A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains
Lactoferrin (LF) is a multifunctional protein found in mammals, and it shows broad-spectrum antimicrobial activity. To improve the functional properties of specific probiotics in order to provide both the beneficial characteristics of lactic acid bacteria and the biological activity of LF, cDNAs of...
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MDPI AG
2021-01-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/10/2/120 |
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| author | Zhen-Shu Liu Chuen-Fu Lin Chung-Pei Lee Min-Chi Hsieh Hung-Fu Lu Ying-Fang Chen Yu-We Ku Po-Wen Chen |
| author_facet | Zhen-Shu Liu Chuen-Fu Lin Chung-Pei Lee Min-Chi Hsieh Hung-Fu Lu Ying-Fang Chen Yu-We Ku Po-Wen Chen |
| author_sort | Zhen-Shu Liu |
| collection | DOAJ |
| description | Lactoferrin (LF) is a multifunctional protein found in mammals, and it shows broad-spectrum antimicrobial activity. To improve the functional properties of specific probiotics in order to provide both the beneficial characteristics of lactic acid bacteria and the biological activity of LF, cDNAs of bovine LF (BLF), human LF (HLF), or porcine LF (PLF) were cloned into a nisin-inducible plasmid. These were then transformed into the selected eight probiotics, which are LF-resistant hosts. Expression of recombinant LFs (rLFs) was analyzed via SDS-PAGE and Western blot analysis. Although the selected host strains may not contain the <i>nisRK</i> genes (NisK, the sensor kinase; NisR, the regulator protein), the components of autoregulation, a low level of LFs expression can be successfully induced by using nisin within bacterial cells in a time-dependent manner in three engineered clones, including <i>Lactobacillus delbrueckii</i>/HLF, <i>L. delbrueckii</i>/BLF, and <i>L. gasseri</i>/BLF. <i>Lactobacillus delbrueckii</i> and <i>Lactobacillus gasseri</i> originate from yogurt and human milk, respectively, and both strains are functional probiotic strains. Therefore, we further compared the antibacterial activities of disrupted recombinant probiotic clones, conventional strains (host control), and vector control ones by using agar diffusion and broth inhibition analysis, and the expression of rLFs in the above three clones considerately improved their antibacterial efficacies against four important food-borne pathogens, namely, <i>Escherichia coli</i>, <i>Staphylococcus aureus</i>, <i>Enterococcus faecalis</i>, and <i>Salmonella</i><i>enterica</i>. In conclusion, this study provides a simple strategy for the production of functional LFs (BLF and HLF) in both functional and LF-resistant hosts for applications in the field. |
| first_indexed | 2024-03-09T03:34:34Z |
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| id | doaj.art-684aecfafe1649e2958c5f4083114867 |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-09T03:34:34Z |
| publishDate | 2021-01-01 |
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| series | Antibiotics |
| spelling | doaj.art-684aecfafe1649e2958c5f40831148672023-12-03T14:50:48ZengMDPI AGAntibiotics2079-63822021-01-0110212010.3390/antibiotics10020120A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic StrainsZhen-Shu Liu0Chuen-Fu Lin1Chung-Pei Lee2Min-Chi Hsieh3Hung-Fu Lu4Ying-Fang Chen5Yu-We Ku6Po-Wen Chen7Department of Safety, Health and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, TaiwanDepartment of Veterinary Medicine, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 91201, TaiwanSchool of Nursing, National Taipei University of Nursing and Health Sciences, Taipei 11219, TaiwanDepartment of Safety, Health and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, TaiwanDepartment of Safety, Health and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, TaiwanDepartment of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanDepartment of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanDepartment of Veterinary Medicine, National Chung Hsing University, Taichung 40249, TaiwanLactoferrin (LF) is a multifunctional protein found in mammals, and it shows broad-spectrum antimicrobial activity. To improve the functional properties of specific probiotics in order to provide both the beneficial characteristics of lactic acid bacteria and the biological activity of LF, cDNAs of bovine LF (BLF), human LF (HLF), or porcine LF (PLF) were cloned into a nisin-inducible plasmid. These were then transformed into the selected eight probiotics, which are LF-resistant hosts. Expression of recombinant LFs (rLFs) was analyzed via SDS-PAGE and Western blot analysis. Although the selected host strains may not contain the <i>nisRK</i> genes (NisK, the sensor kinase; NisR, the regulator protein), the components of autoregulation, a low level of LFs expression can be successfully induced by using nisin within bacterial cells in a time-dependent manner in three engineered clones, including <i>Lactobacillus delbrueckii</i>/HLF, <i>L. delbrueckii</i>/BLF, and <i>L. gasseri</i>/BLF. <i>Lactobacillus delbrueckii</i> and <i>Lactobacillus gasseri</i> originate from yogurt and human milk, respectively, and both strains are functional probiotic strains. Therefore, we further compared the antibacterial activities of disrupted recombinant probiotic clones, conventional strains (host control), and vector control ones by using agar diffusion and broth inhibition analysis, and the expression of rLFs in the above three clones considerately improved their antibacterial efficacies against four important food-borne pathogens, namely, <i>Escherichia coli</i>, <i>Staphylococcus aureus</i>, <i>Enterococcus faecalis</i>, and <i>Salmonella</i><i>enterica</i>. In conclusion, this study provides a simple strategy for the production of functional LFs (BLF and HLF) in both functional and LF-resistant hosts for applications in the field.https://www.mdpi.com/2079-6382/10/2/120recombinant lactoferrinprobioticantibacterial activityfood-borne pathogen |
| spellingShingle | Zhen-Shu Liu Chuen-Fu Lin Chung-Pei Lee Min-Chi Hsieh Hung-Fu Lu Ying-Fang Chen Yu-We Ku Po-Wen Chen A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains Antibiotics recombinant lactoferrin probiotic antibacterial activity food-borne pathogen |
| title | A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains |
| title_full | A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains |
| title_fullStr | A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains |
| title_full_unstemmed | A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains |
| title_short | A Single Plasmid of Nisin-Controlled Bovine and Human Lactoferrin Expressing Elevated Antibacterial Activity of Lactoferrin-Resistant Probiotic Strains |
| title_sort | single plasmid of nisin controlled bovine and human lactoferrin expressing elevated antibacterial activity of lactoferrin resistant probiotic strains |
| topic | recombinant lactoferrin probiotic antibacterial activity food-borne pathogen |
| url | https://www.mdpi.com/2079-6382/10/2/120 |
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