Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin
Abstract Background Hepcidin is an antimicrobial peptide involved in innate immunity, and the addition of hepcidin or probiotics expressing hepcidin to feed can significantly enhance the disease resistance of farmed animals. Lactobacillus plantarum has the advantages of stable colonization in the an...
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BMC
2025-07-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-025-02778-x |
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| author | Cheng-long Zhang Guangtao Chen Xiang Shan Ji Jian Teng Zhi Lei Yao Chun Lei Hu Yan Zhao |
| author_facet | Cheng-long Zhang Guangtao Chen Xiang Shan Ji Jian Teng Zhi Lei Yao Chun Lei Hu Yan Zhao |
| author_sort | Cheng-long Zhang |
| collection | DOAJ |
| description | Abstract Background Hepcidin is an antimicrobial peptide involved in innate immunity, and the addition of hepcidin or probiotics expressing hepcidin to feed can significantly enhance the disease resistance of farmed animals. Lactobacillus plantarum has the advantages of stable colonization in the animal gut, promotion of intestinal health, and efficient expression of foreign proteins. Previously, we have used L. plantarum (LP37) as a feed additive to promote the growth of Micropterus salmoides and Oreochromis mossambicus. The aim of this study is to construct a recombinant LP37 expressing hepcidin and evaluate its role in improving antibacterial ability and growth promotion in M. salmoides. Results Based on the expression vector pSIP403, we used green fluorescent protein (GFP) as a reporter gene to compare the effects of four promoters on the expression levels of foreign proteins and selected the promoter TUFA with the highest expression level. By introducing the ribosomal binding site (RBS) and secretory signal peptide sequence (SP-PGH), we constructed a novel vector capable of efficiently expressing M. salmoides hepcidin (MsHep) extracellularly. Transformation of the novel vector into LP37 to obtain the recombinant strain: LP37-MsHep. This recombinant LP37-MsHep demonstrated considerable antibacterial activity against Aeromonas hydrophila and several other microbes. The dietary supplementation of recombinant LP37-MsHep for 60 days significantly enhanced the survival rates of M. salmoides and reduced tissue bacterial load in M. salmoides post infection with A. hydrophila, and regulated gene expression of FPN1, IL-10, TNF- $$\alpha$$ α , and MHC-II involved in iron metabolism and immunity. Similar to LP37, dietary supplementation of recombinant LP37-MsHep also significantly improved the final body weight (FBW), weight gain ratio (WGR), and specific growth rate (SGR) and reduced the feed coefficient rate (FCR). Conclusions Collectively, the recombinant LP37-MsHep is capable of significantly improving growth performance and disease resistance of M. salmoides, presenting it as a promising green feed additive in aquaculture with broad application potential. |
| format | Article |
| id | doaj-art-008083e61eb74bb0ad375f7dc20e4f66 |
| institution | Kabale University |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-008083e61eb74bb0ad375f7dc20e4f662025-08-20T03:42:03ZengBMCMicrobial Cell Factories1475-28592025-07-0124111410.1186/s12934-025-02778-xConstruction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidinCheng-long Zhang0Guangtao Chen1Xiang Shan Ji2Jian Teng3Zhi Lei Yao4Chun Lei Hu5Yan Zhao6Key Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityKey Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityKey Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityKey Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityKey Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityKey Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityKey Laboratory of Efficient Utilization of Non-grain Feed Resources (Co-construction by Ministry and Province) of Ministry of Agriculture and Rural Affairs, Shandong Agricultural UniversityAbstract Background Hepcidin is an antimicrobial peptide involved in innate immunity, and the addition of hepcidin or probiotics expressing hepcidin to feed can significantly enhance the disease resistance of farmed animals. Lactobacillus plantarum has the advantages of stable colonization in the animal gut, promotion of intestinal health, and efficient expression of foreign proteins. Previously, we have used L. plantarum (LP37) as a feed additive to promote the growth of Micropterus salmoides and Oreochromis mossambicus. The aim of this study is to construct a recombinant LP37 expressing hepcidin and evaluate its role in improving antibacterial ability and growth promotion in M. salmoides. Results Based on the expression vector pSIP403, we used green fluorescent protein (GFP) as a reporter gene to compare the effects of four promoters on the expression levels of foreign proteins and selected the promoter TUFA with the highest expression level. By introducing the ribosomal binding site (RBS) and secretory signal peptide sequence (SP-PGH), we constructed a novel vector capable of efficiently expressing M. salmoides hepcidin (MsHep) extracellularly. Transformation of the novel vector into LP37 to obtain the recombinant strain: LP37-MsHep. This recombinant LP37-MsHep demonstrated considerable antibacterial activity against Aeromonas hydrophila and several other microbes. The dietary supplementation of recombinant LP37-MsHep for 60 days significantly enhanced the survival rates of M. salmoides and reduced tissue bacterial load in M. salmoides post infection with A. hydrophila, and regulated gene expression of FPN1, IL-10, TNF- $$\alpha$$ α , and MHC-II involved in iron metabolism and immunity. Similar to LP37, dietary supplementation of recombinant LP37-MsHep also significantly improved the final body weight (FBW), weight gain ratio (WGR), and specific growth rate (SGR) and reduced the feed coefficient rate (FCR). Conclusions Collectively, the recombinant LP37-MsHep is capable of significantly improving growth performance and disease resistance of M. salmoides, presenting it as a promising green feed additive in aquaculture with broad application potential.https://doi.org/10.1186/s12934-025-02778-xMicropterus salmoidesAntimicrobial PeptideHepcidinLactobacillus plantarumRecombinant strain |
| spellingShingle | Cheng-long Zhang Guangtao Chen Xiang Shan Ji Jian Teng Zhi Lei Yao Chun Lei Hu Yan Zhao Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin Microbial Cell Factories Micropterus salmoides Antimicrobial Peptide Hepcidin Lactobacillus plantarum Recombinant strain |
| title | Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin |
| title_full | Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin |
| title_fullStr | Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin |
| title_full_unstemmed | Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin |
| title_short | Construction and evaluation of recombinant Lactobacillus plantarum expressing Micropterus salmoides hepcidin |
| title_sort | construction and evaluation of recombinant lactobacillus plantarum expressing micropterus salmoides hepcidin |
| topic | Micropterus salmoides Antimicrobial Peptide Hepcidin Lactobacillus plantarum Recombinant strain |
| url | https://doi.org/10.1186/s12934-025-02778-x |
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