Novel DNA-Based Biocatalytic Nanomaterials

University Of Connecticut, Storrs CT

Investigators

Abstract

ID: MPS/DMR/BMAT(7623) 1005609 PI: Kumar, Challa ORG: University of Connecticut Title: Novel DNA-Based Biocatalytic Nanomaterials INTELLECTUAL MERIT: This proposal develops a strategy for attaching enzymes to solid substrates for use in various applications where their high chemo-, regio-, stereo- and chiral selectivities can be exploited. Because enzymes bound to solids are often less active than the corresponding free enzymes, the PI will initially examine the mechanism of enzyme binding to layered solids such as alpha-Zr(IV)phosphate/phosphonates (alpha-ZrRPs). The surface functions (R) of alpha-ZrRP will be varied systematically to control these interactions, and the PI will quantify the nature and magnitudes of the enthalpy and entropy contributions of specific processes that accompany the binding event, in order to elucidate the molecular signatures of enzyme-solid interactions. Using these insights, he will prepare rationally designed enzyme-DNA-ZrRP conjugates, under reagent-less conditions, via DNA-mediated self-assembly. The proposed approach consists of: (1) covalent attachment of single stranded DNA to specific ZrRP nanoparticles via a long tether, (2) linking of complementary single-stranded DNA to surface COOH groups of enzymes, and (3) hybridization of enzyme-DNA with DNA-ZrRP to produce enzyme-DNA-nanomaterials. Tethers will consist of oligoethyleneglycol (OEG) spacers as these are known to have little or no interaction with proteins or DNA. DNA hybridization is well known and it should result in enzyme- DNA-nanoparticles under regent-less conditions. DNA provides a robust, reversible, indirect link between the solid and the protein. DNA-labeled enzymes and nanoparticles will provide numerous opportunities to construct higher order assemblies for a variety of practical applications. BROADER IMPACTS: At the graduate level, the PI will continue to develop two new courses (Biological Chemistry I, and II, Chem360/361, 3 credits each) designed to train students in the application of chemical principles to important biological systems. Biological electron transfer, biomaterials, biocalorimetry, and DNA/protein structure are some of the topics contributed by the PI to these courses. The PI is developing a third graduate course in Technical Communications, Writing, and Ethics. At the undergraduate level, the PI will provide research training in biomaterials, protein-inorganic materials, and DNA-inorganic materials. Nearly 30 undergraduate students have been given such research experience in the PI's laboratory in the last 9 years as a part of the Chemistry Department Internship Program. Participating undergraduate students are often included as co-authors on resulting publications. At the high school level, the PI participates in the university-wide Mentor Connection program for outstanding high school students, sponsored by the University of Connecticut. He also hosts middle school students who participate in a day-long hands-on lab experience for students from throughout Connecticut.