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The Multifaceted Benefits of Protein Co-expression in Escherichia coli
Published on: February 5, 2015
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Engineering versatile protein expression systems mediated by inteins in Escherichia coli
Keith W Y Kwong1, Alan K L Ng1, W K R Wong2
1Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
Applied Microbiology and Biotechnology
|September 19, 2015
Summary
Inteins enable efficient co-expression of multiple proteins in Escherichia coli. This study shows VMA and GyrA inteins facilitate the production of human epidermal growth factor (EGF) and endoglucanase (Eng) in E. coli, yielding authentic products in both cytoplasm and media.
Area of Science:
- Biotechnology
- Molecular Biology
- Protein Engineering
Background:
- Intein-mediated protein ligation offers a powerful tool for protein engineering.
- Previous work demonstrated the utility of Saccharomyces cerevisiae vascular membrane ATPase (VMA) intein for co-expressing human epidermal growth factor (EGF) and human basic fibroblast growth factor (bFGF) in Escherichia coli.
- The VMA intein system facilitated secretion and cytoplasmic localization of both EGF and bFGF.
Purpose of the Study:
- To investigate the efficacy of two structurally distinct inteins, VMA and Mycobacterium xenopi GyraseA (GyrA), in mediating the co-expression of disparate proteins.
- To assess the ability of these inteins to facilitate the production of authentic, soluble, and auto-cleavable protein products in Escherichia coli.
- To further validate the potential of intein-based Escherichia coli systems for cost-effective and versatile protein production.
Main Methods:
- Engineered Escherichia coli strains expressing VMA and GyrA inteins.
- Fusion of human epidermal growth factor (EGF) and endoglucanase (Eng) genes to the 5' and 3' ends of the intein genes, respectively.
- Analysis of protein expression, solubility, auto-cleavage, and localization in both the cytoplasm and culture medium.
Main Results:
- Both VMA and GyrA inteins successfully mediated the co-expression of EGF and Eng in Escherichia coli.
- The expressed proteins, initially in precursor forms, were soluble and auto-cleavable, yielding authentic EGF and Eng.
- The desired products were recovered from both the bacterial cytoplasm and the culture medium.
Conclusions:
- Inteins, including VMA and GyrA, are effective in mediating the co-expression of diverse proteins in Escherichia coli.
- The intein/E. coli system facilitates the production of soluble, authentic proteins that are accessible in both cellular and extracellular fractions.
- This approach holds promise for developing versatile and economical host systems for recombinant protein production.
