Cargando…
Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria
BACKGROUND: Salmonella is a common foodborne pathogen, which can cause intestinal diseases. In the last decades, the overuse of antibiotics has led to a pandemic of drug-resistant bacterial infections. To tackle the burden of antimicrobial resistant pathogens, it is necessary to develop new antimic...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396760/ https://www.ncbi.nlm.nih.gov/pubmed/35999567 http://dx.doi.org/10.1186/s12934-022-01894-2 |
_version_ | 1784771992710283264 |
---|---|
author | Zhang, Shuhang Chang, Yan Zhang, Qing Yuan, Yingbo Qi, Qingsheng Lu, Xuemei |
author_facet | Zhang, Shuhang Chang, Yan Zhang, Qing Yuan, Yingbo Qi, Qingsheng Lu, Xuemei |
author_sort | Zhang, Shuhang |
collection | PubMed |
description | BACKGROUND: Salmonella is a common foodborne pathogen, which can cause intestinal diseases. In the last decades, the overuse of antibiotics has led to a pandemic of drug-resistant bacterial infections. To tackle the burden of antimicrobial resistant pathogens, it is necessary to develop new antimicrobial drugs with novel modes of action. However, the research and development of antibiotics has encountered bottlenecks, scientific hurdles in the development process, as well as safety and cost challenges. Phages and phage endolysins are promising antibacterial agents that can be used as an alternative to antibiotics. In this context, the expression of endolysin derived from different phages through microbial cells as a chassis seems to be an attractive strategy. RESULTS: In this study, a new endolysin from the Salmonella phage XFII-1, named XFII, was screened and obtained. The endolysin yield exceeded 100 mg/mL by heterologous expression from E. coli BL21 and short induction. The endolysin XFII exhibited high bactericidal activity at a concentration of 0.5 μg/mL and reduced the OD(600) nm of EDTA-pretreated E. coli JM109 from 0.8 to 0.2 within 5 min. XFII exhibited good thermo-resistance, as it was very stable at different temperatures from 20 to 80℃. Its bactericidal activity could keep constant at 4 °C for 175 days. In addition, the endolysin was able to exert lytic activity in eutrophic conditions, including LB medium and rabbit serum, and the lytic activity was even increased by 13.8% in 10% serum matrices. XFII also showed bactericidal activity against many Gram-negative bacteria, including Salmonella, E. coli, Acinetobacter baumannii, and Klebsiella pneumoniae. Surprisingly, the combination of endolysin XFII and chitosan showed a strong synergy in lysing E. coli and Salmonella without EDTA-pretreatment, and the OD(600) nm of E. coli decreased from 0.88 to 0.58 within 10 min. CONCLUSIONS: The novel globular endolysin XFII was screened and successfully expressed in E. coli BL21. Endolysin XFII exhibits a broad lysis spectrum, a rapid and strong bactericidal activity, good stability at high temperatures and under eutrophic conditions. Combined with chitosan, XFII could spontaneously lyse Gram-negative bacteria without pretreatment. This work presented the first characterization of combining endolysin and chitosan in spontaneously lysing Gram-negative bacteria in vitro. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01894-2. |
format | Online Article Text |
id | pubmed-9396760 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-93967602022-08-24 Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria Zhang, Shuhang Chang, Yan Zhang, Qing Yuan, Yingbo Qi, Qingsheng Lu, Xuemei Microb Cell Fact Research BACKGROUND: Salmonella is a common foodborne pathogen, which can cause intestinal diseases. In the last decades, the overuse of antibiotics has led to a pandemic of drug-resistant bacterial infections. To tackle the burden of antimicrobial resistant pathogens, it is necessary to develop new antimicrobial drugs with novel modes of action. However, the research and development of antibiotics has encountered bottlenecks, scientific hurdles in the development process, as well as safety and cost challenges. Phages and phage endolysins are promising antibacterial agents that can be used as an alternative to antibiotics. In this context, the expression of endolysin derived from different phages through microbial cells as a chassis seems to be an attractive strategy. RESULTS: In this study, a new endolysin from the Salmonella phage XFII-1, named XFII, was screened and obtained. The endolysin yield exceeded 100 mg/mL by heterologous expression from E. coli BL21 and short induction. The endolysin XFII exhibited high bactericidal activity at a concentration of 0.5 μg/mL and reduced the OD(600) nm of EDTA-pretreated E. coli JM109 from 0.8 to 0.2 within 5 min. XFII exhibited good thermo-resistance, as it was very stable at different temperatures from 20 to 80℃. Its bactericidal activity could keep constant at 4 °C for 175 days. In addition, the endolysin was able to exert lytic activity in eutrophic conditions, including LB medium and rabbit serum, and the lytic activity was even increased by 13.8% in 10% serum matrices. XFII also showed bactericidal activity against many Gram-negative bacteria, including Salmonella, E. coli, Acinetobacter baumannii, and Klebsiella pneumoniae. Surprisingly, the combination of endolysin XFII and chitosan showed a strong synergy in lysing E. coli and Salmonella without EDTA-pretreatment, and the OD(600) nm of E. coli decreased from 0.88 to 0.58 within 10 min. CONCLUSIONS: The novel globular endolysin XFII was screened and successfully expressed in E. coli BL21. Endolysin XFII exhibits a broad lysis spectrum, a rapid and strong bactericidal activity, good stability at high temperatures and under eutrophic conditions. Combined with chitosan, XFII could spontaneously lyse Gram-negative bacteria without pretreatment. This work presented the first characterization of combining endolysin and chitosan in spontaneously lysing Gram-negative bacteria in vitro. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01894-2. BioMed Central 2022-08-23 /pmc/articles/PMC9396760/ /pubmed/35999567 http://dx.doi.org/10.1186/s12934-022-01894-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Zhang, Shuhang Chang, Yan Zhang, Qing Yuan, Yingbo Qi, Qingsheng Lu, Xuemei Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria |
title | Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria |
title_full | Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria |
title_fullStr | Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria |
title_full_unstemmed | Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria |
title_short | Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria |
title_sort | characterization of salmonella endolysin xfii produced by recombinant escherichia coli and its application combined with chitosan in lysing gram-negative bacteria |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396760/ https://www.ncbi.nlm.nih.gov/pubmed/35999567 http://dx.doi.org/10.1186/s12934-022-01894-2 |
work_keys_str_mv | AT zhangshuhang characterizationofsalmonellaendolysinxfiiproducedbyrecombinantescherichiacolianditsapplicationcombinedwithchitosaninlysinggramnegativebacteria AT changyan characterizationofsalmonellaendolysinxfiiproducedbyrecombinantescherichiacolianditsapplicationcombinedwithchitosaninlysinggramnegativebacteria AT zhangqing characterizationofsalmonellaendolysinxfiiproducedbyrecombinantescherichiacolianditsapplicationcombinedwithchitosaninlysinggramnegativebacteria AT yuanyingbo characterizationofsalmonellaendolysinxfiiproducedbyrecombinantescherichiacolianditsapplicationcombinedwithchitosaninlysinggramnegativebacteria AT qiqingsheng characterizationofsalmonellaendolysinxfiiproducedbyrecombinantescherichiacolianditsapplicationcombinedwithchitosaninlysinggramnegativebacteria AT luxuemei characterizationofsalmonellaendolysinxfiiproducedbyrecombinantescherichiacolianditsapplicationcombinedwithchitosaninlysinggramnegativebacteria |