Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA.
Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
Angew Chem Int Ed Engl. 2016 Jul 18;55(30):8585-9. doi: 10.1002/anie.201602353. Epub 2016 May 30.
Enzyme-mediated protein modification often requires large amounts of biocatalyst, adding significant costs to the process and limiting industrial applications. Herein, we demonstrate a scalable and straightforward strategy for the efficient capture and recycling of enzymes using a small-molecule affinity tag. A proline variant of an evolved sortase A (SrtA 7M) was N-terminally labeled with lithocholic acid (LA)-an inexpensive bile acid that exhibits strong binding to β-cyclodextrin (βCD). Capture and recycling of the LA-Pro-SrtA 7M conjugate was achieved using βCD-modified sepharose resin. The LA-Pro-SrtA 7M conjugate retained full enzymatic activity, even after multiple rounds of recycling.
酶介导的蛋白质修饰通常需要大量的生物催化剂，这会增加工艺成本，并限制工业应用。在此，我们展示了一种使用小分子亲和标签高效捕获和回收酶的可扩展且简单的策略。一种经过进化的 sortase A（SrtA 7M）的脯氨酸变体在 N 端用石胆酸（LA）标记 - 一种廉价的胆汁酸，它与 β-环糊精（βCD）具有很强的结合能力。使用 βCD 修饰的琼脂糖树脂实现 LA-Pro-SrtA 7M 缀合物的捕获和回收。LA-Pro-SrtA 7M 缀合物即使经过多次回收循环，仍保持完全的酶活性。
