Differential flora in the microenvironment of lung tumor and paired adjacent normal tissues.

Recent evidence demonstrates the existence of diversified microbiota in the lung. However, the effect of lung carcinogenesis on the flora in lung microenvironment has yet not been well investigated. In this study, we surveyed the microbial composition and diversity in lung tumor and paired adjacent normal tissues obtained from 55 lung cancer patients to test whether any specific tumor-associated microbial features in lung microenvironment can be identified. Compared with non-malignant adjacent tissues, the tumor samples showed significantly lower community richness (α diversity), but no significant difference in overall microbiome dissimilarity (ß diversity). Strong intrasubject correlations were observed between tumor sample and its paired non-malignant adjacent tissues. In addition, correlation network analysis found more significant taxa-taxa correlations (adjusted q-value < 0.05) in tumor microenvironment than non-malignant adjacent tissues. At taxa level, we found Propionibacterium genus were significantly reduced in tumor tissues compared with non-malignant adjacent tissues. In summary, the microbiota in tumor tissues showed the lower richness, higher taxa-taxa interaction, and reduction of potential pro-inflammatory microbial genera compared with non-malignant tissues, suggesting the potential link between the tumor microbiota and the altered tumor microenvironment for the further investigation.

Comparative genomics reveals distinct host-interacting traits of three major human-associated propionibacteria.

BACKGROUND: Propionibacteria are part of the human microbiota. Many studies have addressed the predominant colonizer of sebaceous follicles of the skin, Propionibacterium acnes, and investigated its association with the skin disorder acne vulgaris, and lately with prostate cancer. Much less is known about two other propionibacterial species frequently found on human tissue sites, Propionibacterium granulosum and Propionibacterium avidum. Here we analyzed two and three genomes of P. granulosum and P. avidum, respectively, and compared them to two genomes of P. acnes; we further highlight differences among the three cutaneous species with proteomic and microscopy approaches. RESULTS: Electron and atomic force microscopy revealed an exopolysaccharide (EPS)-like structure surrounding P. avidum cells, that is absent in P. acnes and P. granulosum. In contrast, P. granulosum possesses pili-like appendices, which was confirmed by surface proteome analysis. The corresponding genes were identified; they are clustered with genes encoding sortases. Both, P. granulosum and P. avidum lack surface or secreted proteins for predicted host-interacting factors of P. acnes, including several CAMP factors, sialidases, dermatan-sulphate adhesins, hyaluronidase and a SH3 domain-containing lipoprotein; accordingly, only P. acnes exhibits neuraminidase and hyaluronidase activities. These functions are encoded on previously unrecognized island-like regions in the genome of P. acnes. CONCLUSIONS: Despite their omnipresence on human skin little is known about the role of cutaneous propionibacteria. All three species are associated with a variety of diseases, including postoperative and device-related abscesses and infections. We showed that the three organisms have evolved distinct features to interact with their human host. Whereas P. avidum and P. granulosum produce an EPS-like surface structure and pili-like appendices, respectively, P. acnes possesses a number of unique surface-exposed proteins with host-interacting properties. The different surface properties of the three cutaneous propionibacteria are likely to determine their colonizing ability and pathogenic potential on the skin and at non-skin sites.

Evaluation of positive interaction for cell growth between Bifidobacterium adolescentis and Propionibacterium freudenreichii using a co-cultivation system with two microfiltration modules.

Using a co-cultivation system developed previously, positive interaction for cell growth between Bifidobacterium adolescentis and Propionibacterium freudenreichii was evaluated. The total dry cell weight (DCW) of these two strains obtained in the co-cultivation system was 1.5-1.7-fold of the sum of the DCWs obtained in two single cultivations of each bacterium.

An economical biorefinery process for propionic acid production from glycerol and potato juice using high cell density fermentation.

An economically sustainable process was developed for propionic acid production by fermentation of glycerol using Propionibacterium acidipropionici and potato juice, a by-product of starch processing, as a nitrogen/vitamin source. The fermentation was done as high-cell-density sequential batches with cell recycle. Propionic acid production and glycerol consumption rates were dependent on initial biomass concentration, and reached a maximum of 1.42 and 2.30 g L(-1) h(-1), respectively, from 50 g L(-1) glycerol at initial cell density of 23.7 gCDW L(-1). Halving the concentration of nitrogen/vitamin source resulted in reduction of acetic and succinic acids yields by ~39% each. At glycerol concentrations of 85 and 120 g L(-1), respectively, 43.8 and 50.8 g L(-1) propionic acid were obtained at a rate of 0.88 and 0.29 g L(-1) h(-1) and yield of 84 and 78 mol%. Succinic acid was 13 g% of propionic acid and could represent a potential co-product covering the cost of nitrogen/vitamin source.

The first dairy product exclusively fermented by Propionibacterium freudenreichii: a new vector to study probiotic potentialities in vivo.

Dairy propionibacteria display probiotic properties which require high populations of live and metabolically active propionibacteria in the colon. In this context, the probiotic vector determines probiotic efficiency. Fermented dairy products protect propionibacteria against digestive stresses and generally contain a complex mixture of lactic and propionic acid bacteria. This does not allow the identification of dairy propionibacteria specific beneficial effects. The aim of this study was to develop a dairy product exclusively fermented by dairy propionibacteria. As they grow poorly in milk, we determined their nutritional requirements concerning carbon and nitrogen by supplementing milk ultrafiltrate (UF) with different concentrations of lactate and casein hydrolysate. Milk or UF supplemented with 50 mM lactate and 5 g L(-1) casein hydrolysate allowed growth of all dairy propionibacteria studied. In these new fermented dairy products, dairy propionibacteria remained viable and stress-tolerant in vitro during minimum 15 days at 4 °C. The efficiency of milk fermented by the most tolerant Propionibacterium freudenreichii strain was evaluated in piglets. Viability and SCFA content in the colon evidenced survival and metabolic activity of P. freudenreichii. This work results in the design of a new food grade vector, which will allow preclinical and clinical trials.

Batch- and continuous propionic acid production from glycerol using free and immobilized cells of Propionibacterium acidipropionici.

Propionic acid production from glycerol was studied using Propionibacterium acidipropionici DSM 4900 cells immobilized on polyethylenimine-treated Poraver (PEI-Poraver) and Luffa (PEI-Luffa), respectively. Using PEI-Luffa, the average productivity, yield and concentration of propionic acid from 40 g L(-1) glycerol were 0.29 g L(-1) h(-1), 0.74 mol(PA) mol(Gly)(-1) and 20 g L(-1), respectively, after four consecutive recycle-batches. PEI-Poraver supported attachment of 31 times higher amounts of cells than PEI-Luffa and produced 20, 28 and 35 g L(-1) propionic acid from 40, 65 and 85 g L(-1) glycerol, respectively (0.61 mol(PA) mol(Gly)(-1)). The corresponding production rates were 0.86, 0.43 and 0.35 g L(-1) h(-1), which are the highest reported from glycerol via batch or fed-batch fermentations for equivalent propionic acid concentrations. Using a continuous mode of operation at a dilution rate of 0.1 h(-1), cell washout was observed in the bioreactor with free cells; however, propionic acid productivity, yield and concentration were 1.40 g L(-1) h(-1), 0.86 mol(PA) mol(Gly)(-1), and 15 g L(-1), respectively, using immobilized cells in the PEI-Poraver bioreactor. The choice of the immobilization matrix can thus significantly influence the fermentation efficiency and profile. The bioreactor using cells immobilized on PEI-Poraver allowed the fermentation of higher glycerol concentrations and provided stable and higher fermentation rates than that using free cells or the cells immobilized on PEI-Luffa.

Improving the productivity of propionic acid with FBB-immobilized cells of an adapted acid-tolerant Propionibacterium acidipropionici.

Propionic acid is an important short-chain fatty acid with many applications, but its large-scale bioproduction was hindered by the low productivity. An adapted acid-tolerant Propionibacterium acidipropionici CGMCC 1.2230 strain was selected to produce propionic acid with a relatively high productivity (0.29 g/(Lh)) in the free-cell fermentation. Further immobilized-cell fermentation in fibrous-bed bioreactor (FBB) supported high-level repeated batch fermentations with a high productivity of 0.96 g/(Lh). The FBB also presents the potential to increase final propionic acid concentration by using glucose feeding strategy. The propionic acid concentration was increased to 51.2g/L in the fed-batch fermentation with the productivity of 0.71 g/(Lh). By adopting the above strategies, sugarcane bagasse hydrolysate could support the production of propionic acid with high productivity in the repeat-batch and fed-batch fermentations. The present work would pave one road to the accomplishment of large-scale bioproduction of propionic acid from renewable resources.

Microbial production of propionic acid with Propionibacterium freudenreichii using an anion exchanger-based in situ product recovery (ISPR) process with direct and indirect contact of cells.

The recovery of an inhibiting product from a bioreactor soon after its formation is an important issue in industrial bioprocess development. In the present study, the potential of the anion exchanger-based in situ product recovery (ISPR) technique for the biocatalytic production of propionic acid was discussed. The focus of the current work was the selection of a suitable configuration of metabolically active cells for application in propionic acid production. Accumulation of propionic acid in fermentation broth caused feedback inhibition of the growth and biotransformation activity of Propionibacterium freudenreichii CICC 10019. Relevant product inhibition kinetics was discussed, and the results showed that keeping the aqueous propionic acid concentration below 10.02 g L⁻¹ was an essential prerequisite for ISPR process. A batch study, in which three ISPR configuration mode designs were compared, was conducted. The comparison indicated that employing an external direct mode had significant advantages over other modes in terms of increased productivity and product yield, with a corresponding decrease in the number of downstream processing steps, as well as in substrate consumption. The fed-batch culture using an external direct mode for the continuous accumulation of propionic acid resulted in a cumulative propionic acid concentration of 62.5 g L⁻¹, with a corresponding product yield of 0.78 g propionic acid/g glucose.

Engineering Propionibacterium acidipropionici for enhanced propionic acid tolerance and fermentation.

Propionibacterium acidipropionici, a Gram-positive, anaerobic bacterium, has been the most used species for propionic acid production from sugars. In this study, the metabolically engineered mutant ACK-Tet, which has its acetate kinase gene knocked out from the chromosome, was immobilized and adapted in a fibrous bed bioreactor (FBB) to increase its acid tolerance and ability to produce propionic acid at a high final concentration in fed-batch fermentation. After about 3 months adaptation in the FBB, the propionic acid concentration in the fermentation broth reached approximately 100 g/L, which was much higher than the highest concentration of approximately 71 g/L previously attained with the wild-type in the FBB. To understand the mechanism and factors contributing to the enhanced acid tolerance, adapted mutant cells were harvested from the FBB and characterized for their morphology, growth inhibition by propionic acid, protein expression profiles as observed in SDS-PAGE, and H+-ATPase activity, which is related to the proton pumping and cell's ability to control its intracellular pH gradient. The adapted mutant obtained from the FBB showed significantly reduced growth sensitivity to propionic acid inhibition, increased H+-ATPase expression and activity, and significantly elongated rod morphology.

[A positive effect of Propionibacterium freudenreichii on the growth of Saccharomyces cerevisiae during their cocultivation].

The growth pattern of Saccharomyces cerevisiae and Propionibacterium freudenreichii ssp. shermanii (P. shermanii; propionic acid bacteria, PABs) during cocultivation in liquid media depended on the ratio of the cells in the inoculum. An increase in the growth rate of S. cerevisiae was observed at a PAB to yeast ratio of approximately 3 : 1; higher ratios exerted adverse effects on yeast growth. The culture liquid of 18- to 24-h (young) cultures of PABs stimulated yeast growth. Although yeast growth-stimulating exometabolites of PABs were not high-molecular-weight compounds, they were thermolabile. When present in the medium at concentrations of up to 1.5%, the antimicrobial agent sodium propionate did not interfere with S. cerevisiae growth; however, it completely inhibited the growth of B. subtilis at a concentration of 0.2%.

Behaviour of Propionibacterium shermanii in alkaline medium.

Physiological and morphological properties of Propionibacterium shermanii were estimated in a liquid semisynthetic medium of initial pH between 7-10. Prolongation of the lag phase and a shift of the stationary phase occurred at higher initial pH values of the medium. The growth response of the strain depended on the way of glucose addition (either sterilized in the medium or added aseptically after sterilization). At pH 8.5 and higher the cells begin to form slime and capsules. The strain exhibited growth activity even at initial pH values of the medium 11.5

[Protective effect of P40 fraction of C. granulosum against leukemia L1210 in mice]. / Effet protecteur de la fraction P40 isolée de C. granulosum dans la leucémie L1210 de la souris.

Administration of glutaraldehyde treated L1210 leukemia cells, either alone or coupled with tetanus toxoid by means of glutaraldehyde as well as L1210 cells inactivated by mitomycin, did not induce appreciable protection against a tumorigenic dose of L1210 cells. On the other hand, injection of P40 fraction of C. granulosum induced non-specific resistance to L1210 leukemia and increased the efficiency of specific immunization by either glutaraldehyde treated L1210 cells or cells coupled with tetanus toxoid. Injection of Freund's complete adjuvant resulted in increase of rate of mortality after challenge with L1210 cells.