Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants
Food-borne illnesses caused by pathogenic Escherichia coli strains, especially enterohaemorrhagic E. coli (EHEC), are a serious public health problem, as debilitating disease and even death from such food poisonings have been repeatedly reported. Colicin M (ColM), a non-antibiotic antimicrobial prot...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-10-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2023.1271757/full |
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| author | Nataliia Shcherbak Heike Prochaska Kateryna Lystvan Yelizaveta Prokhorova Anatoli Giritch Mykola Kuchuk |
| author_facet | Nataliia Shcherbak Heike Prochaska Kateryna Lystvan Yelizaveta Prokhorova Anatoli Giritch Mykola Kuchuk |
| author_sort | Nataliia Shcherbak |
| collection | DOAJ |
| description | Food-borne illnesses caused by pathogenic Escherichia coli strains, especially enterohaemorrhagic E. coli (EHEC), are a serious public health problem, as debilitating disease and even death from such food poisonings have been repeatedly reported. Colicin M (ColM), a non-antibiotic antimicrobial protein produced by some strains of E. coli, has shown promising activity in controlling multiple enteropathogenic strains of E. coli and related pathogens. As contaminated green leafy vegetables are a frequent source of pathogenic E. coli infections, we genetically modified (GM) two edible crops, lettuce (Lactuca sativa L.) and mizuna (Brassica rapa subsp. nipposinica var. laciniata), to stably express the ColM gene and assessed the antibacterial activity of tissue extracts from these plants against selected E. coli strains in vitro. Transgenic plants of these species were developed using Agrobacterium-mediated transformation with a vector containing the ColM-coding gene (cma) under the control of the 35S promoter. Western blot analysis of recombinant ColM protein was performed in selected transgenic plants to confirm cma gene expression and quantify ColM accumulation. Extracts of transgenic plants expressing ColM showed significant activity against two major strains of EHEC (O157:H7 and O104:H4) as well as E. coli strains resistant to beta-lactam- and carbapenem-class antibiotics. Importantly, the antibacterial activity persisted in several subsequent generations of transgenic lettuce and mizuna plants that stably expressed the ColM gene. In addition, our results also show that the antibacterial activity of dried (up to 40°C) biomass of transgenic plants remained stable without a decrease for at least three months. |
| first_indexed | 2024-03-11T16:33:43Z |
| format | Article |
| id | doaj.art-f4ecd363f95f4d1487ad8074634187dc |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-03-11T16:33:43Z |
| publishDate | 2023-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-f4ecd363f95f4d1487ad8074634187dc2023-10-23T20:59:10ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2023-10-011410.3389/fpls.2023.12717571271757Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plantsNataliia Shcherbak0Heike Prochaska1Kateryna Lystvan2Yelizaveta Prokhorova3Anatoli Giritch4Mykola Kuchuk5Department of Genetic Engineering, Institute of Cell Biology and Genetic Engineering of National Academy of Sciences (NAS) of Ukraine, Kyiv, UkraineNomad Bioscience GmbH, Halle (Saale), GermanyDepartment of Genetic Engineering, Institute of Cell Biology and Genetic Engineering of National Academy of Sciences (NAS) of Ukraine, Kyiv, UkraineDepartment of Genetic Engineering, Institute of Cell Biology and Genetic Engineering of National Academy of Sciences (NAS) of Ukraine, Kyiv, UkraineNomad Bioscience GmbH, Halle (Saale), GermanyDepartment of Genetic Engineering, Institute of Cell Biology and Genetic Engineering of National Academy of Sciences (NAS) of Ukraine, Kyiv, UkraineFood-borne illnesses caused by pathogenic Escherichia coli strains, especially enterohaemorrhagic E. coli (EHEC), are a serious public health problem, as debilitating disease and even death from such food poisonings have been repeatedly reported. Colicin M (ColM), a non-antibiotic antimicrobial protein produced by some strains of E. coli, has shown promising activity in controlling multiple enteropathogenic strains of E. coli and related pathogens. As contaminated green leafy vegetables are a frequent source of pathogenic E. coli infections, we genetically modified (GM) two edible crops, lettuce (Lactuca sativa L.) and mizuna (Brassica rapa subsp. nipposinica var. laciniata), to stably express the ColM gene and assessed the antibacterial activity of tissue extracts from these plants against selected E. coli strains in vitro. Transgenic plants of these species were developed using Agrobacterium-mediated transformation with a vector containing the ColM-coding gene (cma) under the control of the 35S promoter. Western blot analysis of recombinant ColM protein was performed in selected transgenic plants to confirm cma gene expression and quantify ColM accumulation. Extracts of transgenic plants expressing ColM showed significant activity against two major strains of EHEC (O157:H7 and O104:H4) as well as E. coli strains resistant to beta-lactam- and carbapenem-class antibiotics. Importantly, the antibacterial activity persisted in several subsequent generations of transgenic lettuce and mizuna plants that stably expressed the ColM gene. In addition, our results also show that the antibacterial activity of dried (up to 40°C) biomass of transgenic plants remained stable without a decrease for at least three months.https://www.frontiersin.org/articles/10.3389/fpls.2023.1271757/fullbacteriocinscolicin Mtransgenic lettucetransgenic mizunapathogenic Escherichia coliSTEC |
| spellingShingle | Nataliia Shcherbak Heike Prochaska Kateryna Lystvan Yelizaveta Prokhorova Anatoli Giritch Mykola Kuchuk Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants Frontiers in Plant Science bacteriocins colicin M transgenic lettuce transgenic mizuna pathogenic Escherichia coli STEC |
| title | Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants |
| title_full | Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants |
| title_fullStr | Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants |
| title_full_unstemmed | Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants |
| title_short | Accumulation of colicin M protein and its biological activity in transgenic lettuce and mizuna plants |
| title_sort | accumulation of colicin m protein and its biological activity in transgenic lettuce and mizuna plants |
| topic | bacteriocins colicin M transgenic lettuce transgenic mizuna pathogenic Escherichia coli STEC |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2023.1271757/full |
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