One-Pot Production of Deoxyribonucleoside Triphosphates via Sequential Catalysis

描述（由申请人提供）：该SBIR I期项目旨在通过将所有三种酶与超顺磁性纳米晶体（纳米酶）结合，开发一种用于低成本商业生产脱氧核苷三磷酸的新型纳米技术。脱氧核苷三磷酸是DNA生物合成的前体，是生物医学和生物技术中不可缺少的化合物，市场规模达数十亿美元。这种新的纳米生物技术的目标是酶的再利用和生产工艺的简化，这使得由三个连续的酶促过程组成的一锅酶促生产方案成为可能。这种创新的纳米生物技术还大大减少了环境影响相比，传统的技术，用于制造脱氧核糖核苷三磷酸，通过排除多个中间纯化步骤，并简化了最后的纯化步骤，使用磁沉淀，除了重复使用的酶。这将减少缓冲试剂和水的消耗，减少废物产生，并消除有机溶剂的使用。该SBIR项目所需的纳米技术基础由高质量超顺磁性纳米晶体的专利大规模生产技术（由当前NSF SBIR第二阶段计划支持）和独特的配体化学技术，树枝状纳米晶体（与量子点相关的部分由NIH SBIR第二阶段计划支持）提供，NN-Labs拥有全部专利权。 预期效益和商业价值：SBIR计划的成功将大大提高脱氧核苷三磷酸的生产效率，降低生产成本。此外，这一新的方案将大大减少用于酶制备和中间体纯化的试剂和水的使用，并避免使用有毒的有机溶剂。脱氧核苷三磷酸是生物医学研究和生物技术的重要化合物。其应用范围包括DNA扩增、基因测序、基因组研究、分子诊断、法医检测和DNA芯片。脱氧核苷三磷酸及相关产品的市场需求是一个数十亿美元的业务部门。高价值和相对较小的生产量非常适合NN- Labs，一家拥有出色纳米技术基础的初创公司。与领先的生物试剂供应商的战略合作伙伴关系将极大地帮助NN-Labs进入这一市场。NN-Labs LLC作为该地区领先的创业公司，致力于在州政府和该州旗舰大学费耶特维尔阿肯色州大学的大力支持下，在这个传统的农村州发展知识型产业。拟议中的合作与大学通过一个研讨会将提供学生在阿肯色州大学独特的机会，暴露在高科技产业。

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-Enzymes). 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 of 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 for 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 (supported 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. Anticipated 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 sequencing, 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 productions 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 University through a subcontract will provide the students at the University of Arkansas unique opportunities to be exposed in high-tech industry.