Layer-by-Layer Encapsulation of Herbicide-Degrading Bacteria for Improved Surface Properties and Compatibility in Soils

Abstract

E. coli cells overexpressing the enzyme atrazine chlorohydrolase were coated using layer-by-layer self-assembly. The polymeric coating was designed to improve the surface properties of the cells and create positively charged, ecologically safe, bio-hybrid capsules that can efficiently degrade the herbicide atrazine in soils. The physio-chemical properties of the bacteria/polymer interface were studied as a function of the polymeric composition of the shell and its thickness. Characterization of cell viability, enzyme activity, morphology, and size of the bio-capsules was done using fluorescence spectroscopy, BET and zeta potential measurements and electron microscopy imaging. Out of several polyelectrolytes, the combination of polydiallyldimethylammonium chloride and polysodium 4-styrenesulfonate improved the surface properties and activity of the cells to the greatest extent. The resulting bio-hybrid capsules were stable, well-dispersed, with a net positive charge and a large surface area compared to the uncoated bacteria. These non-viable, bio-hybrid capsules also exhibited a kinetic advantage in comparison with uncoated cells. When added to soils, they exhibited continuous activity over a six-week period and atrazine concentrations declined by 84%. Thus, the concept of layer-by-layer coated bacteria is a promising avenue for the design of new and sustainable bioremediation and biocatalytic platforms.