College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, Jiangsu 211816, People's Republic of China.
School of Chemistry and Molecular Engineering, Nanjing Tech University, Nanjing, Jiangsu 210009, People's Republic of China.
J Agric Food Chem. 2022 Apr 20;70(15):4690-4700. doi: 10.1021/acs.jafc.2c01118. Epub 2022 Apr 11.
Trehalose is an important rare sugar that protects biomolecules against environmental stress. We herein introduce a dual enzyme cascade strategy that regulates the proportion of cargos and scaffolds, to maximize the benefits of enzyme immobilization. Based upon the self-assembling properties of the shell protein (EutM) from the ethanolamine utilization (Eut) bacterial microcompartment, we implemented the catalytic synthesis of trehalose from soluble starch with the coimmobilization of α-amylase and trehalose synthase. This strategy improved enzymatic cascade activity and operational stability. The cascade system enabled the efficient production of trehalose with a yield of ∼3.44 g/(L U), 1.5 times that of the free system. Moreover, its activity was maintained over 12 h, while the free system was almost completely inactivated after 4 h, demonstrating significantly enhanced thermostability. In conclusion, an attractive self-assembly coimmobilization platform was developed, which provides an effective biological process for the enzymatic synthesis of trehalose .
海藻糖是一种重要的稀有糖，可保护生物分子免受环境压力的影响。我们在此介绍了一种双酶级联策略，该策略可调节货物和支架的比例，以最大程度地提高酶固定化的益处。基于乙醇胺利用（Eut）细菌微隔间中壳蛋白（EutM）的自组装特性，我们实现了用α-淀粉酶和海藻糖合酶共固定化从可溶性淀粉催化合成海藻糖。该策略提高了酶级联反应的活性和操作稳定性。该级联系统可有效地生产海藻糖，产率约为 3.44 g/（L U），是游离系统的 1.5 倍。此外，其活性可维持 12 小时以上，而游离系统在 4 小时后几乎完全失活，表明其热稳定性显著提高。总之，开发了一种有吸引力的自组装共固定化平台，为海藻糖的酶促合成提供了一种有效的生物过程。
