Programmable Biomolecule Assembly and Activity in Prepackaged BioMEMS

Abstract

Antibiotic resistance is an increasing public health concern and few new drugs for bacterial pathogenesis have been obtained without addressing this resistance. Quorum sensing (QS) is a newly-discovered system mediated by extracellular chemical signals known as "autoinducers", which can coordinate population-scale changes in gene regulation when the number of cells reaches a "quorum" level. The capability to intercept and rewire the biosynthesis pathway of autoinduer-2 (AI-2), a universal chemical signaling molecule, opens the door to discover novel antimicrobial drugs that are able to bypass the antibiotic resistance. In this research, chitosan-mediated in situ biomolecule assembly has been demonstrated as a facile approach to direct the assembly of biological components into a prefabricated, systematically controlled bio-microelectromechanical system (bioMEMS). Our bioMEMS device enables post-fabricated, signal-guided assembly of labile biomolecules such as proteins and DNA onto localized inorganic surfaces inside microfluidic channels with spatial and temporal programmability. Particularly, the programmable assembly and enzymatic activity of the metabolic pathway enzyme Pfs, one of the two AI-2 synthases, have been demonstrated as an important step to reconstruct and interrogate the AI-2 synthesis pathway in the bioMEMS environment. Additionally, the bioMEMS has been optimized for studies of metabolic pathway enzymes by implementing a novel packaging technique and an experimental strategy to improve the signal-to-background ratio of the site-specific enzymatic reactions in the bioMEMS device. I envision that the demonstrated technologies represent a key step in progress toward a bioMEMS technology suitable to support metabolic engineering research and development.