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Optimization of plasmid electrotransformation into Bacillus subtilis using an antibacterial peptide

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Abstract

Bacillus subtilis can potentially serve as an efficient expression host for biotechnology due to its ability to secrete extracellular proteins and enzymes directly into the culture medium. One of the important challenges in the biotechnology industry is to optimize the transformation conditions of B. subtilis bacteria. This study aims to provide a new method to optimize the transformation conditions and improve the transformation efficiency of B. subtilis WB600. To increase the transformation efficiency in B. subtilis, two methods of adding CM11 antibacterial peptides to the bacterial medium along with electroporation and optimizing the variables including the growth medium composition, time to adding CM11 peptide, electroporation voltage, recovery medium, and cell recovery time are used. The results of this study showed that the addition of antimicrobial peptides (AMPs) with a concentration of 2 μg/ml increases the transformation efficiency by 4 times compared to the absence of AMP in the bacterial medium. Additionally, the findings from our study indicated that the most optimal rate of transformation for B. subtilis was observed at a voltage of 7.5 kV/cm, with a recovery period of 12 h. With the optimized method, the transformation efficiency came up to 1.69 × 104 CFU/µg DNA. This improvement in transformation efficiency will be attributed to the research of expression of exogenous genes in B. subtilis, gene library construction for transformation of wild-type B. subtilis strains.

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Data availability

The datasets generated and the data analysis during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors are grateful to Professor Reza Roosta Azad, the late professor of the Faculty of Chemical Engineering at Sharif University of Technology, for his efforts, assistance, and guidance in completing this project.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Department of Chemical and Petroleum Engineering, Sharif University of Technology, Tehran, Iran

    Mohamadsadegh Mohamadzadeh

  2. Rajaie Cardiovascular Medical and Research Center, Iran University of Medical Sciences, Tehran, Iran

    Mohsen Ghiasi

  3. Department of Cellular and Molecular Biology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

    Mohsen Ghiasi

  4. Chemical Injuries Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

    Hossein Aghamollaei

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  1. Mohamadsadegh Mohamadzadeh
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Contributions

All authors contributed to the concept and design of the study. Preparation, implementation, data collection, and analysis were done by Mohamadsadegh Mohamadzadeh, Mohsen Ghiasi, and Hossein Aghamollaei. The first draft of the manuscript was written by Mohamadsadegh Mohamadzadeh and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Hossein Aghamollaei.

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Communicated by Yusuf Akhter.

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Mohamadzadeh, M., Ghiasi, M. & Aghamollaei, H. Optimization of plasmid electrotransformation into Bacillus subtilis using an antibacterial peptide. Arch Microbiol 206, 116 (2024). https://doi.org/10.1007/s00203-024-03847-2

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  • DOI: https://doi.org/10.1007/s00203-024-03847-2

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