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High-yield and cost-effective biosynthesis process for producing antimicrobial peptide AA139
Ying Zhang1, Yapeng Wang1, Jianguang Lu2
1School of Pharmacy, Fudan University, Shanghai, 201203, People's Republic of China; Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, Shanghai, 201203, People's Republic of China.
Protein Expression and Purification
|March 29, 2024
Summary
This study developed a novel bioprocess for producing AA139, a potent antimicrobial peptide (AMP), overcoming previous yield limitations. The optimized method achieved significantly higher yields of AA139 for potential clinical applications against resistant bacteria.
Area of Science:
- Biotechnology
- Microbiology
- Biochemistry
Background:
- AA139, a potent antimicrobial peptide (AMP) variant, shows activity against multidrug-resistant (MDR) and extensively drug-resistant (XDR) Gram-negative bacteria.
- Previous production methods for AA139 yielded less than 5 mg/L and were hindered by complex chemical synthesis, limiting research and clinical use.
Purpose of the Study:
- To establish an efficient bioprocess for producing AA139.
- To significantly increase the yield and purity of recombinant AA139 (rAA139) for further investigation and potential clinical applications.
Main Methods:
- AA139 was expressed intracellularly in Escherichia coli (E. coli) BL21 (DE3) using SUMO fusion technology.
- A simplified downstream process was developed based on physicochemical characteristics.
- High cell density fermentation (HCDF) was optimized to enhance protein expression by 54%.
Main Results:
- A yield of 56 mg of rAA139 per liter of culture was achieved with 98% purity, the highest reported to date.
- Characterization confirmed the molecular mass, disulfide bonds, and antimicrobial activity of the produced rAA139.
- The developed bioprocess overcomes previous limitations in AA139 production.
Conclusions:
- The study presents a highly efficient bioprocess for producing recombinant AA139.
- This optimized production method significantly increases yield and purity, facilitating future research and clinical development of AA139 as an antimicrobial agent.
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