Graz University of Technology, Institute of Molecular Biotechnology, Petersgasse 14, 8010, Graz, Austria.
Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50, 8010, Graz, Austria.
Sci Rep. 2019 Feb 25;9(1):2697. doi: 10.1038/s41598-019-39484-9.
The enzyme 4-oxalocrotonate tautomerase shows remarkable catalytic versatility due to the secondary amine of its N-terminal proline moiety. In this work, we incorporated a range of proline analogues into the enzyme and examined the effects on structure and activity. While the structure of the enzyme remained unperturbed, its promiscuous Michael-type activity was severely affected. This finding demonstrates how atomic changes in a biocatalytic system can abolish its activity. Our work provides a toolbox for successful generation of enzyme variants with non-canonical catalytic proline analogues.
由于其 N 端脯氨酸部分的仲胺，4-氧代戊烯二酸反式酶表现出显著的催化多功能性。在这项工作中，我们将一系列脯氨酸类似物掺入到酶中，并研究了它们对结构和活性的影响。虽然酶的结构保持不变，但它的混杂迈克尔型活性受到严重影响。这一发现表明生物催化系统中的原子变化如何能使其活性丧失。我们的工作为成功生成具有非典型催化脯氨酸类似物的酶变体提供了一个工具箱。
