned by changing the concentration of inducer isopropyl β-d-1-thiogalactopyranoside (IPTG), and was higher than that in strain MG1655. Increasing the degree of fimbriation only slightly modified the surface energy and zeta potential of the bacteria, but enhanced their ability to agglomerate yeast cells and to adhere to solid substrates (as measured by biofilm formation) and to oil/water interfaces. We anticipate that the tunable extent of fimbriation accessible with this engineered strain can be used to investigate how adhesin expression modifies the ability of bacteria to adhere to interfaces and to actively self-assemble there.
细菌与界面的黏附是致病感染、生物膜形成和生物修复溢油和其他污染物的第一步。细菌使用多种表面结构来促进界面黏附,这些结构的表达水平会根据局部条件和环境信号而变化。在这里,我们研究了通过生物膜形成和酵母聚集,以及通过微生物对烃类的附着测定,过度表达一种此类附属物——I 型菌毛,如何改变大肠杆菌黏附固体基质、油/水界面的能力。我们将能够诱导表达大肠杆菌 MG1655 型 I 菌毛的质粒转化到菌毛缺陷突变株 MG1655ΔfimA 中。通过改变诱导剂异丙基β-D-1-硫代半乳糖吡喃糖苷(IPTG)的浓度,我们可以测量工程菌株中 fimH 基因的表达水平,其表达水平高于 MG1655 菌株。增加菌毛化程度仅略微改变了细菌的表面能和zeta 电位,但增强了它们聚集酵母细胞以及黏附固体基质(通过生物膜形成测量)和油/水界面的能力。我们预计,通过这种工程菌株可以实现的可调节菌毛化程度,可用于研究黏附素表达如何改变细菌黏附界面和主动在界面上自我组装的能力。