Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system
Abstract Background After 2.83% genome reduction in Lactococcus lactis NZ9000, a good candidate host for proteins production was obtained in our previous work. However, the gene deletion process was time consuming and laborious. Here, we proposed a convenient gene deletion method suitable for large-...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12934-019-1249-z |
| _version_ | 1818301297637457920 |
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| author | Fulu Liu Yating Zhang Wanjin Qiao Duolong Zhu Haijin Xu Per Erik Joakim Saris Mingqiang Qiao |
| author_facet | Fulu Liu Yating Zhang Wanjin Qiao Duolong Zhu Haijin Xu Per Erik Joakim Saris Mingqiang Qiao |
| author_sort | Fulu Liu |
| collection | DOAJ |
| description | Abstract Background After 2.83% genome reduction in Lactococcus lactis NZ9000, a good candidate host for proteins production was obtained in our previous work. However, the gene deletion process was time consuming and laborious. Here, we proposed a convenient gene deletion method suitable for large-scale genome reduction in L. lactis NZ9000. Results Plasmid pNZ5417 containing a visually selectable marker P nisZ -lacZ was constructed, which allowed more efficient and convenient screening of gene deletion mutants. Using this plasmid, two large nonessential DNA regions, L-4A and L-5A, accounting for 1.25% of the chromosome were deleted stepwise in L. lactis 9k-3. When compared with the parent strain, the mutant L. lactis 9k-5A showed better growth characteristics, transformability, carbon metabolic capacity, and amino acids biosynthesis. Conclusions Thus, this study provides a convenient and efficient system for large-scale genome deletion in L. lactis through application of visually selectable marker, which could be helpful for rapid genome streamlining and generation of restructured L. lactis strains that can be used as cell factories. |
| first_indexed | 2024-12-13T05:20:47Z |
| format | Article |
| id | doaj.art-6e1857a075404fe8b713d88fbe9b75c8 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-13T05:20:47Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-6e1857a075404fe8b713d88fbe9b75c82022-12-21T23:58:19ZengBMCMicrobial Cell Factories1475-28592019-11-0118111110.1186/s12934-019-1249-zRestructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening systemFulu Liu0Yating Zhang1Wanjin Qiao2Duolong Zhu3Haijin Xu4Per Erik Joakim Saris5Mingqiang Qiao6Key Laboratory of Molecular Microbiology and Technology, College of Life Sciences, Ministry of Education, Nankai UniversityKey Laboratory of Molecular Microbiology and Technology, College of Life Sciences, Ministry of Education, Nankai UniversityKey Laboratory of Molecular Microbiology and Technology, College of Life Sciences, Ministry of Education, Nankai UniversityDepartment of Molecular Medicine, Morsani College of Medicine, University of South FloridaKey Laboratory of Molecular Microbiology and Technology, College of Life Sciences, Ministry of Education, Nankai UniversityDepartment of Food and Environmental Sciences, University of HelsinkiKey Laboratory of Molecular Microbiology and Technology, College of Life Sciences, Ministry of Education, Nankai UniversityAbstract Background After 2.83% genome reduction in Lactococcus lactis NZ9000, a good candidate host for proteins production was obtained in our previous work. However, the gene deletion process was time consuming and laborious. Here, we proposed a convenient gene deletion method suitable for large-scale genome reduction in L. lactis NZ9000. Results Plasmid pNZ5417 containing a visually selectable marker P nisZ -lacZ was constructed, which allowed more efficient and convenient screening of gene deletion mutants. Using this plasmid, two large nonessential DNA regions, L-4A and L-5A, accounting for 1.25% of the chromosome were deleted stepwise in L. lactis 9k-3. When compared with the parent strain, the mutant L. lactis 9k-5A showed better growth characteristics, transformability, carbon metabolic capacity, and amino acids biosynthesis. Conclusions Thus, this study provides a convenient and efficient system for large-scale genome deletion in L. lactis through application of visually selectable marker, which could be helpful for rapid genome streamlining and generation of restructured L. lactis strains that can be used as cell factories.http://link.springer.com/article/10.1186/s12934-019-1249-zLactococcus lactisVisually selectable markerLarge-scale genome deletion |
| spellingShingle | Fulu Liu Yating Zhang Wanjin Qiao Duolong Zhu Haijin Xu Per Erik Joakim Saris Mingqiang Qiao Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system Microbial Cell Factories Lactococcus lactis Visually selectable marker Large-scale genome deletion |
| title | Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system |
| title_full | Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system |
| title_fullStr | Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system |
| title_full_unstemmed | Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system |
| title_short | Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system |
| title_sort | restructured lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system |
| topic | Lactococcus lactis Visually selectable marker Large-scale genome deletion |
| url | http://link.springer.com/article/10.1186/s12934-019-1249-z |
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