One-Pot Production of Deoxyribonucleoside Triphosphates via Sequential Catalysis

描述(申请人提供)：这个SBIR第一阶段项目旨在开发一种新的纳米技术，通过将所有三种酶与超顺磁性纳米晶体(纳米酶)连接起来，低成本地商业生产脱氧核糖核苷三磷酸。脱氧核糖核苷三磷酸是DNA生物合成的前体，也是生物医学和生物技术不可缺少的化合物，市场价值高达数十亿美元。这种新的纳米生物技术的目标是酶的重复使用和生产过程的简化，这使得一锅酶生产方案能够由三个连续的酶过程组成。与传统的制备脱氧核糖核苷三磷酸的技术相比，这种创新的纳米生物技术还大大减少了对环境的影响，因为它省去了多个中间纯化步骤，并简化了使用磁性沉淀法的最后一个纯化步骤，此外还可以重复使用酶。这将减少缓冲剂和水的消耗，减少废物产生，并消除有机溶剂的使用。这个SBIR项目所需的纳米技术基础是高质量超顺磁性纳米晶体的专利大规模生产技术(由当前的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)，NN-Labs拥有全部专利权。 预期效益和商业价值：SBIR项目的成功将极大地提高脱氧核糖核苷三磷酸的生产效率，降低生产成本。此外，这一新方案将大大减少酶制备和中间体纯化所使用的试剂和水，并避免使用有毒的有机溶剂。脱氧核糖核苷三磷酸是生物医学研究和生物技术中必不可少的化合物。它们的应用范围包括DNA扩增、基因测序、基因组研究、分子诊断、法医检测和DNA芯片。市场对脱氧核糖核苷三磷酸及其相关产品的需求是一个数十亿美元的行业。高价值和相对较小的产量非常适合NN-Labs，这是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进入这个市场。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--费耶特维尔阿肯色大学的大力支持下，在这个传统上属于农村的州发展以知识为基础的产业。与阿肯色大学通过分包合同进行的拟议合作将为阿肯色大学的学生提供在高科技行业接触的独特机会。

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.