Biofabrication is the assembly of biological components such as proteins, polysaccharides and nucleic acids using biological means or mimics thereof. Some of the products of biofabrication include sensor “chips” for biomedical applications like detection of cardiac diseases, tuberculosis, drug screening in blood etc. In these products, there is a need for quick and easy approach for assembly of multiple biological components.  Often, the interface material used in biofabrication determines the limitations and complexities of the biofabrication process. In this study, we used Tobacco Mosaic Virus- Virus like Particles (TMV-VLP’s)  as interface material to assemble a three enzyme pathway on chips to detect S-Adenosyl Methionine (SAM), a molecule implicated in neurological and psychiatric disorders including Alzheimer’s and depression.<br/>We created VLP’s wherein the TMV coat protein was engineered with N terminal Cysteines enabling self-assembly on gold surfaces and C terminal glutamines for bio-component assembly. We assembled biosynthetic enzymes onto VLP’s using a crosslinking cofactor, microbial transglutaminase that can covalently crosslink lysine and glutamine residues. We reengineered biosynthetic enzymes with lysine tags (7aa), enabling directed attachment to VLPs. In our system, we assembled a three-step enzymatic synthesis pathway onto the VLP’s using a modular protein assembly approach where we individually crosslinked each enzyme onto VLP’s and then assembled the VLP-enzyme complexes onto chips via user-specified stoichiometry to create a three enzyme pathway on gold chips.<br/>VLP’s as interface material provided significant advantages over earlier techniques in that they provide spatial control over the position of enzymes by enzymatic crosslinking. Rapid self-assembly of VLPs onto surfaces combined with precise control over the stoichiometries of enzymes assembled on the chips makes VLPs ideal for assembly of multi-component bio-devices on chips and microfluidic devices.<br/>

Fabrication of S-Adenosyl Methionine (SAM) biosensor via engineered virus like particles

253rd National Meeting (2017)

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