Soluble expression of hMYDGF was improved by strain engineering and optimizations of fermentation strategies in Escherichia coli.

Autor: Su SY; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., Zheng YS; 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., Mao H; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., Zhao LB; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., Zhu MY; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., Yang YF; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., Li LT; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., Wang ZR; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China., He C; Shanghai Institute of Biological Products Co., Ltd., Shanghai, China. Electronic address: hcgasurada@live.com.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2024 Dec; Vol. 224, pp. 106565. Date of Electronic Publication: 2024 Aug 05.
DOI: 10.1016/j.pep.2024.106565
Abstrakt: 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.
(Copyright © 2024 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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