Shanghai Institute of Biological Products Co., Ltd., Shanghai, China.
Shanghai Institute of Biological Products Co., Ltd., Shanghai, China; Research Center for Translational Medicine at East Hospital, School of Life Sciences and Technology, Tongji University, Shanghai, China.
Protein Expr Purif. 2024 Dec;224:106565. doi: 10.1016/j.pep.2024.106565. Epub 2024 Aug 5.
Myeloid-derived growth factor (MYDGF) is a cytokine that exhibits a variety of biological functions. This study focused on utilizing BL21(DE3) strain engineering and fermentation strategies to achieve high-level expression of soluble human MYDGF (hMYDGF) in Escherichia coli. Initially, the E. coli expressing strain BL21(DE3) was engineered by deleting the IpxM gene and inserting the GROEL/S and Trigger factor genes. The engineered E. coli strain BL21(TG)/pT-MYDGF accumulated 3557.3 ± 185.6 μg/g and 45.7 ± 6.7 mg/L of soluble hMYDGF in shake flask fermentation, representing a 15.6-fold increase compared to the control strain BL21(DE3)/pT-MYDGF. Furthermore, the yield of hMYDGF was significantly enhanced by optimizing the fermentation conditions. Under optimized conditions, the 5L bioreactor yielded up to 2665.8 ± 164.3 μg/g and 407.6 ± 42.9 mg/L of soluble hMYDGF. The results indicate that the implementation of these optimization strategies could enhance the ratio and yield of soluble proteins expressed by E.coli, thereby meeting the demands of industrial production. This study employed sophisticated strategies to lay a solid foundation for the industrial application of hMYDGF.
髓系细胞衍生生长因子（MYDGF）是一种细胞因子，具有多种生物学功能。本研究旨在利用 BL21（DE3）菌株工程和发酵策略，在大肠杆菌中实现可溶性人 MYDGF（hMYDGF）的高水平表达。首先，通过删除 IpxM 基因并插入 GROEL/S 和 Trigger factor 基因，对表达菌株 BL21（DE3）进行工程改造。工程菌 BL21（TG）/pT-MYDGF 在摇瓶发酵中积累了 3557.3±185.6μg/g 和 45.7±6.7mg/L 的可溶性 hMYDGF，与对照菌株 BL21（DE3）/pT-MYDGF 相比，增加了 15.6 倍。此外，通过优化发酵条件进一步提高了 hMYDGF 的产量。在优化条件下，5L 生物反应器可获得高达 2665.8±164.3μg/g 和 407.6±42.9mg/L 的可溶性 hMYDGF。结果表明，实施这些优化策略可以提高大肠杆菌表达的可溶性蛋白的比例和产量，从而满足工业生产的需求。本研究采用了复杂的策略，为 hMYDGF 的工业应用奠定了坚实的基础。
