A Temporal -omic Study of Propionibacterium freudenreichii CIRM-BIA1T Adaptation Strategies in Conditions Mimicking Cheese Ripening in the Cold

Propionibacterium freudenreichii is used as a ripening culture in Swiss cheese manufacture. It grows when cheeses are ripened in a warm room (about 24°C). Cheeses with an acceptable eye formation level are transferred to a cold room (about 4°C), inducing a marked slowdown of propionic fermentation, but P. freudenreichii remains active in the cold. To investigate the P. freudenreichii strategies of adaptation and survival in the cold, we performed the first global gene expression profile for this species. The time-course transcriptomic response of P. freudenreichii CIRM-BIA1T strain was analyzed at five times of incubation, during growth at 30°C then for 9 days at 4°C, under conditions preventing nutrient starvation. Gene expression was also confirmed by RT-qPCR for 28 genes. In addition, proteomic experiments were carried out and the main metabolites were quantified. Microarray analysis revealed that 565 genes (25% of the protein-coding sequences of P. freudenreichii genome) were differentially expressed during transition from 30°C to 4°C (P<0.05 and |fold change|>1). At 4°C, a general slowing down was observed for genes implicated in the cell machinery. On the contrary, P. freudenreichii CIRM-BIA1T strain over-expressed genes involved in lactate, alanine and serine conversion to pyruvate, in gluconeogenesis, and in glycogen synthesis. Interestingly, the expression of different genes involved in the formation of important cheese flavor compounds, remained unchanged at 4°C. This could explain the contribution of P. freudenreichii to cheese ripening even in the cold. In conclusion, P. freudenreichii remains metabolically active at 4°C and induces pathways to maintain its long-term survival

The Complete Genome of Propionibacterium freudenreichii CIRM-BIA1T, a Hardy Actinobacterium with Food and Probiotic Applications

Background: Propionibacterium freudenreichii is essential as a ripening culture in Swiss-type cheeses and is also considered for its probiotic use [1]. 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 103027T), 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

Identification of a glucan synthase gene potentially coding for exopolysaccharide production by Propionibacterium freudenreichii subsp. sherman

International audienceA gene potentially coding for exopolysaccharide (EPS) biosynthesis was identified in the genome sequence of Propionibacterium freudenreichii subsp. shermanii strain CIP 103027. The predicted protein product of this gene, designated Pps, best aligned with Tts, a polysaccharide synthase gene of Streptococcus pneumoniae type 37, which is responsible for the production of a glucan capsular polysaccharide. Pps also showed similarity with the putative glucan synthase Dps of Pediococcus damnosus and Oenococcus oeni. Specific primers were designed in order to detect the presence of pps in other P. freudenreichii subsp. shermanii strains by PCR amplification using total genomic DNA. The gene was detected in all the 12 strains tested. The same primers failed to detect the gene in an EPS-producing strain of Propionibacterium acidipropionicii, suggesting that the gene is not present or is sufficiently divergent in sequence to prevent amplification. Pps gene was totally sequenced for 8 strains, and seems to be very conserved, as less than 1 % of nucleotides differed from one strain to another. In order to evaluate pps gene expression, total RNA of 3 strains was extracted at different growth phases in YEL broth. The presence of mRNA corresponding to the gene was detected by RT-PCR in all 3 strains during exponential and stationary growth phases. This work is the first identification of a polysaccharide synthase gene of P. freudenreichii, and further studies need to be undertaken to measure the variability among strains in the production of capsular EPS

Lien entre lyse et lysogénie chez lactobacillus helveticus (mécanisme et impact sur l'affinage de l'emmental)

RENNES-Agrocampus-CRD (352382323) / SudocSudocFranceF

New probiotics: Dairy propionibacteria and the modulation of gut microbiota and physiology

This book underlines the importance of reciprocal interactions between probiotics and humans in terms of stress induction, epigenetic control of cellular responses, oxidative status, bio-active molecules biosynthesis, moonlighting proteins secretion, endogenous toxins neutralization, and several other biological functions. It explores how these responses can affect metabolism and metabolic-related disorders, gutbrain axis balance, mood, inflammatory, allergic and anti-infective reactions, cancer, and ageing. The book explores how probiotics create a dynamic and "fluid" network of signals able to control the balance between healthy and altered human status

New insights into physiology and metabolism of Propionibacterium freudenreichii

Dairy propionibacteria are Actinobacteria, mainly isolated from dairy environments. Propionibacterium freudenreichii has been used for a long time as a ripening culture in Swiss-type cheese manufacture, and is more and more considered for its potent probiotic effects. This review summarises the knowledge on the main P. freudenreichii pathways and the main features explaining its hardiness, and focuses on recent advances concerning its applications as a cheese ripening agent and as a probiotic for human health. Propionibacteria have a peculiar metabolism, characterised by the formation of propionic acid as main fermentation end-product. They have few nutritional requirements and are able to use a variety of carbon substrates. From the sequence of P. freudenreichii CIRM-BIA1T genome, many pathways were reconstituted, including the Wood–Werkman cycle, enzymes of the respiratory chain, synthesis pathways for all amino acids and many vitamins including vitamin B12. P. freudenreichii displays features allowing its long-term survival. It accumulates inorganic polyphosphate (polyP) as energy reserve, carbon storage compounds (glycogen), and compatible solutes such as trehalose. In cheese, P. freudenreichii plays an essential role in the production of a variety of flavour compounds, including not only propionic acid, but also free fatty acids released via lipolysis of milk glycerides and methyl-butanoic acids resulting from amino acid degradation. P. freudenreichii can exert health-promoting activities, such as a bifidogenic effect in the human gut and promising immunomodulatory effects. Many P. freudenreichii properties involved in adaptation, cheese ripening, bio-preservation and probiotic effects are highly strain-dependent. The elucidation of the molecular mechanisms involved is now facilitated by the availability of genome sequence and molecular tools. It will help in the selection of the most appropriate strain for each application

Propionibacterium ssp.

 Mention d'édition : 2ème éditionPropionibacterium ssp

Propriétés anti-inflammatoires d'une protéine de surface de propionibacterium freudenreichii

The present invention relates to a polypeptide comprising or consisting of the amino acid sequence SEQ id n DEG :1, for use in the treatment or prevention of an inflammatory disease. The invention further encompasses a nucleic acid sequence encoding a polypeptide of the invention, a vector comprising a nucleic acid of the invention and a host cell comprising a nucleic acid sequence and/or a vector of the invention, for use in the treatment or prevention of an inflammatory disease. The invention also concerns a pharmaceutical composition or a food composition comprising a polypeptide, a nucleic acid sequence, a vector or a host cell of the invention and a pharmaceutically acceptable carrier or a dairy product, for the treatment of inflammatory disease. The invention also relates a method for the screening of bacteria having immunomodulatory properties.La présente invention concerne un polypeptide comprenant ou se composant de la séquence d'acides aminés SEQ id n°:1, à utiliser dans le traitement ou la prévention d'une maladie inflammatoire. L'invention concerne, en outre, une séquence d'acides nucléiques codant un polypeptide de l'invention, un vecteur comprenant un acide nucléique de l'invention et une cellule hôte comprenant une séquence d'acides nucléiques et/ou un vecteur de l'invention, à utiliser dans le traitement ou la prévention d'une maladie inflammatoire. L'invention concerne aussi une composition pharmaceutique ou une composition alimentaire comprenant un polypeptide, une séquence d'acides nucléiques, un vecteur ou une cellule hôte de l'invention et un support pharmaceutiquement acceptable ou un produit laitier, pour le traitement d'une maladie inflammatoire. L'invention concerne également un procédé de criblage de bactéries ayant des propriétés immunomodulatrices

Propionibacterium ssp.

 Mention d'édition : 2ème éditionPropionibacterium ssp

Surface properties associated with the production of polysaccharides in the food bacteria Propionibacterium freudenreichii

International audienceThis study explores the production of polysaccharides (PS) in the strain Pf2289 of the food species Propioni-bacterium freudenreichii. Pf2289 presents characteristics atypical of the species: a molar-shaped morphotype upon plating, and cells strongly aggregative in liquid medium. When plating Pf2289, another morphotype was observed with a 4% frequency of appearance: round-shaped colonies, typical of the species. A clone was isolated, designated Pf456. No reversibility of Pf456 towards the molar-shaped morphotype was observed. Pf2289 was shown to produce a surface polysaccharide (PS) bound to the cell wall, mainly during the stationary growth phase. Meanwhile, Pf456 had lost the ability to produce the PS. AFM images of Pf2289 showed that entangled filaments spread over the whole surface of the bacteria, whereas Pf456 exhibited a smooth surface. Adhesion force maps, performed with concanavalin-A grafted probes, revealed twice as much adhesion of Pf2289 to concanavalin-A compared to Pf456. Furthermore, the length of PS molecules surrounding Pf2289 measured at least 7 μm, whereas it only reached 1 μm in Pf456. Finally, the presence of PS had a strong impact on adhesion properties: Pf2289 did not adhere to hydrophobic surfaces, whereas Pf456 showed strong adhesion