One-Pot Production of Deoxyribonucleoside Triphosphates via Sequential Catalysis

DESCRIPTION (provided by applicant): This SBIR Phase I project intends to develop a novel nanotechnology for low-cost commercial production of deoxyribonucleoside triphosphates by conjugating all three enzymes with superparamagnetic nanocrystals (Nano-Enzy mes). Deoxiribonucleoside triphosphates, with a billion-dollar market, are the precursors for DNA biosynthesis and the indispensable compounds for biomedicine and biotechnology. This new nano- biotechnology targets the reuse of enzymes and simplification o f the production processes, which enables a one-pot enzymatic production protocol consisting of three sequential enzymatic processes. This innovative nano-biotechnology also greatly reduces environmental impact in comparison to the traditional technology f or making deoxyribonucleoside triphosphates by excluding multiple intermediate purification steps and simplifying the last purification step using magnetic precipitation, in addition to reuse of the enzymes. This shall decrease the consumption of buffering reagents and water, reduce waste production, and eliminate the use of organic solvents. The nanotechnology basis needed for this SBIR project is provided by the patented large- scale production technology for high quality superparamagnetic nanocrystals (s upported by a current NSF SBIR Phase II program) and a unique ligand chemistry technology, dendron-nanocrystals (the part related to quantum dots being supported by a NIH SBIR Phase II Program), to which NN-Labs holds full patent rights. Anticipate d Benefits and Commercial Value: Success of this SBIR program will greatly increase deoxyribonucleoside triphosphates production efficiency and lower the production cost. In addition, this new protocol will greatly reduce the use of reagents and water for enzyme preparation and intermediate purification and avoid the use of toxic organic solvents. Deoxyribonucleoside triphosphates are essential compounds for biomedical research and biotechnology. Their applications range from DNA amplification, gene sequenc ing, genomic research, molecular diagnostics, forensic detection, and DNA chips. The market demand for deoxyribonucleoside triphosphates and related products is a multi-billion-dollar business sector. The high value and relatively small volume of productio ns are well suited for NN- Labs, a startup company with an outstanding nanotechnology base. The strategic partnership with leading bio-reagents supplier will greatly help NN-Labs to enter this market. NN-Labs LLC, as a leading startup company in the area, is committed to develop knowledge-based industry in this traditionally rural state with the strong support of the State government and the flagship university in the state, the University of Arkansas at Fayetteville. The proposed collaboration with the Uni versity through a subcontract will provide the students at the University of Arkansas unique opportunities to be exposed in high-tech industry.