Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system

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Liu , F , Zhang , Y , Qiao , W , Zhu , D , Xu , H , Saris , P E J & Qiao , M 2019 , ' Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system ' , Microbial Cell Factories , vol. 18 , no. 1 , 198 . https://doi.org/10.1186/s12934-019-1249-z

Title: Restructured Lactococcus lactis strains with emergent properties constructed by a novel highly efficient screening system
Author: Liu, Fulu; Zhang, Yating; Qiao, Wanjin; Zhu, Duolong; Xu, Haijin; Saris, Per Erik Joakim; Qiao, Mingqiang
Contributor: University of Helsinki, Department of Microbiology
Date: 2019-11-14
Language: eng
Number of pages: 11
Belongs to series: Microbial Cell Factories
ISSN: 1475-2859
URI: http://hdl.handle.net/10138/311926
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.
Subject: Lactococcus lactis
Visually selectable marker
Large-scale genome deletion
ESCHERICHIA-COLI
GENOME REDUCTION
GENE-EXPRESSION
PROTEIN
PLASMID
IDENTIFICATION
TRANSFORMATION
DELETIONS
BACTERIA
GROWTH
1183 Plant biology, microbiology, virology
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