Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, National Engineering Laboratory for Industrial Enzymes, Tianjin, China.
Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
J Sci Food Agric. 2024 Jan 15;104(1):266-272. doi: 10.1002/jsfa.12907. Epub 2023 Aug 22.
Compound 2-O-α-d-glucosylglycerol (2αGG) naturally serves as a compatible osmolyte in acclimation to environmental stresses, such as high osmolarity, dryness, and extreme temperature. It presents several bioactivities and has been used in the food, agriculture, and cosmetics areas.
In the present study, we attempted to synthesize the 2αGG from low-cost maltodextrin and glycerol by constructing an in vitro multi-enzyme system. The system contained two core enzymes, namely glucan phosphorylases (GPs) and glucosylglycerol phosphorylases (GGPs), and two auxiliary enzymes, namely isoamylase and 4-α-glucanotransferase. Several new GGPs from different organisms were characterized with the function of converting α-G1P and glycerol to sole stereo-configuration product 2αGG. Then, polypeptide SpyTag-SpyCatcher was employed to construct a self-assembled multienzyme complex, and different combinations between enzymes and peptides were constructed and tested. The best self-assembled multienzyme complex exhibited three-fold higher productivity compared to that of free enzyme. This reaction system also produced 240 mm (61 g L ) 2αGG under high substrate concentration, with a conversion yield of 86%.
The present study provides an efficient approach for producing 2αGG. It also demonstrates that the SpyTag-SpyCatcher system could be applied to construct other multienzyme complexes for increased productivity and product titer. © 2023 Society of Chemical Industry.
2-O-α-d-葡萄糖基甘油（2αGG）作为一种天然的相容性渗透剂，在适应环境压力（如高渗透压、干燥和极端温度）方面具有多种生物活性，已在食品、农业和化妆品领域得到应用。
本研究试图利用廉价的麦芽糊精和甘油构建体外多酶体系来合成 2αGG。该体系包含两种核心酶，即葡聚糖磷酸化酶（GPs）和甘油葡糖苷磷酸化酶（GGPs），以及两种辅助酶，即异淀粉酶和 4-α-葡聚糖转移酶。几种来自不同生物体的新型 GGPs 被表征为具有将 α-G1P 和甘油转化为单一立体构型产物 2αGG 的功能。然后，采用多肽 SpyTag-SpyCatcher 构建自组装多酶复合物，并构建和测试了不同酶与肽之间的组合。最佳的自组装多酶复合物的产物比游离酶高 3 倍。该反应体系在高底物浓度下可产生 240 mm（61 g L-1）的 2αGG，转化率为 86%。
本研究为 2αGG 的生产提供了一种有效的方法。还表明 SpyTag-SpyCatcher 系统可用于构建其他多酶复合物，以提高生产力和产物浓度。© 2023 化学工业协会。
