Bashir, Aideh

Hansen, Lars H.

Heller, Knut J.

Knøchel, Susanne

Kot, Witold Piotr

Neve, Horst

Nielsen, Cecilie Lykke Marvig

Sørensen, Søren Johannes

Vogensen, Finn Kvist

English

Copenhagen University Research Information System

u n i ve r s i t y  o f  co pe n h ag e n  Københavns UniversitetSequence and comparative analysis of Lactococcus lactis c2 dairy bacteriphages withdifferent thermal inactivationVogensen, Finn Kvist; Nielsen, Cecilie Lykke Marvig; Bashir, Aideh; Kot, Witold Piotr;Hansen, Lars H.; Neve, Horst; Heller, Knut J.; Sørensen, Søren Johannes; Knøchel, SusannePublication date:2011Document versionEarly version, also known as pre-printCitation for published version (APA):Vogensen, F. K., Nielsen, C. L. M., Bashir, A., Kot, W. P., Hansen, L. H., Neve, H., ... Knøchel, S. (2011).Sequence and comparative analysis of Lactococcus lactis c2 dairy bacteriphages with different thermalinactivation. Poster session presented at LAB10, Egmond aan Zee, Netherlands.Download date: 02. Feb. 2020D E P A R T M E N T   O F   F O O D   S C I E N C EF A C U L T Y   O F   L I F E   S C I E N C E S U N I V E R S I T Y   O F   C O P E N H A G E NSequence and comparative analysis of Lactococcus lactis c2 dairy bacteriophages with different thermal inactivation Finn K. Vogensen1, Cecilie L. Marvig1, Bashir Aideh1, Witold Kot1, Lars H. Hansen2, Horst Neve3, Knut J. Heller3, Søren J. Sørensen2, Susanne Knøchel1.1D t t f F d S i F lt f Lif S i U i it f C hIntroductionThe prolate headed Lactococcus lactis c2 phages are among the 3 most common species isolated in the dairy industry. It was previously believed that the c2 phage species showed a very narrow heat inactivation spectrum, many inactivated by traditional pasteurization treatment of cheese milk. We recently characterized the thermal inactivation of eight c2 phages [Marvig et al. (2011) Int. Dairy J.  21, 556-560] and showed that c2 phages had an inactivation span (8-log reduction) of at least 10°C from 70°C to 80°Cepar men  o  oo  c ence, acu y o  e c ences, n vers y o  open agen.2Department Biology, Faculty of Natural Sciences, University of Copenhagen.3Department of Microbiology and Biotechnology, Max Rubner-Institut, Kiel.Comparative genomics of c2 phagesWe have recently sequenced five c2 phages using 454 Next Generation Sequencing Technology. The draft sequence was aligned in Figure 1 using the Artemis Comparison Tool. The published sequences of c2 and bIL67 were included in the comparison. The comparison showed that c2 phages like the 936 phages have conserved genomes.    .Comparative genomics of c2 phages with different thermal inactivationThe phages P220 and P684 had identical host range (data not shown) and highly similar DNA sequence as seen from Figure 1  and Figure 2B.  However, the two phages have different thermal inactivation as seen in Figure 2A. At 70°C for 5 min P220 was completely inactivated  (8 log reduction) while P684 only had a 4.5 log reduction.Is the late expressed L10 gene product involved in thermal stability?When the genomes of P220 and P684 are compared the main difference in the structural part of the genome is a 93 bp deletion in the l10 gene homologue (Figure 3A). This gives a 31 a.a. deletion (Figure 3B)Figure 1: Comparison of the putative ORF’s from the c2 phages c2, P951, P635, bIL67, P220, P684 and CHPC670 using the Artemis Comparison Tool. Increasing colour intensity indicates increasing homologies. Blue ring indicate region in l10 homologue with deletion in P684 compared to P220.ker 0 4 kerA.345678910Log reduction (log PFU mL-1 )65°C70°C75°CA BMarkP220P684MarkConclusion•Phages of the c2 species have a conserved genome structure (Figure 1).•Phage P220 and P684 have almost identical genome sequence but differences in          thermal inactivation (Figure 2). The L10 gene product has in phage c2 has been described as a protein located by 012P220 P684PhageFigure 2. A. Thermal inactivation of P220 and P684 for 5 min at given temperature. B. Restriction endunuclease digestion pattern of P220 and P684 with HinfI. Marker is 1 kb ladderFigure 3. Comparaison of part of l10 gene sequence (A) and corresponding a.a. sequence (B)  B.•The more stable P684 have a 93 bp deletion (31 a.a.) in the l10 gene homologue (Figure 3)•L10 has in phage c2 been localized to the tail tip [Lubbers et al. (1995)]•L10 may be involved in tail initiation complex and thereby control of phage stabilityAcknowledgement We thank Chr. Hansen A/S and Danisco for providing the phages CHPC670 and P951, respectivelyimmuno-gold labeling to the tip of the tail [Lubbers et al. (1995) Appl. Environ. Microbiol., 61, 4348-4356], and suggested to be the anti-receptor . However, later L15 was shown to be the anti-receptor [Stuer-Lauridsen et al. (2003) Virology, 309, 10-17].We speculate that L10 could be part of the tail initiator complex and as such have confound influence of stability of the phage particle. Further work is needed to support this hypothesis.

Sequence and comparative analysis of Lactococcus lactis c2 dairy bacteriphages with different thermal inactivation

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Sequence and comparative analysis of Lactococcus lactis c2 dairy bacteriphages with different thermal inactivation

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