A novel acetone-degrading, nitrate-reducing bacterium, strain KN Bun08, was isolated from an enrichment culture with butanone and nitrate as the sole sources of carbon and energy. The cells were motile short rods, 0.5 to 1 by 1 to 2 m in size, which gave Gram-positive staining results in the exponential growth phase and Gram-negative staining results in the stationary-growth phase. Based on 16S rRNA gene sequence analysis, the isolate was assigned to the genus Alicycliphilus. Besides butanone and acetone, the strain used numerous fatty acids as substrates. An ATP-dependent acetone-carboxylating enzyme was enriched from cell extracts of this bacterium and of Alicycliphilus denitrificans K601 T by two subsequent DEAE Sepharose column procedures. For comparison, acetone carboxylases were enriched from two additional nitrate-reducing bacterial species, Paracoccus denitrificans and P. pantotrophus. The products of the carboxylase reaction were acetoacetate and AMP rather than ADP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of cell extracts and of the various enzyme preparations revealed bands corresponding to molecular masses of 85, 78, and 20 kDa, suggesting similarities to the acetone carboxylase enzymes described in detail for the aerobic bacterium Xanthobacter autotrophicus strain Py2 (85.3, 78.3, and 19.6 kDa) and the phototrophic bacterium Rhodobacter capsulatus. Protein bands were excised and compared by mass spectrometry with those of acetone carboxylases of aerobic bacteria. The results document the finding that the nitrate-reducing bacteria studied here use acetone-carboxylating enzymes similar to those of aerobic and phototrophic bacteria.Aerobic degradation of acetone and other methyl ketones was observed first with hydrocarbon-utilizing bacteria (13). Acetone is degraded by some bacteria (28) and mammalian liver cells via oxygenase-dependent hydroxylation to acetol (for a review, see reference 11). Anaerobic enrichments with acetone in the presence of sulfate resulted in selection of Desulfococcus multivorans-like cells (29), and Desulfococcus biautus was isolated using acetone plus sulfate (22). Nitrate-dependent acetone oxidation was documented with Paracoccus pantotrophus (3) and an unidentified bacterium, strain BunN (20). Also, the anoxygenic phototrophic bacterium Rhodobacter capsulatus was previously shown to grow with acetone as its sole electron source (2).Acetone activation by an initial oxygen-dependent hydroxylation to acetol was documented with hydrocarbon-utilizing bacteria (13). Anaerobic acetone degraders carboxylate acetone to an acetoacetyl residue, and this type of reaction was documented first with a methanogenic enrichment culture (19). Dependence of acetone utilization on the availability of CO 2 was also shown with nitrate-reducing and sulfate-reducing bacteria, with Paracoccus pantotrophus (3), and with Rhodobacter capsulatus and other phototrophs (2).The reaction mechanism of acetone carboxylation was studied into detail with the aerobic Xanthob...
In this study, we isolated Lactobacillus spp. from bovine raw milk and artisanal cheese from southern Brazil, and evaluated their technological and probiotic potential to select new isolates for producing healthy fermented dairy foods with differentiated tastes and flavours. We obtained 48 new lactobacilli isolates, which were isolated from raw milk (38) and cheese (10). These bacterial isolates were closely related with ten species: Lactobacillus paracasei (50% of the isolates), L. parabuchneri (15%), L. pentosus (13%), L. zeae (4%), L. plantarum (4%), L. otakiensis (4%), L. casei (4%), L. harbinensis (2%), L. diolivorans (2%), and L. rhamnosus (2%). Isolates CH112 and CH131 showed the greatest acidification potential, reducing the pH of milk to below 5.3 after incubation for 6 h at 32 °C. Considering proteolytic activity and diacetyl production, isolates ML88a, ML04, and ML12 showed the most promising results. Isolate ML12 showed 100% survival rate when inoculated in gastric juice at pH 2.5. The evaluation of antibacterial activity of the lactobacilli showed that the pathogens Listeria monocytogenes, Staphylococcus aureus, Salmonella enteritidis, and Salmonella Typhimurium were strongly inhibited by the pure lactobacilli cultures. Five Lactobacillus isolates (ML01, ML04, ML12, ML88, and CH139) showed both technological and probiotic characteristics. Principal Component Analysis (PCA) was used to investigate correlations among technological and probiotic characteristics, and identified new promising lactobacilli isolates for exploring their characteristics. This study reveals the importance of selecting new microorganisms with potential applicability in the food industry for developing functional foods with differentiated aromas and flavours. Keyword Brazil. Fermented dairy foods. Functional foods. Lactic acid bacteria. Lactobacilli isolates. Starter culture.
Selenium is an essential micronutrient for living beings, as it helps to maintain the normal physiological functions of the organism. The numerous discoveries involving the importance of this element to the health of human beings have fostered interest in research to develop enriched and functional foods. The present study evaluated the potential for bacterial strains of Enterococcus faecalis (CH121 and CH124), Lactobacillus parabuchneri (ML4), Lactobacillus paracasei (ML13, ML33, CH135, and CH139), and Lactobacillus plantarum (CH131) to bioaccumulate Se in their biomass by adding different concentrations of sodium selenite (30 to 200 mg/L) to the culture medium. Quantification of Se with UV and visible molecular absorption spectroscopy showed that the investigated bacteria were able to bioaccumulate this micromineral into their biomass. Two of the L. paracasei strains (ML13 and CH135) bioaccumulated the highest Se concentrations (38.1 ± 1.7 mg/g and 40.7 ± 1.1 mg/g, respectively) after culture in the presence of 150 mg/L of Se. This bioaccumulation potential has applications in the development of dairy products and may be an alternative Se source in the diets of humans and other animals.
The fourth author's last name was incorrect; it should be Cassiano Ricardo Brandt.
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