Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, College of Life Sciences, Jilin University, Changchun 130012, People's Republic of China.
Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, College of Life Sciences, Jilin University, Changchun 130012, People's Republic of China.
Enzyme Microb Technol. 2018 Feb;109:66-73. doi: 10.1016/j.enzmictec.2017.09.013. Epub 2017 Sep 29.
Herein we established co-immobilized methods for enoate reductases (ERs) and glucose dehydrogenase (GDH), forming a cofactor regeneration system. In cross-linked enzyme aggregates (CLEAs), ammonium sulfate and oxidized dextran were selected as a precipitant and a cross-linker, respectively. In biomimetic immobilization (BI), ER-GDH-silica particles (ER-GDH-SPs) were rapidly formed through a one-step approach by using a silicic acid precursor. Under the optimal conditions, the ER activity recovery in ER-GDH-CLEAs and ER-GDH-SPs were 44.9±1.8% and 44.5±2.1%, and the immobilization efficiency was 93.5±1.2% and 92.4±1.2%, respectively. ER-GDH-CLEAs and ER-GDH-SPs exhibit excellent thermal and pH stability, and superior reusability. The activity of ER-GDH-SPs toward the substrate is also better than that of free ER and GDH in reduction of 4-(4-Methoxyphenyl)-3-buten-2-one. This study introduces simple and inexpensive co-immobilization strategies to construct novel and efficient ER-GDH-CLEAs and ER-GDH-SPs with high activity and stability.
在此，我们建立了烯醇还原酶（ER）和葡萄糖脱氢酶（GDH）的共固定化方法，形成了辅酶再生系统。在交联酶聚集体（CLEAs）中，硫酸铵和氧化葡聚糖分别被选为沉淀剂和交联剂。在仿生固定化（BI）中，通过使用硅酸前体，通过一步法快速形成 ER-GDH-二氧化硅颗粒（ER-GDH-SPs）。在最佳条件下，ER-GDH-CLEAs 和 ER-GDH-SPs 中的 ER 活性回收率分别为 44.9±1.8%和 44.5±2.1%，固定化效率分别为 93.5±1.2%和 92.4±1.2%。ER-GDH-CLEAs 和 ER-GDH-SPs 具有出色的热稳定性和 pH 稳定性，以及优异的可重复使用性。与游离 ER 和 GDH 相比，ER-GDH-SPs 对 4-（4-甲氧基苯基）-3-丁烯-2-酮的底物的活性也更好。本研究介绍了简单且廉价的共固定化策略，以构建具有高活性和稳定性的新型高效 ER-GDH-CLEAs 和 ER-GDH-SPs。
