Manuel Brea A, Das Soumen, Sanford Aimee, Jenkins Matthew C, Finn M G, Heemstra Jennifer M
Department of Chemistry, Emory University, Atlanta, Georgia 30322, United States.
School of Chemistry and Biochemistry, Georgia Institute of Technology, 901 Atlantic Drive, Atlanta, Georgia 30306, United States.
Biomacromolecules. 2023 Apr 10;24(4):1934-1941. doi: 10.1021/acs.biomac.3c00144. Epub 2023 Mar 29.
Enzyme activity requires sequential binding and chemical transformation of substrates. While directed evolution and random mutagenesis are common methods for improving catalytic activity, these methods do not allow for independent control of and . To achieve such control, we envisioned that the colocalization of aptamers and enzymes that act on the same molecule could increase catalytic efficiency through preconcentration of substrate. We explored this concept with cocaine esterase and anticocaine aptamers having varying values, both encapsulated in MS2 virus-like particles. Rate enhancements were observed with magnitudes dependent on both aptamer:enzyme stoichiometry and aptamer , peaking when aptamer and enzyme were roughly equivalent. This beneficial effect was lost when either aptamer binding was too tight or the aptamers were not constrained to be close to the catalyst. This work demonstrates a modular way to enhance catalysis by independently controlling substrate capture and release to the processing enzyme.
酶活性需要底物的顺序结合和化学转化。虽然定向进化和随机诱变是提高催化活性的常用方法，但这些方法无法独立控制[具体物质1]和[具体物质2]。为实现这种控制，我们设想作用于同一分子的适体和酶的共定位可以通过底物的预浓缩提高催化效率。我们用包裹在MS2病毒样颗粒中的具有不同[相关参数]值的可卡因酯酶和抗可卡因适体探索了这一概念。观察到速率增强，其幅度取决于适体与酶的化学计量比和适体[相关参数]，当适体[相关参数]和酶[相关参数]大致相等时达到峰值。当适体结合过紧或适体未被限制在靠近催化剂的位置时，这种有益效果就会丧失。这项工作展示了一种通过独立控制底物向加工酶的捕获和释放来增强催化作用的模块化方法。
