Dispersing biofilms with engineered enzymatic bacteriophage

TLDR

Synthetic biology enables engineering of organisms to solve real‑world problems, and bacterial biofilms—resistant to antimicrobials and immune clearance—are a major clinical challenge. The study engineered bacteriophages to express a biofilm‑degrading enzyme during infection, aiming to simultaneously kill biofilm bacteria and degrade the extracellular matrix. The engineered phage delivers the enzyme to the biofilm matrix while infecting bacterial cells, creating a two‑pronged attack that targets both the cells and the matrix. This strategy removed approximately 99.997 % of biofilm cells—about 4.5 orders of magnitude—far exceeding the ~2‑order‑of‑magnitude reduction achieved by nonenzymatic phage.

Abstract

Synthetic biology involves the engineering of biological organisms by using modular and generalizable designs with the ultimate goal of developing useful solutions to real-world problems. One such problem involves bacterial biofilms, which are crucial in the pathogenesis of many clinically important infections and are difficult to eradicate because they exhibit resistance to antimicrobial treatments and removal by host immune systems. To address this issue, we engineered bacteriophage to express a biofilm-degrading enzyme during infection to simultaneously attack the bacterial cells in the biofilm and the biofilm matrix, which is composed of extracellular polymeric substances. We show that the efficacy of biofilm removal by this two-pronged enzymatic bacteriophage strategy is significantly greater than that of nonenzymatic bacteriophage treatment. Our engineered enzymatic phage substantially reduced bacterial biofilm cell counts by approximately 4.5 orders of magnitude ( approximately 99.997% removal), which was about two orders of magnitude better than that of nonenzymatic phage. This work demonstrates the feasibility and benefits of using engineered enzymatic bacteriophage to reduce bacterial biofilms and the applicability of synthetic biology to an important medical and industrial problem.