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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
PubMed

Insights

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.
Keywords:
AMPAntibiotic resistanceArenicinRecombinant expressionSUMO

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