Novel DNA-Based Biocatalytic Nanomaterials

ID：MPS/DMR/BMAT(7623)1005609 PI：Kumar，Challa ORG：康涅狄格州大学题目：新型脱氧核糖核酸生物催化纳米材料特长：这项建议开发了一种将酶连接到固体底物上的策略，用于各种应用，在这些应用中，可以利用酶的高化学、区域、立体和手性选择性。由于结合在固体上的酶通常比相应的游离酶活性低，所以PI将首先研究酶与层状固体如α-磷酸-锆磷酸盐(α-ZrRPs)结合的机制。系统地改变α-ZrRP的表面函数(R)来控制这些相互作用，PI将量化伴随结合事件的特定过程的焓和熵贡献的性质和大小，以阐明酶-固体相互作用的分子特征。利用这些洞察力，他将通过DNA介导的自组装，在无试剂的条件下制备合理设计的酶-DNA-ZrRP偶联物。该方法包括：(1)通过长绳将单链DNA共价连接到特定的ZrRP纳米颗粒上；(2)将互补的单链DNA连接到表面的COOH基团上；(3)将酶-DNA与DNA-ZrRP杂交，制备酶-DNA-纳米材料。系链将由低聚乙二醇基(OEG)间隔基组成，因为已知这些间隔基与蛋白质或DNA几乎没有或几乎没有相互作用。DNA杂交是众所周知的，它应该在无试剂的条件下产生酶-DNA-纳米颗粒。DNA在固体和蛋白质之间提供了强大的、可逆的、间接的联系。DNA标记的酶和纳米粒子将为各种实际应用提供大量机会来构建更高阶的组件。BROADER的影响：在研究生水平，PI将继续开发两门新课程(生物化学I和II，化学360/361，每门3学分)，旨在培训学生将化学原理应用于重要的生物系统。生物电子传递、生物材料、生物量热学和DNA/蛋白质结构是PI为这些课程贡献的一些主题。PI正在开发技术交流、写作和道德方面的第三门研究生课程。在本科阶段，PI将提供生物材料、蛋白质无机材料和DNA无机材料的研究培训。在过去的9年里，作为化学系实习计划的一部分，近30名本科生在PI的实验室获得了这样的研究经验。参与的本科生经常被列为结果出版物的共同作者。在高中阶段，PI参加由康涅狄格大学赞助的全大学范围内的优秀高中生导师联系计划。他还接待中学生参加为期一天的动手实验室体验，供来自康涅狄格州各地的学生使用。

ID: MPS/DMR/BMAT(7623) 1005609 PI: Kumar, Challa ORG: University of ConnecticutTitle: Novel DNA-Based Biocatalytic NanomaterialsINTELLECTUAL 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.