高度稳定且可重复使用的固定化甲酸脱氢酶：用于NADH原位再生的有前景的生物催化剂。

Highly stable and reusable immobilized formate dehydrogenases: Promising biocatalysts for in situ regeneration of NADH.

作者信息

Binay Barış, Alagöz Dilek, Yildirim Deniz, Çelik Ayhan, Tükel S Seyhan

机构信息

Istanbul AREL University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, Tepekent, Büyükcekmece, Istanbul, Turkey.

University of Cukurova, Vocational School of Imamoglu, Adana, Turkey.

出版信息

Beilstein J Org Chem. 2016 Feb 12;12:271-7. doi: 10.3762/bjoc.12.29. eCollection 2016.

This study aimed to prepare robust immobilized formate dehydrogenase (FDH) preparations which can be used as effective biocatalysts along with functional oxidoreductases, in which in situ regeneration of NADH is required. For this purpose, Candida methylica FDH was covalently immobilized onto Immobead 150 support (FDHI150), Immobead 150 support modified with ethylenediamine and then activated with glutaraldehyde (FDHIGLU), and Immobead 150 support functionalized with aldehyde groups (FDHIALD). The highest immobilization yield and activity yield were obtained as 90% and 132%, respectively when Immobead 150 functionalized with aldehyde groups was used as support. The half-life times (t 1/2) of free FDH, FDHI150, FDHIGLU and FDHIALD were calculated as 10.6, 28.9, 22.4 and 38.5 h, respectively at 35 °C. FDHI150, FDHIGLU and FDHIALD retained 69, 38 and 51% of their initial activities, respectively after 10 reuses. The results show that the FDHI150, FDHIGLU and FDHIALD offer feasible potentials for in situ regeneration of NADH.

本研究旨在制备稳定的固定化甲酸脱氢酶（FDH）制剂，其可与功能性氧化还原酶一起用作有效的生物催化剂，其中需要NADH的原位再生。为此，将甲基假丝酵母FDH共价固定在Immobead 150载体上（FDHI150），将Immobead 150载体用乙二胺改性，然后用戊二醛活化（FDHIGLU），以及将Immobead 150载体用醛基官能化（FDHIALD）。当使用用醛基官能化的Immobead 150作为载体时，固定化产率和活性产率分别最高，为90%和132%。在35℃下，游离FDH、FDHI150、FDHIGLU和FDHIALD的半衰期（t1/2）分别计算为10.6、28.9、22.4和38.5小时。FDHI150、FDHIGLU和FDHIALD在重复使用10次后，分别保留其初始活性的69%、38%和51%。结果表明，FDHI150、FDHIGLU和FDHIALD为NADH的原位再生提供了可行的潜力。