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Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin

BACKGROUND: Lactobacillus casei (L. casei) is well known for its probiotic property in human and animals. Lactoferricin (Lfcin) polypeptide can effectively modulate host immune responses and have antimicrobial activity in vivo and in vitro. In order to develop a food-grade L. casei system constituti...

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Autores principales: Zhou, Han, Li, Xuechun, Wang, Zongying, Yin, Jiyuan, Tan, Hongchao, Wang, Li, Qiao, Xinyuan, Jiang, Yanping, Cui, Wen, Liu, Min, Li, Yijing, Xu, Yigang, Tang, Lijie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6020375/
https://www.ncbi.nlm.nih.gov/pubmed/29945678
http://dx.doi.org/10.1186/s12917-018-1516-y
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author Zhou, Han
Li, Xuechun
Wang, Zongying
Yin, Jiyuan
Tan, Hongchao
Wang, Li
Qiao, Xinyuan
Jiang, Yanping
Cui, Wen
Liu, Min
Li, Yijing
Xu, Yigang
Tang, Lijie
author_facet Zhou, Han
Li, Xuechun
Wang, Zongying
Yin, Jiyuan
Tan, Hongchao
Wang, Li
Qiao, Xinyuan
Jiang, Yanping
Cui, Wen
Liu, Min
Li, Yijing
Xu, Yigang
Tang, Lijie
author_sort Zhou, Han
collection PubMed
description BACKGROUND: Lactobacillus casei (L. casei) is well known for its probiotic property in human and animals. Lactoferricin (Lfcin) polypeptide can effectively modulate host immune responses and have antimicrobial activity in vivo and in vitro. In order to develop a food-grade L. casei system constitutively expressing bovine Lfcin, this study constructed a thymidine auxotrophy (ΔthyA) recombinant L. casei. RESULTS: Based on the thymidylate synthase gene (thyA) insert site, LFEC(Lfcin expression cassette)was inserted into L. casei genome through homologous recombination, successfully expressed and could be stably inherited. The recombinant L. casei, ΔthyA L. casei-LFEC, is sensitive to chloramphenicol and limited when cultured without thymine. Meanwhile, ΔthyA L. casei-LFEC has both good antibacterial activity against Escherichia coli and Staphylococcus aureus and antiviral activity against porcine epidemic diarrhea virus (PEDV). CONCLUSIONS: We successfully constructed a recombinant L. casei strain expressing Lfcin, ΔthyA L. casei-LFEC, which could only survive in the presence of thymine, and had excellent antimicrobial and antiviral activity with good genetic stability and sensitivity. This research provides a cost-effective alternative to the antibiotics with additional biological functions and wider applicability prospect. Using ΔthyA as the selectable mark instead of antibiotic to construct genetic engineering L.casei provides a safe and effective approach of feed additives in livestock raising.
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spelling pubmed-60203752018-07-06 Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin Zhou, Han Li, Xuechun Wang, Zongying Yin, Jiyuan Tan, Hongchao Wang, Li Qiao, Xinyuan Jiang, Yanping Cui, Wen Liu, Min Li, Yijing Xu, Yigang Tang, Lijie BMC Vet Res Research Article BACKGROUND: Lactobacillus casei (L. casei) is well known for its probiotic property in human and animals. Lactoferricin (Lfcin) polypeptide can effectively modulate host immune responses and have antimicrobial activity in vivo and in vitro. In order to develop a food-grade L. casei system constitutively expressing bovine Lfcin, this study constructed a thymidine auxotrophy (ΔthyA) recombinant L. casei. RESULTS: Based on the thymidylate synthase gene (thyA) insert site, LFEC(Lfcin expression cassette)was inserted into L. casei genome through homologous recombination, successfully expressed and could be stably inherited. The recombinant L. casei, ΔthyA L. casei-LFEC, is sensitive to chloramphenicol and limited when cultured without thymine. Meanwhile, ΔthyA L. casei-LFEC has both good antibacterial activity against Escherichia coli and Staphylococcus aureus and antiviral activity against porcine epidemic diarrhea virus (PEDV). CONCLUSIONS: We successfully constructed a recombinant L. casei strain expressing Lfcin, ΔthyA L. casei-LFEC, which could only survive in the presence of thymine, and had excellent antimicrobial and antiviral activity with good genetic stability and sensitivity. This research provides a cost-effective alternative to the antibiotics with additional biological functions and wider applicability prospect. Using ΔthyA as the selectable mark instead of antibiotic to construct genetic engineering L.casei provides a safe and effective approach of feed additives in livestock raising. BioMed Central 2018-06-27 /pmc/articles/PMC6020375/ /pubmed/29945678 http://dx.doi.org/10.1186/s12917-018-1516-y Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Zhou, Han
Li, Xuechun
Wang, Zongying
Yin, Jiyuan
Tan, Hongchao
Wang, Li
Qiao, Xinyuan
Jiang, Yanping
Cui, Wen
Liu, Min
Li, Yijing
Xu, Yigang
Tang, Lijie
Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin
title Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin
title_full Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin
title_fullStr Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin
title_full_unstemmed Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin
title_short Construction and characterization of thymidine auxotrophic (ΔthyA) recombinant Lactobacillus casei expressing bovine lactoferricin
title_sort construction and characterization of thymidine auxotrophic (δthya) recombinant lactobacillus casei expressing bovine lactoferricin
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6020375/
https://www.ncbi.nlm.nih.gov/pubmed/29945678
http://dx.doi.org/10.1186/s12917-018-1516-y
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