Applications and Improvements of BioMEMS for Chitosan-Mediated Enzyme Assembly and Catalytic Activity

摘要

We report applications and improvements of bio- microelectricalmechanical systems (bioMEMS) in assembling catalytically active enzymes on readily addressable sites in microfluidic channels after complete fabrication and packaging of the bioMEMS device. We prefabricate an integrated bioMEMS device and exploit a non-permanent sealing and package design which supports programmable bio-component assembly at selected sites in the microfluidic environment. Chitosan's pH responsive and chemical reactive properties allow electrical signal-guided assembly of biomolecules onto selected inorganic area in the aqueous environment. Assembly of enzyme Pfs is based on covalent conjugation of Pfs to chitosan upon biochemical activation of a pentatyrosine pro-tag on Pfs followed by electrodeposition of the enzyme- chitosan conjugate onto readily addressable sites in microfluidic channels. The site-selectively assembled enzyme in the microfluidic channel retains stable catalytic activity for substantial cycles of enzymatic reactions, demonstrating our bioMEMS a powerful platform for biochemical and metabolic engineering applications. Currently, improvements of the bioMEMS are underway to optimize this microsystem for multi-step biochemical reaction applications.