Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i>
<i>Ciona</i> molecule against microbes-A24 (CiMAM) isolated from the marine chordate <i>Ciona intestinalis</i> is an antimicrobial peptide. To generate CiMAM-expressing transgenic <i>Bacillus subtilis</i>, we constructed a plasmid expressing recombinant CiMAM (rCi...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-02-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/19/2/111 |
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| author | Bing-Chang Lee Jui-Che Tsai Cheng-Yung Lin Chun-Wei Hung Jin-Chuan Sheu Huai-Jen Tsai |
| author_facet | Bing-Chang Lee Jui-Che Tsai Cheng-Yung Lin Chun-Wei Hung Jin-Chuan Sheu Huai-Jen Tsai |
| author_sort | Bing-Chang Lee |
| collection | DOAJ |
| description | <i>Ciona</i> molecule against microbes-A24 (CiMAM) isolated from the marine chordate <i>Ciona intestinalis</i> is an antimicrobial peptide. To generate CiMAM-expressing transgenic <i>Bacillus subtilis</i>, we constructed a plasmid expressing recombinant CiMAM (rCiMAM) and introduced it into <i>B. subtilis.</i> Transgenic strains C117 and C166 were selected since they were able to highly and stably express rCiMAM. We studied the bactericidal activity of pepsin-digested extracts from rCiMAM-expressing strains against freshwater and euryhaline pathogens that commonly occur in aquaculture ponds and found no difference from that of lactoferricin-expressing strains. The bactericidal activity of 1-μL aliquot from a total 5.5 mL extracted from 5 mL of cultured C117 (1.45 × 10<sup>8</sup> CFU·mL<sup>−1</sup>) and C166 (2.17 × 10<sup>8</sup> CFU·mL<sup>−1</sup>) against halophilic bacteria was equivalent to the efficacy of 57.06 and 32.35 ng of Tetracycline against <i>Vibrio natriegens</i>, 47.07 and 25.2 ng against <i>V. parahaemolyticus</i>, and 58.17 and 36.55 ng against <i>V. alginolyticus,</i> respectively, indicating higher bactericidal activity of pepsin-extracts from rCiMAM-containing strains against halophilic bacteria compared to that from lactoferricin-containing strains. Since the antibacterial activity of rCiMAM-expressing <i>B. subtilis</i> strains shows higher competence against halophilic pathogens compared to that against freshwater and euryhaline pathogens, these strains are promising candidates to protect marine fish and shellfish from halophilic bacterial infection. |
| first_indexed | 2024-03-09T00:55:59Z |
| format | Article |
| id | doaj.art-bf9f586435284d3c9372ffa32361a22c |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-09T00:55:59Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-bf9f586435284d3c9372ffa32361a22c2023-12-11T16:54:26ZengMDPI AGMarine Drugs1660-33972021-02-0119211110.3390/md19020111Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i>Bing-Chang Lee0Jui-Che Tsai1Cheng-Yung Lin2Chun-Wei Hung3Jin-Chuan Sheu4Huai-Jen Tsai5Institute of Biomedical Sciences, Mackay Medical College, New Taipei City 25245, TaiwanInstitute of Molecular and Cellular Biology, National Taiwan University, Taipei 10617, TaiwanInstitute of Biomedical Sciences, Mackay Medical College, New Taipei City 25245, TaiwanInstitute of Molecular and Cellular Biology, National Taiwan University, Taipei 10617, TaiwanLiver Disease Prevention and Treatment Research Foundation, Taipei 10047, TaiwanInstitute of Biomedical Sciences, Mackay Medical College, New Taipei City 25245, Taiwan<i>Ciona</i> molecule against microbes-A24 (CiMAM) isolated from the marine chordate <i>Ciona intestinalis</i> is an antimicrobial peptide. To generate CiMAM-expressing transgenic <i>Bacillus subtilis</i>, we constructed a plasmid expressing recombinant CiMAM (rCiMAM) and introduced it into <i>B. subtilis.</i> Transgenic strains C117 and C166 were selected since they were able to highly and stably express rCiMAM. We studied the bactericidal activity of pepsin-digested extracts from rCiMAM-expressing strains against freshwater and euryhaline pathogens that commonly occur in aquaculture ponds and found no difference from that of lactoferricin-expressing strains. The bactericidal activity of 1-μL aliquot from a total 5.5 mL extracted from 5 mL of cultured C117 (1.45 × 10<sup>8</sup> CFU·mL<sup>−1</sup>) and C166 (2.17 × 10<sup>8</sup> CFU·mL<sup>−1</sup>) against halophilic bacteria was equivalent to the efficacy of 57.06 and 32.35 ng of Tetracycline against <i>Vibrio natriegens</i>, 47.07 and 25.2 ng against <i>V. parahaemolyticus</i>, and 58.17 and 36.55 ng against <i>V. alginolyticus,</i> respectively, indicating higher bactericidal activity of pepsin-extracts from rCiMAM-containing strains against halophilic bacteria compared to that from lactoferricin-containing strains. Since the antibacterial activity of rCiMAM-expressing <i>B. subtilis</i> strains shows higher competence against halophilic pathogens compared to that against freshwater and euryhaline pathogens, these strains are promising candidates to protect marine fish and shellfish from halophilic bacterial infection.https://www.mdpi.com/1660-3397/19/2/111CiMAMantimicrobial peptidelactoferricintransgenic linebactericidal activity |
| spellingShingle | Bing-Chang Lee Jui-Che Tsai Cheng-Yung Lin Chun-Wei Hung Jin-Chuan Sheu Huai-Jen Tsai Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i> Marine Drugs CiMAM antimicrobial peptide lactoferricin transgenic line bactericidal activity |
| title | Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i> |
| title_full | Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i> |
| title_fullStr | Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i> |
| title_full_unstemmed | Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i> |
| title_short | Using <i>Bacillus subtilis</i> as a Host Cell to Express an Antimicrobial Peptide from the Marine Chordate <i>Ciona intestinalis</i> |
| title_sort | using i bacillus subtilis i as a host cell to express an antimicrobial peptide from the marine chordate i ciona intestinalis i |
| topic | CiMAM antimicrobial peptide lactoferricin transgenic line bactericidal activity |
| url | https://www.mdpi.com/1660-3397/19/2/111 |
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