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Evaluation of the Antimicrobial Efficacy of Epidermal Mucus Extract from Air-Breathing Fish (Channa punctatus) and Identification of the Peptides Serving as Immune Components

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Abstract

Fish and other members of aquatic habitats are in constant contact with the microbes that trigger their immune system to produce immune components along with mucus. Compounds like lysozymes, proteases, lectins and peroxidase were reported in many studies. The study aims to identify and characterize the immune components present in the epidermal mucus of Channa punctatus. For this, antibacterial and antifungal efficacy was tested first using agar well diffusion assay and MIC was measured. Maximum activity against bacteria’s E. coli, S. aureus and Fungus H. gramineum was observed. FESEM analysis was performed to reveal the mechanism of the antimicrobial activity. Physico–chemical stability of mucus was also checked by exposing to various parameters such as (temperature, pH, proteases and surfactants). Precipitated proteins of mucus were separated using 12% SDS-PAGE assay. The band was digested using trypsin digestion and digested peptides were subjected to LCMSMS analysis for identification and on characterization using MASCOT servers 2 proteins (Hemoglobin subunit beta-A-globin chain and Galactose specific lectins nattectin) with number of fragmented unique peptides were obtained. Insilico predictions of antibacterial, antifungal, antiviral and anticancerous properties of peptides were also determined to support the role of mucus in microbial defense and therapeutical applications using the servers AMPfun, DBAASP and APD3.

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Data Availability

The datasets are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors thank the Department of Bioengineering and Biotechnology BIT Mesra for the facilities. The authors also thank Dr. Abhijit Dutta, Ranchi University, for his valuable suggestions. Authors would like to acknowledge Dr. Kunal Mukhopadhyay and Dr. Manish Kumar, Department of Bioengineering and Biotechnology BIT Mesra, for their suggestions in proteomics studies. First author acknowledges the Ministry of Minority Affairs India (MoMA) for Maulana Azad National Fellowship Grant/Award Number: [82 − 27/ 2019 (SA-III)].

Funding

The study was funded by Minority Affairs India (MoMA) for Maulana Azad National Fellowship Grant/Award Number: [82 − 27/ 2019 (SA-III)].

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Authors and Affiliations

  1. Department of Bioengineering and Biotechnology, Birla Institute of Technology, Mesra, Ranchi, Jharkhand, 835215, India

    Ahmed Hussain & Shashwati Ghosh Sachan

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  1. Ahmed Hussain
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  2. Shashwati Ghosh Sachan
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Contributions

AH: conceptualization, data curation; formal analysis; funding acquisition; investigation; methodology; software; visualization; roles/writing—original draft; and writing—review & editing. SGS: conceptualization; resources; supervision; validation; visualization; and writing—review & editing.

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Correspondence to Shashwati Ghosh Sachan.

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Ethical Approval

The animal study was approved by Institutional Animal Ethics Committee (IAEC) (Ref No. 1972/PH/BIT/PROV/64/21/IAEC) Birla Institute of Technology Mesra, Ranchi.

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Hussain, A., Ghosh Sachan, S. Evaluation of the Antimicrobial Efficacy of Epidermal Mucus Extract from Air-Breathing Fish (Channa punctatus) and Identification of the Peptides Serving as Immune Components. Int J Pept Res Ther 30, 4 (2024). https://doi.org/10.1007/s10989-023-10581-8

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