Department of Chemistry and Biochemistry, ‡Center for Bio-Inspired Nanomaterials, and §Deparment of Mathematical Sciences, Montana State University , Bozeman, Montana 59717, United States.
ACS Chem Biol. 2014 Feb 21;9(2):359-65. doi: 10.1021/cb4006529. Epub 2013 Dec 6.
Developing methods for investigating coupled enzyme systems under conditions that mimic the cellular environment remains a significant challenge. Here we describe a biomimetic approach for constructing densely packed and confined multienzyme systems through the co-encapsulation of 2 and 3 enzymes within a virus-like particle (VLP) that perform a coupled cascade of reactions, creating a synthetic metabolon. Enzymes are efficiently encapsulated in vivo with known stoichiometries, and the kinetic parameters of the individual and coupled activities are characterized. From the results we develop and validate a mathematical model for predicting the expected kinetics for coupled reactions under co-localized conditions.
开发在模拟细胞环境的条件下研究偶联酶系统的方法仍然是一个重大挑战。在这里，我们描述了一种仿生方法，通过将两种和三种酶共同包封在病毒样颗粒（VLP）中，构建紧密堆积和受限的多酶系统，从而进行偶联级联反应，创建合成代谢物。酶以已知的化学计量比在体内有效地被包封，并且个体和偶联活性的动力学参数得到了表征。根据这些结果，我们开发并验证了一个数学模型，用于预测在共定位条件下偶联反应的预期动力学。
