Gladstone Institutes, San Francisco, CA, USA.
Department of Biomedical Engineering, University of Calgary, Calgary, Canada.
Nat Commun. 2024 Nov 1;15(1):9443. doi: 10.1038/s41467-024-53546-1.
Spatial organization of microbes in biofilms enables crucial community function such as division of labor. However, quantitative understanding of such emergent community properties remains limited due to a scarcity of tools for patterning heterogeneous biofilms. Here we develop a synthetic optogenetic toolkit 'Multipattern Biofilm Lithography' for rational engineering and orthogonal patterning of multi-strain biofilms, inspired by successive adhesion and phenotypic differentiation in natural biofilms. We apply this toolkit to profile the growth dynamics of heterogeneous biofilm communities, and observe the emergence of spatially modulated commensal relationships due to shared antibiotic protection against the beta-lactam ampicillin. Supported by biophysical modeling, these results yield in-vivo measurements of key parameters, e.g., molecular beta-lactamase production per cell and length scale of antibiotic zone of protection. Our toolbox and associated findings provide quantitative insights into the spatial organization and distributed antibiotic protection within biofilms, with direct implications for future biofilm research and engineering.
生物膜中微生物的空间组织使分工等关键群落功能成为可能。然而，由于缺乏用于图案化异质生物膜的工具，对这种新兴群落特性的定量理解仍然有限。在这里，我们开发了一种合成的光遗传学工具包“多模式生物膜光刻”，用于合理工程和多菌株生物膜的正交图案化，这是受自然生物膜中连续粘附和表型分化的启发。我们将该工具包应用于异质生物膜群落的生长动力学分析，并观察到由于共享对β-内酰胺氨苄青霉素的抗生素保护而导致的空间调制共生关系的出现。通过生物物理建模的支持，这些结果提供了关键参数的体内测量值，例如，每个细胞的分子β-内酰胺酶产生量和抗生素保护带的长度尺度。我们的工具箱和相关发现为生物膜内的空间组织和分布式抗生素保护提供了定量见解，这对未来的生物膜研究和工程具有直接意义。
