Biocatalysis and Bioprocessing Group Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus, Denmark.
Enzyme Engineering Group Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus, Denmark.
Chembiochem. 2022 Dec 5;23(23):e202200482. doi: 10.1002/cbic.202200482. Epub 2022 Nov 10.
Since its discovery in 2017, the fatty acid decarboxylase (FAP) photoenzyme has been the focus of extensive research, given its ability to convert fatty acids into alka(e)nes using merely visible blue light. Unfortunately, there are still some drawbacks that limit the applicability of this biocatalyst, such as poor solubility of the substrates in aqueous media, poor photostability, and the impossibility of reusing the catalyst for several cycles. In this work, we demonstrate the use of FAP in non-conventional media as a free enzyme and an immobilized preparation. Namely, its applicability in deep eutectic solvents (DESs) and a proof-of-concept immobilization using a commercial His-tag selective carrier, a thorough study of reaction and immobilization conditions in each case, as well as reusability studies are shown. We observed an almost complete selectivity of the enzyme towards C18 decarboxylation over C16 when used in a DES, with a product analytical yield up to 81 % when using whole cells. Furthermore, when applying the immobilized enzyme in DES, we obtained yields >10-fold higher than the ones obtained in aqueous media.
自 2017 年发现以来，脂肪酸脱羧酶（FAP）光酶一直是广泛研究的焦点，因为它仅使用可见光蓝色光就能将脂肪酸转化为链烷烃。然而，该生物催化剂仍存在一些限制其适用性的缺点，例如底物在水介质中的溶解度差、光稳定性差以及无法重复使用催化剂进行多个循环。在这项工作中，我们展示了 FAP 在非传统介质中作为游离酶和固定化制剂的用途。也就是说，它在深共熔溶剂（DES）中的适用性以及使用商业 His 标签选择性载体进行的概念验证固定化，对每种情况下的反应和固定化条件进行了全面研究，并进行了可重复使用性研究。我们观察到，当在 DES 中使用时，该酶对 C18 脱羧的选择性几乎完全超过 C16，当使用全细胞时，产物分析收率高达 81%。此外，当在 DES 中应用固定化酶时，我们获得的产率比在水介质中获得的产率高 10 倍以上。
