Department of Chemical and Biological Engineering , Tufts University , Medford , Massachusetts 02155 , United States.
ACS Synth Biol. 2020 Feb 21;9(2):356-367. doi: 10.1021/acssynbio.9b00356. Epub 2020 Jan 13.
Mucus in the gastrointestinal (GI) tract is the primary point-of-interaction between humans and their gut microbiota. This intimates that mucus not only ensures protection against endogenous and exogenous opportunists but also provisions for the human microbiota to reside and flourish. With the emergence of living therapeutics, engineered microbes can deliver and produce increasingly complex medicine, and controlling the mucoadhesive properties of different microbial chassis can dictate dose-response in a patient. Here we present a redesigned, , plate-based assay to measure the mucus adhesion of various probiotics. Cell-mucus interactions were isolated by immobilizing mucus to the plate surface. Binding parameters were derived for each probiotic strain by measuring cell adhesion over a wide range of cell concentrations, providing dose-dependent adhesion metrics. Surface proteins and cell components known to influence mucoadhesion were then heterologously expressed or altered in MG1363 and Nissle 1917 to control mucus-binding capacity, avidity, and cooperativity.
胃肠道（GI）中的黏液是人类与其肠道微生物群相互作用的主要切入点。这表明，黏液不仅能确保免受内源性和外源性机会致病菌的侵害，还能为人类微生物群提供栖息和繁殖的场所。随着活体治疗药物的出现，经过工程改造的微生物可以输送和生产越来越复杂的药物，并且控制不同微生物底盘的黏液附着特性可以决定患者的剂量反应。在这里，我们提出了一种经过重新设计的、基于平板的测定法，用于测量各种益生菌的黏液附着性。通过将黏液固定在平板表面，将细胞-黏液相互作用隔离开来。通过在广泛的细胞浓度范围内测量细胞黏附，为每个益生菌菌株推导出结合参数，从而提供了剂量依赖性黏附指标。然后，在 MG1363 和 Nissle 1917 中异源表达或改变已知影响黏液附着的表面蛋白和细胞成分，以控制黏液结合能力、亲合力和协同性。
