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Lactococcus lactis DPC5598, a plasmid-free derivative of a commercial starter, provides a valuable alternative host for culture improvement studies

Trotter, M ; Ross, R.P ; Fitzgerald, G.F ; Coffey, A

Journal of applied microbiology, 2002-01, Vol.93 (1), p.134-143 [Peer Reviewed Journal]

Oxford UK: Blackwell Science Ltd

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Title:
Lactococcus lactis DPC5598, a plasmid-free derivative of a commercial starter, provides a valuable alternative host for culture improvement studies
Author: Trotter, M ; Ross, R.P ; Fitzgerald, G.F ; Coffey, A
Subjects: adsorption ; Bacteriophages ; Biological and medical sciences ; Biotechnology ; Cheddar cheese ; Cheese - microbiology ; Chromosomes, Bacterial ; complement ; DNA Restriction Enzymes - genetics ; DNA, Bacterial - analysis ; Electrophoresis, Gel, Pulsed-Field ; fermentation ; Fermentation - genetics ; Fundamental and applied biological sciences. Psychology ; Genetic engineering ; Genetic technics ; genome ; Genome, Bacterial ; Lactococcus lactis ; Lactococcus lactis - genetics ; Lactococcus lactis - growth & development ; Lactococcus lactis - metabolism ; lactose ; Lactose - metabolism ; manufacturing ; Methods. Procedures. Technologies ; Miscellaneous ; Molecular Sequence Data ; Phenotype ; plasmid curing ; Plasmids ; proteinases ; pulsed-field gel electrophoresis ; starter cultures ; Synthetic digonucleotides and genes. Sequencing ; temperature
Is Part Of: Journal of applied microbiology, 2002-01, Vol.93 (1), p.134-143
Notes: ObjectType-Article-2
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Description: Aims: To generate a plasmid-free derivative of an extensively used industrial starter strain Lactococcus lactis DPC4268, which could be used as a backbone strain for starter improvement programmes. Methods and Results: DPC4268 containing four large plasmids was subjected to high temperature plasmid curing resulting in derivatives, each with a different plasmid complement of one, two or three different plasmids in addition to a plasmid-free derivative. Industrially relevant phenotypes were assigned to each plasmid on the basis of detailed phenotypic and genetic analyses and these were (a) proteinase activity (Prt, 60 kb) (b) lactose fermentation (Lac, 55 kb) (c) bacteriophage adsorption inhibition (Ads, 44 kb) and (d) type I restriction/modification (R/M, 40 kb). The plasmid-free variant of DPC4268 was shown to be transformable at frequencies comparable to the common laboratory strain L. lactis MG1614. Furthermore its genome was demonstrated to be significantly different from the laboratory strains L. lactis MG1614 and the recently sequenced L. lactis IL1403 genomes by pulsed-field gel electrophoresis. Conclusions: This study produced an easily transformable plasmid-free derivative which was genomically different from both MG1614 and IL1403. In addition, important plasmid-borne industrial traits, including two phage-resistance mechanisms, were identified in DPC4268. Significance and Impact of the Study: L. DPC4268 is a vitally important commercial strain used in the manufacture of Cheddar cheese. The generation of a plasmid-free derivative may provide an important backbone strain as a basis for future strain improvement purposes.
Publisher: Oxford UK: Blackwell Science Ltd
Language: English

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Lactococcus lactis DPC5598, a plasmid-free derivative of a commercial starter, provides a valuable alternative host for culture improvement studies

Trotter, M ; Ross, R.P ; Fitzgerald, G.F ; Coffey, A

Oxford UK: Blackwell Science Ltd

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