Polymer-Based Protein Engineering Enables Molecular Dissolution of Chymotrypsin in Acetonitrile
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- 作者：Chad S. Cummings ; Hironobu Murata ; Krzysztof Matyjaszewski ; Alan J. Russell
- 刊名：ACS Macro Letters
- 出版年：2016
- 出版时间：April 19, 2016
- 年：2016
- 卷：5
- 期：4
- 页码：493-497
- 全文大小：264K
- 年卷期：0
- ISSN：2161-1653
文摘
While most effective in aqueous environments, enzymes are also able to catalyze reactions in essentially anhydrous organic media. Enzyme activity in organic solvents is limited as a result of inefficient substrate binding, lack of solubility, and inactivation by hydrophilic anhydrous solvents. With these facts in mind, atom transfer radical polymerization was used to synthesize chymotrypsin-poly(2-(dimethylamino)ethyl methacrylate) (CT-pDMAEMA) conjugates designed to be soluble and active in acetonitrile. CT-pDMAEMA solubility in organic solvents and the rate of CT-pDMAEMA-catalyzed transesterification in acetonitrile were examined at a range of water (0–15 M) and propanol (0.01–5 M) concentrations. The conjugates were soluble at the molecular scale in several organic solvents, exhibited good substrate binding with N-acetyl l-phenylalanine thiophenylester (KM as low as 17 mM), and had an activity (peak activity 330 μM/min/mg enzyme) many orders of magnitude higher than that of the insoluble native enzyme.