Yang Liyan, Zhou Huimin, Chen Guiguang, Li Hongliang, Yang Dengfeng, Pan Lixia
State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Marine Natural Products and Combinatorial Biosynthesis Chemistry, Guangxi Academy of Sciences, Nanning 530007, China.
College of Life Science and Technology, Guangxi University, Nanning 530004, China.
Microorganisms. 2023 Mar 16;11(3):762. doi: 10.3390/microorganisms11030762.
Anthracyclines are an important class of natural antitumor drugs. They have a conservative aromatic tetracycline backbone that is substituted with different deoxyglucoses. The deoxyglucoses are crucial for the biological activity of many bacterial natural products after the proper modification from glycosyltransferases (GTs). The difficulty in obtaining highly purified active GTs has prevented biochemical studies on natural product GTs. In this paper, a new fusion plasmid pGro7', which introduces the chaperone genes , and was constructed. The glycosyltransferase DnmS from ATCC 27952 was co-expressed with the plasmid pGro7', and unprecedented high-efficiency and soluble expression of DnmS in the expression system was realized. Subsequently, the reverse glycosylation reaction characteristics of DnmS and DnmQ were verified. We found that DnmS and DnmQ had the highest enzyme activity when they participated in the reaction at the same time. These studies provide a strategy for the soluble expression of GTs in and confirm the reversibility of the catalytic reaction of GTs. This provides a powerful method for the production of active anthracyclines and to enhance the diversity of natural products.
蒽环类药物是一类重要的天然抗肿瘤药物。它们具有保守的芳香四环骨架，其上连接有不同的脱氧葡萄糖。在糖基转移酶（GTs）进行适当修饰后，脱氧葡萄糖对于许多细菌天然产物的生物活性至关重要。难以获得高度纯化的活性GTs阻碍了对天然产物GTs的生化研究。本文构建了一种新的融合质粒pGro7'，它引入了伴侣基因。来自ATCC 27952的糖基转移酶DnmS与质粒pGro7'共表达，实现了DnmS在表达系统中前所未有的高效且可溶性表达。随后，验证了DnmS和DnmQ的反向糖基化反应特性。我们发现DnmS和DnmQ同时参与反应时酶活性最高。这些研究为GTs在中的可溶性表达提供了一种策略，并证实了GTs催化反应的可逆性。这为活性蒽环类药物的生产以及增强天然产物的多样性提供了有力方法。
