Cloning and expression of the Lactococcus lactis subsp. Cremoris SK11 gene encoding an extracellular serine proteinase

Vos, Willem M. de; Vos, Pieter; Haard, Hans de; Boerrigter, I.J.

doi:10.1016/0378-1119(89)90477-0

Cited by 117 publications

(108 citation statements)

References 18 publications

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“…It grows poorly in milk as compared to typical dairy L. lactis strains. Notably, the introduction of a plasmid, pNZ521 (de Vos et al 1989), encoding the lactococcal extracellular protease (PrtP) in strain KF147 significantly enhanced growth in milk (Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

“…Strains were grown either in M17 medium (Merck) supplemented either with 0.5% lactose (LM17) or 0.5% glucose (GM17), or in reconstituted skimmed milk (RSM) (Promex Spray 1% skimmed milk powder; Friesland Foods Butter). The plasmid pNZ521 (de Vos et al 1989) was introduced into strain KF147 by electroporation (Wells et al 1993), and transformants were selected by plating on medium supplemented with 5 mg/mL chloramphenicol. KF147 and all evolved variants were grown at 30°C.…”

Section: Bacterial Strains and Dna Techniquesmentioning

confidence: 99%

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Microbial domestication signatures of Lactococcus lactis can be reproduced by experimental evolution

Bachmann¹,

Starrenburg²,

Molenaar³

et al. 2011

Genome Res.

153

121

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Experimental evolution is a powerful approach to unravel how selective forces shape microbial genotypes and phenotypes. To this date, the available examples focus on the adaptation to conditions specific to the laboratory. The lactic acid bacterium Lactococcus lactis naturally occurs on plants and in dairy environments, and it is proposed that dairy strains originate from the plant niche. Here we investigate the adaptation of a L. lactis strain isolated from a plant to a dairy niche by propagating it for 1000 generations in milk. Two out of three independently evolved strains displayed significantly increased acidification rates and biomass yields in milk. Genome resequencing, revealed six, seven, and 28 mutations in the three strains, including point mutations in loci related to amino acid biosynthesis and transport and in the gene encoding MutL, which is involved in DNA mismatch repair. Two strains lost a conjugative transposon containing genes important in the plant niche but dispensable in milk. A plasmid carrying an extracellular protease was introduced by transformation. Although improving growth rate and growth yield significantly, the plasmid was rapidly lost. Comparative transcriptome and phenotypic analyses confirmed that major physiological changes associated with improved growth in milk relate to nitrogen metabolism and the loss or down-regulation of several pathways involved in the utilization of complex plant polymers. Reproducing the transition from the plant to the dairy niche through experimental evolution revealed several genome, transcriptome, and phenotype signatures that resemble those seen in strains isolated from either niche.

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Section: Resultsmentioning

confidence: 99%

Section: Bacterial Strains and Dna Techniquesmentioning

confidence: 99%

Microbial domestication signatures of Lactococcus lactis can be reproduced by experimental evolution

Bachmann¹,

Starrenburg²,

Molenaar³

et al. 2011

Genome Res.

153

121

View full text Add to dashboard Cite

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“…Preparation and transformation of competent L. lactis cells were performed as described by De Vos et al (1989). Southern transfer and hybridizations were all performed according to standard procedures (Sambrook et al, 1989) using the ECL kit (Amersham).…”

Section: Methodsmentioning

confidence: 99%

Six putative two-component regulatory systems isolated from Lactococcus lactis subsp. cremoris MG1363 The GenBank accession numbers for the sequences of the six 2CSs and surrounding ORFs determined in this work are AF172649, AF176556, AF178425, AF172650, AF176557 and AF176809 for systems A–F, respectively.

et al. 2000

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The genetic elements specifying six putative two-component regulatory systems (2CSs) were identified on the chromosome of Lactococcus lactis MG1363. These 2CSs appear to represent distinct loci, each containing a histidine kinase and response-regulator-encoding gene pair. Transcriptional analysis of the six 2CSs was performed either by generating transcriptional fusions to a reporter gene or by primer extension. Two of the systems appeared to be expressed constitutively at a high level, whilst the remaining four exhibited growth-phase-dependent expression. Insertional mutagenesis established that the two constitutively expressed 2CSs are necessary for normal cell growth and/or survival. Mutational analysis of the remaining four systems revealed that they are implicated in susceptibility to extreme pH, osmotic or oxidative conditions, or the regulation of phosphatase activity in L. lactis.

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“…Large-scale isolation of E. coli plasmid DNA for nucleotide sequence analysis was performed using Qiagen columns, according to the instructions of the manufacturer (Qiagen Inc.). Isolation and transformation of L. lactis plasmid DNA were performed as previously described (De Vos et al, 1989).…”

Section: Dna Isolation and Manipulationmentioning

confidence: 99%

Molecular characterization of the plasmid‐encoded eps gene cluster essential for exopolysaccharide biosynthesis in Lactococcus lactis

Kranenburg

Marugg

Swam

et al. 1997

Molecular Microbiology

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Cloning and expression of the Lactococcus lactis subsp. Cremoris SK11 gene encoding an extracellular serine proteinase

Cited by 117 publications

References 18 publications

Microbial domestication signatures of Lactococcus lactis can be reproduced by experimental evolution

Microbial domestication signatures of Lactococcus lactis can be reproduced by experimental evolution

Molecular characterization of the plasmid‐encoded eps gene cluster essential for exopolysaccharide biosynthesis in Lactococcus lactis

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