Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity
Fungal species resistant to current antifungal agents are considered as a serious threat to human health, the dilemma that has dragged attentions toward other sources of antifungals such as antimicrobial peptides (AMPs). In order to improve biological activity of a recently described antifungal pept...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-04-01
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| Series: | Frontiers in Fungal Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/ffunb.2021.638595/full |
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| author | Sima Sadat Seyedjavadi Soghra Khani Jafar Amani Raheleh Halabian Mehdi Goudarzi Hamideh Mahmoodzadeh Hosseini Ali Eslamifar Masoomeh Shams-Ghahfarokhi Abbas Ali Imani Fooladi Mehdi Razzaghi-Abyaneh |
| author_facet | Sima Sadat Seyedjavadi Soghra Khani Jafar Amani Raheleh Halabian Mehdi Goudarzi Hamideh Mahmoodzadeh Hosseini Ali Eslamifar Masoomeh Shams-Ghahfarokhi Abbas Ali Imani Fooladi Mehdi Razzaghi-Abyaneh |
| author_sort | Sima Sadat Seyedjavadi |
| collection | DOAJ |
| description | Fungal species resistant to current antifungal agents are considered as a serious threat to human health, the dilemma that has dragged attentions toward other sources of antifungals such as antimicrobial peptides (AMPs). In order to improve biological activity of a recently described antifungal peptide MCh-AMP1 from Matricaria chamomilla flowers, MCh-AMP1dimer (DiMCh-AMP1), containing 61 amino acid residues connected by flexible linker (GPDGSGPDESGPDES), was designed and expressed in Escherichia coli, and its structure was analyzed using bioinformatics tools. DiMCh-AMP1 synthetic gene was cloned into pET-28a expression vector, which was then used to transform E. coli BL21 (DE3) strain. His-tag purification was achieved using metal-chelate affinity chromatography. Because there is no methionine residue in the DiMCh-AMP1 sequence, cyanogen bromide was successfully used to separate the target product from the tag. Reverse-phase high-performance liquid chromatography was used as the final step of purification. Results showed that recombinant peptide was produced in considerable amounts (0.9 mg/L) with improved antifungal activity toward both yeasts and molds compared to its monomeric counterpart. The minimum inhibition concentration and minimum fungicidal concentration values of DiMCh-AMP1 against Candida and Aspergillus species were reported in the range of 1.67–6.66 μM and 3.33–26.64 μM, respectively. Our results showed that while antifungal activity of dimerized peptide was improved considerably, its cytotoxicity was decreased, implying that DiMCh-AMP1 could be a potential candidate to design an effective antifungal agent against pathogenic yeasts and molds. |
| first_indexed | 2024-12-21T18:34:54Z |
| format | Article |
| id | doaj.art-a3404cea4e1e4efdaed823543aa690da |
| institution | Directory Open Access Journal |
| issn | 2673-6128 |
| language | English |
| last_indexed | 2024-12-21T18:34:54Z |
| publishDate | 2021-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Fungal Biology |
| spelling | doaj.art-a3404cea4e1e4efdaed823543aa690da2022-12-21T18:54:11ZengFrontiers Media S.A.Frontiers in Fungal Biology2673-61282021-04-01210.3389/ffunb.2021.638595638595Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and CytotoxicitySima Sadat Seyedjavadi0Soghra Khani1Jafar Amani2Raheleh Halabian3Mehdi Goudarzi4Hamideh Mahmoodzadeh Hosseini5Ali Eslamifar6Masoomeh Shams-Ghahfarokhi7Abbas Ali Imani Fooladi8Mehdi Razzaghi-Abyaneh9Department of Mycology, Pasteur Institute of Iran, Tehran, IranDepartment of Mycology, Pasteur Institute of Iran, Tehran, IranApplied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, IranApplied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, IranDepartment of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranApplied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, IranDepartment of Clinical Research, Pasteur Institute of Iran, Tehran, IranDepartment of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranApplied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, IranDepartment of Mycology, Pasteur Institute of Iran, Tehran, IranFungal species resistant to current antifungal agents are considered as a serious threat to human health, the dilemma that has dragged attentions toward other sources of antifungals such as antimicrobial peptides (AMPs). In order to improve biological activity of a recently described antifungal peptide MCh-AMP1 from Matricaria chamomilla flowers, MCh-AMP1dimer (DiMCh-AMP1), containing 61 amino acid residues connected by flexible linker (GPDGSGPDESGPDES), was designed and expressed in Escherichia coli, and its structure was analyzed using bioinformatics tools. DiMCh-AMP1 synthetic gene was cloned into pET-28a expression vector, which was then used to transform E. coli BL21 (DE3) strain. His-tag purification was achieved using metal-chelate affinity chromatography. Because there is no methionine residue in the DiMCh-AMP1 sequence, cyanogen bromide was successfully used to separate the target product from the tag. Reverse-phase high-performance liquid chromatography was used as the final step of purification. Results showed that recombinant peptide was produced in considerable amounts (0.9 mg/L) with improved antifungal activity toward both yeasts and molds compared to its monomeric counterpart. The minimum inhibition concentration and minimum fungicidal concentration values of DiMCh-AMP1 against Candida and Aspergillus species were reported in the range of 1.67–6.66 μM and 3.33–26.64 μM, respectively. Our results showed that while antifungal activity of dimerized peptide was improved considerably, its cytotoxicity was decreased, implying that DiMCh-AMP1 could be a potential candidate to design an effective antifungal agent against pathogenic yeasts and molds.https://www.frontiersin.org/articles/10.3389/ffunb.2021.638595/fullantifungal activitycytotoxicityantimicrobial peptideAspergillusCandidadimeric antifungal peptides |
| spellingShingle | Sima Sadat Seyedjavadi Soghra Khani Jafar Amani Raheleh Halabian Mehdi Goudarzi Hamideh Mahmoodzadeh Hosseini Ali Eslamifar Masoomeh Shams-Ghahfarokhi Abbas Ali Imani Fooladi Mehdi Razzaghi-Abyaneh Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity Frontiers in Fungal Biology antifungal activity cytotoxicity antimicrobial peptide Aspergillus Candida dimeric antifungal peptides |
| title | Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity |
| title_full | Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity |
| title_fullStr | Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity |
| title_full_unstemmed | Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity |
| title_short | Design, Dimerization, and Recombinant Production of MCh-AMP1–Derived Peptide in Escherichia coli and Evaluation of Its Antifungal Activity and Cytotoxicity |
| title_sort | design dimerization and recombinant production of mch amp1 derived peptide in escherichia coli and evaluation of its antifungal activity and cytotoxicity |
| topic | antifungal activity cytotoxicity antimicrobial peptide Aspergillus Candida dimeric antifungal peptides |
| url | https://www.frontiersin.org/articles/10.3389/ffunb.2021.638595/full |
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