Ko_2021_Microb.Cell.Fact_20_232

Reference

Title : A novel protein fusion partner, carbohydrate-binding module family 66, to enhance heterologous protein expression in Escherichia coli - Ko_2021_Microb.Cell.Fact_20_232
Author(s) : Ko H , Kang M , Kim MJ , Yi J , Kang J , Bae JH , Sohn JH , Sung BH
Ref : Microb Cell Fact , 20 :232 , 2021
Abstract :

BACKGROUND: Proteins with novel functions or advanced activities developed by various protein engineering techniques must have sufficient solubility to retain their bioactivity. However, inactive protein aggregates are frequently produced during heterologous protein expression in Escherichia coli. To prevent the formation of inclusion bodies, fusion tag technology has been commonly employed, owing to its good performance in soluble expression of target proteins, ease of application, and purification feasibility. Thus, researchers have continuously developed novel fusion tags to expand the expression capacity of high-value proteins in E. coli. RESULTS: A novel fusion tag comprising carbohydrate-binding module 66 (CBM66) was developed for the soluble expression of heterologous proteins in E. coli. The target protein solubilization capacity of the CBM66 tag was verified using seven proteins that are poorly expressed or form inclusion bodies in E. coli: four human-derived signaling polypeptides and three microbial enzymes. Compared to native proteins, CBM66-fused proteins exhibited improved solubility and high production titer. The protein-solubilizing effect of the CBM66 tag was compared with that of two commercial tags, maltose-binding protein and glutathione-S-transferase, using poly(ethylene terephthalate) hydrolase (PETase) as a model protein; CBM66 fusion resulted in a 3.7-fold higher expression amount of soluble PETase (approximately 370smg/L) compared to fusion with the other commercial tags. The intact PETase was purified from the fusion protein upon serial treatment with enterokinase and affinity chromatography using levan-agarose resin. The bioactivity of the three proteins assessed was maintained even when the CBM66 tag was fused. CONCLUSIONS: The use of the CBM66 tag to improve soluble protein expression facilitates the easy and economic production of high-value proteins in E. coli.

PubMedSearch : Ko_2021_Microb.Cell.Fact_20_232
PubMedID: 34963459

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Citations formats

Ko H, Kang M, Kim MJ, Yi J, Kang J, Bae JH, Sohn JH, Sung BH (2021)
A novel protein fusion partner, carbohydrate-binding module family 66, to enhance heterologous protein expression in Escherichia coli
Microb Cell Fact 20 :232

Ko H, Kang M, Kim MJ, Yi J, Kang J, Bae JH, Sohn JH, Sung BH (2021)
Microb Cell Fact 20 :232