Efficient and Scalable Production Efficient and Scalable Production of Biomolecular Materials Using Enzymatic Oxidative Coupling Strategies

生物分子和合成材料之间的界面是一个重要的研究前沿，在医学诊断、疾病治疗、病原体检测和监测以及工业催化方面具有潜在的应用前景。这些应用的关键是获得有效的化学技术，将合成成分附着到复杂的蛋白质和核酸表面的特定位置。尽管已经有一些经过时间考验的战略用于这一目的，但它们通常难以控制，大规模使用的成本也很高。此外，这些策略中的大多数会产生复杂的产品混合物，导致材料定义不清。在这个由化学系大分子、超分子和纳米化学计划资助的项目中，加州大学伯克利分校的Matthew B.Francis将开发一种新的基于酶的反应，用于在合成材料中结合蛋白质和核酸。与其他方法相比，这项技术可以在单一位置修饰复杂的蛋白质，并依赖于任何所需规模的廉价构建块。该计划产生了一种强大的技术，可以通过定义明确的连接将任何蛋白质与几乎任何感兴趣的材料结合起来。该计划支持的外展活动激发了当地学校对K-5学生的科学兴趣。正在开发以“厨房里的化学和生物”为基础的课堂演示。演讲将包括使用棉花糖、巧克力、蓝莓和其他孩子喜欢的食物进行令人难忘的、引人入胜的化学概念演示。本程序中的化学基于一系列氧化偶联反应，这些反应生成邻苯二酚和邻亚氨基酚中间体。这些中间体可以与各种材料的苯胺、N-末端脯氨酸和游离半胱氨酸侧链以非常高的速率反应，生成高度稳定的连接物。这类反应已被发现与附着在聚合物、生色团和纳米颗粒上的各种生物官能团一起工作。以前，这个反应是通过添加氧化剂，如铁氰化物或高碘酸盐来引发的。我们探索了一种酶催化的方法，该方法以大气中的氧气为氧化剂，以水为唯一副产品，具有类似的效率。该计划的具体目标包括(1)展示该技术在一系列有趣的蛋白质底物上的应用，(2)优化反应的酶成分，以提供更高的催化效率和储存稳定性，以及(3)展示新化学在纸和棉花等纤维素底物的改性中的应用，以生产用于水净化的低成本、可生物降解的材料。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The interface between biological molecules and synthetic materials represents a major research frontier that has potential applications in medical diagnostics, disease treatment, pathogen detection and monitoring, and industrial catalysis. Key to these applications is the availability of efficient chemical technologies for the attachment of synthetic components to specific locations on complex protein and nucleic acid surfaces. Although a number of time-tested strategies already exist for this purpose, they are generally difficult to control and costly to use on large scale. Additionally, most of these strategies generate complex mixtures of products leading to poorly-defined materials. In this project funded by the Macromolecular, Supramolecular and Nanochemistry Program of the Chemistry Division, Matthew B. Francis of the University of California, Berkeley, will develop a new enzyme-based reaction for the incorporation of proteins and nucleic acids in synthetic materials. Relative to other methods, this technique can modify complex proteins in single locations and relies on inexpensive building blocks that are available on any desired scale. The program yields a powerful technology that can be used to combine any protein with virtually any material of interest through well-defined linkages. Outreach activities supported by this program spark science interest in K-5 students in local schools. Classroom presentations based on "Chemistry and Biology in the Kitchen" are being developed. The presentations will include memorable and engaging demonstrations of chemical concepts using marshmallows, chocolate, blue berries, and other foods kids love. The chemistry in this program is based on a series of oxidative coupling reactions that generate ortho-quinone and ortho-iminoquinone intermediates. These intermediates can react with anilines, N-terminal prolines, and free cysteine side chains of various materials with very high rates to yield highly stable conjugates. This class of reactions has been found to work with a full range of biological functional groups attached to polymers, chromophores, and nanoparticles. Previously, this reaction was initiated by the addition of oxidants, such as ferricyanide or periodate. We explore an enzyme-catalyzed method that has similar efficiency using atmospheric oxygen as the oxidant and producing water as the sole byproduct. Specific goals of this program include (1) demonstrating the utility of the technique on a series of interesting protein substrates, (2) optimizing the enzyme component of the reaction to afford greater catalytic efficiency and storage stability, and (3) showcasing the application of the new chemistry to the modification of cellulosic substrates, such as paper and cotton to produce low-cost, biodegradable materials for water purification.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.