Department of Life Science and Biochemical Engineering, Sun Moon University, 70 Sunmoon-ro 221, Tangjeong-myeon, Asan-si 31460, Chungnam, Korea.
Department of Pharmaceutical Engineering and Biotechnology, Sun Moon University, 70 Sunmoon-ro 221, Tangjeong-myeon, Asan-si 31460, Chungnam, Korea.
Molecules. 2022 May 25;27(11):3405. doi: 10.3390/molecules27113405.
is traditionally known as a food-grade microorganism due to its high ability to produce amino acids and its endotoxin-free recombinant protein expression factory. In recent years, studies to improve the activities of useful therapeutics and pharmaceutical compounds have led to the engineering of the therapeutically advantageous cell factory system. One of the well-studied ways to improve the activities of useful compounds is glucosylation with glycosyltransferases. In this study, we successfully and efficiently glycosylated therapeutic butyl-4-aminobenzoate and other -linked compounds in using a promiscuous YdhE, which is a glycosyltransferase from . For efficient glucosylation, components, such as promoter, codons sequence, expression temperatures, and substrate and glucose concentrations were optimized. With glucose as the sole carbon source, we achieved a conversion rate of almost 96% of the glycosylated products in the culture medium. The glycosylated product of high concentration was successfully purified by a simple purification method, and subjected to further analysis. This is a report of the in vivo cultivation and glucosylation of -linked compounds in .
由于其生产氨基酸的能力很高，且能无内毒素地表达重组蛋白，所以传统上它被认为是一种食品级微生物。近年来，为了提高有用治疗药物和药物化合物的活性，人们对治疗有益的细胞工厂系统进行了工程设计。提高有用化合物活性的一种研究得很好的方法是用糖基转移酶进行糖基化。在这项研究中，我们使用一种从 中获得的杂合 YdhE 成功且有效地对治疗性丁酸-4-氨基苯甲酸和其他 - 连接的化合物进行了糖基化。为了实现高效糖基化，我们对启动子、密码子序列、表达温度以及底物和葡萄糖浓度等成分进行了优化。在仅以葡萄糖为碳源的情况下，我们在培养基中实现了近 96%的糖基化产物转化率。通过一种简单的纯化方法，成功地对高浓度的糖基化产物进行了纯化，并进一步进行了分析。这是在 中进行体内培养和 - 连接化合物糖基化的报告。
