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.2010 Jul 23;5(7):e11748.
doi: 10.1371/journal.pone.0011748.

The complete genome of Propionibacterium freudenreichii CIRM-BIA1, a hardy actinobacterium with food and probiotic applications

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The complete genome of Propionibacterium freudenreichii CIRM-BIA1, a hardy actinobacterium with food and probiotic applications

Hélène Falentin et al. PLoS One..

Abstract

Background: Propionibacterium freudenreichii is essential as a ripening culture in Swiss-type cheeses and is also considered for its probiotic use. This species exhibits slow growth, low nutritional requirements, and hardiness in many habitats. It belongs to the taxonomic group of dairy propionibacteria, in contrast to the cutaneous species P. acnes. The genome of the type strain, P. freudenreichii subsp. shermanii CIRM-BIA1 (CIP 103027(T)), was sequenced with an 11-fold coverage.

Methodology/principal findings: The circular chromosome of 2.7 Mb of the CIRM-BIA1 strain has a GC-content of 67% and contains 22 different insertion sequences (3.5% of the genome in base pairs). Using a proteomic approach, 490 of the 2439 predicted proteins were confirmed. The annotation revealed the genetic basis for the hardiness of P. freudenreichii, as the bacterium possesses a complete enzymatic arsenal for de novo biosynthesis of aminoacids and vitamins (except panthotenate and biotin) as well as sequences involved in metabolism of various carbon sources, immunity against phages, duplicated chaperone genes and, interestingly, genes involved in the management of polyphosphate, glycogen and trehalose storage. The complete biosynthesis pathway for a bifidogenic compound is described, as well as a high number of surface proteins involved in interactions with the host and present in other probiotic bacteria. By comparative genomics, no pathogenicity factors found in P. acnes or in other pathogenic microbial species were identified in P. freudenreichii, which is consistent with the Generally Recognized As Safe and Qualified Presumption of Safety status of P. freudenreichii. Various pathways for formation of cheese flavor compounds were identified: the Wood-Werkman cycle for propionic acid formation, amino acid degradation pathways resulting in the formation of volatile branched chain fatty acids, and esterases involved in the formation of free fatty acids and esters.

Conclusions/significance: With the exception of its ability to degrade lactose, P. freudenreichii seems poorly adapted to dairy niches. This genome annotation opens up new prospects for the understanding of the P. freudenreichii probiotic activity.

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Conflict of interest statement

Competing Interests:The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Circular representation of the genome ofP. freudenreichii strain CIRM-BIA1T.
The outer circle shows the coding sequence by functional category: cell envelope in green, metabolism in blue, replication, transcription and traduction in yellow, other functions in orange, hypothetical proteins in violet. The second circle shows red blocks for coding sequences on the (+) strand. The third circle shows blue blocks for coding sequences on the (−) strand. The fourth circle shows rRNA in purple and tRNA in light purple. The fifth circle shows the insertion sequences and phage integrase in green. The sixth circle shows the precentage GC (window size 10,000 bp, step size 2000 bp) ranging from 59 to 72% with a step of 1%. The seventh circle shows the GC skew (window size 10,000 bp, step size 2000 bp) ranging from −0.13 to 0.09 with a step of 0.01%. The replication origin is between 10,426 bp and 12,300 bp.
Figure 2
Figure 2. InP. freudenreichii strain CIRM-BIA1T, genes involved in lactose metabolism are bordered by a transposase and integrase.
The lactose locus consists of three genes, PFREUD_02370, PFREUD_02360 and PFREUD_02350 (numbers are indicated below the arrows), which encode a βgalactosidase, LacZ, a galactoside transporter, and an UDP-glucose isomerase, GalE, respectively.Lac genes may have been acquired through a horizontal transfer event. PFREUD_02380 encodes the N-terminal end of an N-acetylmuramic acid 6-phosphate etherase and PFREUD_02390 encodes a transcriptional regulator.
Figure 3
Figure 3. Homology dot plot comparingP. freudenreichii strain CIRM-BIA1T andP. acnes at the protein level.
dnaA is at the zero position. Forward matches are displayed in red and reverse matches are displayed in blue. Relatively high synteny along the genome is observed, with the exception of two inversions between 90,000–120,000 bp and 100,000–115,000 bp.
Figure 4
Figure 4. Vitamin B12 biosynthesis inP. freudenreichii strain CIRM-BIA1T.
(A) Vitamin B12 pathway: enzyme number (red), gene name (green) and locus tag (black). (B) Four loci (a, b, c, d) are involved in vitamin B12 biosynthesis. Gene names are indicated above the arrows, locus tags (PFREUD) are indicated below the arrows. The locus (a) codes for the cobalt ABC transporter. The colors of the arrows used in (B) for locus b, c and d are also used for the pathway background in (A), with the exeption of the steps in the white background corresponding to isolated genes (hemD andcobT2/bluB).
Figure 5
Figure 5. Schematic representation of the Wood-Werkman cycle (in blue) and tricarboxylic acid (TCA) cycle (in black) inP. freudenreichii strain CIRM-BIA1T.
Enzyme numbers are in red, gene names are in green, and locus tags are in black. Reactions are directed toward propionate production, but are reversible.
See this image and copyright information in PMC

References

    1. Cousin FJ, Mater DD, Foligné B, Jan G. Dairy propionibacteria as human probiotics: A review of recent evidence (in press). Dairy Sci Technol. 2010. doi:10.1051/dst/2010032. - DOI
    1. von Freudenreich E, Orla-Jensen O. Uber die in Emmentalerkäse stattfindene Propionsäure-gärung. Zentralbl Bakteriol. 1906;17:529–546.
    1. Cummins CS, Johnson JL. The genus Propionibacterium. In: Balows E, Truper HG, Dworkin M, Harder W, Schleifer KH, editors. The prokaryotes. Springer Verlag; 1992.
    1. Thierry A, Maillard MB. Production of cheese flavour compounds derived from amino acid catabolism by Propionibacterium freudenreichii. Lait. 2002;82:17–32.
    1. Thierry A, Maillard MB, Richoux R, Kerjean JR, Lortal S. Propionibacterium freudenreichii strains quantitatively affect production of volatile compounds in Swiss cheese. Lait. 2005;85:57–74.

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