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1.
Front Microbiol ; 9: 2256, 2018.
Article in English | MEDLINE | ID: mdl-30319575

ABSTRACT

Vitamin B12 or cobalamin is an essential metabolite for humans, which makes it an interesting compound for many research groups that focus in different producer-strains synthesis pathways. In this work, we report the influence of key intermediaries for cobalamin synthesis added to the culture medium in two Lactobacillus (L.) strains, L. reuteri CRL 1098 and L. coryniformis CRL 1001. Here, we report that addition of Co2+ and 5,6-dimethylbenzimidazole increased the corrinoid compounds production in both strains while addition of L-threonine increased only the corrinoid compounds production by CRL 1001 strain. Then, we purified and characterized by LC-MS the corrinoid compounds obtained. Physiological studies besides in silico analysis revealed that L. reuteri CRL 1098 and L. coryniformis CRL 1001 follow different pathways for the last steps of the corrinoid compounds synthesis.

2.
J Basic Microbiol ; 58(4): 296-301, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29430723

ABSTRACT

In artisanal tanneries, the skins are immersed in cereals fermented by natural microbial flora in order to reduce the pH of the skin, an essential condition for carrying out the final step. The environmental thermal variation alters the fermentation process and affects the quality of the final product. The aim of this work was to isolate lactic acid bacteria from cereals mixture fermented in an artisanal tannery and to evaluate in vitro the acidifying activity of the strains as a first step for the formulation of a starter culture. In most samples, a prevalence of cocci (95%) was observed with respect to bacilli. The best acidifying strains were identified by phenotypic and genotypic techniques as Enterococcus faecium CRL 1943 (rapid acidification at 37 °C) and Leuconostoc citreum CRL 1945 (high acidifying activity at 18 °C). In addition, the biomass production of the selected strains was analyzed at free and controlled pH (bioreactors 1.5 L). The production of biomass was optimal at controlled pH, with a higher growth (0.5-1.1 log units). Both strains were compatible, allowing their inclusion in a mixed culture. These lactic strains could contribute to the systematization of the tanning process.


Subject(s)
Lactobacillales/isolation & purification , Lactobacillales/metabolism , Tanning/methods , Bioreactors , DNA, Bacterial/genetics , Edible Grain/microbiology , Enterococcus faecium/genetics , Enterococcus faecium/growth & development , Enterococcus faecium/isolation & purification , Enterococcus faecium/metabolism , Fermentation , Genotype , Hydrogen-Ion Concentration , Lactic Acid/biosynthesis , Lactobacillales/genetics , Lactobacillales/growth & development , Leuconostoc/genetics , Leuconostoc/growth & development , Leuconostoc/isolation & purification , Leuconostoc/metabolism , Phenotype , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Temperature
3.
BMC Microbiol ; 16(1): 240, 2016 Oct 13.
Article in English | MEDLINE | ID: mdl-27737643

ABSTRACT

BACKGROUND: Despite the fact that most vitamins are present in a variety of foods, malnutrition, unbalanced diets or insufficient intake of foods are still the cause of vitamin deficiencies in humans in some countries. Vitamin B12 (Cobalamin) is a complex compound that is only naturally produced by bacteria and archea. It has been reported that certain strains belonging to lactic acid bacteria group are capable of synthesized water-soluble vitamins such as those included in the B-group, as vitamin B12. In this context, the goal of the present paper was to evaluate and characterize the production of vitamin B12 in Lactobacillus coryniformis CRL 1001, a heterofermentative strain isolated from silage. RESULTS: Cell extract of L. coryniformis CRL 1001, isolated from silage, is able to correct the coenzyme B12 requirement of Salmonella enterica serovar Typhimurium AR 2680 in minimal medium. The chemical characterization of the corrinoid-like molecule isolated from CRL 1001 cell extract using HPLC and mass spectrometry is reported. The majority of the corrinoid produced by this strain has adenine like Coα-ligand instead 5,6-dimethylbenzimidazole. Genomic studies revealed the presence of the complete machinery of the anaerobic biosynthesis pathway of coenzyme B12. The detected genes encode all proteins for the corrin ring biosynthesis and for the binding of upper (ß) and lower (α) ligands in one continuous stretch of the chromosome. CONCLUSIONS: The results here described show for the first time that L. coryniformis subsp. coryniformis CRL 1001 is able to produce pseudocobalamin containing adenine instead of 5,6-dimethlbenzimidazole in the Coα-ligand. Genomic analysis allowed the identification and characterization of the complete de novo biosynthetic pathway of the corrinoid produced by the CRL 1001 strain.


Subject(s)
Lactobacillus/genetics , Lactobacillus/metabolism , Multigene Family , Vitamin B 12/biosynthesis , Vitamin B 12/genetics , Adenine/metabolism , Anaerobiosis , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Base Sequence , Benzimidazoles/metabolism , Chromatography, High Pressure Liquid/methods , Cobamides/genetics , Cobamides/metabolism , Genes, Bacterial , Lactobacillus/isolation & purification , Mass Spectrometry/methods , Open Reading Frames , Salmonella enterica/genetics , Salmonella enterica/metabolism , Salmonella typhimurium/genetics , Sequence Analysis, Protein , Silage/microbiology , Vitamin B 12/chemistry , Vitamin B 12/isolation & purification
4.
Genome Announc ; 4(4)2016 Aug 25.
Article in English | MEDLINE | ID: mdl-27563038

ABSTRACT

We report here the draft genome sequence of Lactobacillus reuteri strain CRL 1098. This strain represents an interesting candidate for functional food development because of its proven probiotic properties. The draft genome sequence is composed of 1,969,471 bp assembled into 45 contigs and an average G+C content of 38.8%.

5.
J Sci Food Agric ; 91(13): 2355-62, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21604277

ABSTRACT

BACKGROUND: Lactobacillus rhamnosus CRL1505, a strain of goats' milk origin, is able to stimulate mucosal immunity and protect immunocompetent mice from intestinal and respiratory infections. RESULTS: In this work we developed and characterized a fermented goats' milk containing L. rhamnosus CRL1505, and we demonstrated in a model of immunosuppression in mice that the final dairy product preserves the immunomodulatory properties of the strain. L. rhamnosus CRL1505 survived the manufacturing process of fermented milk and maintained a viability of 10(6) cfu g(-1) during storage. The fermented goats' milk was accepted by 90.48% of the panelists and was considered as having an acid taste and pleasant aroma. We also demonstrated that the developed product, used as a supplement during the repletion of immunocompromised malnourished mice, was effective in accelerating the recovery of clinical parameters altered by malnutrition and to induce increased resistance against intestinal and respiratory infections. CONCLUSION: Goats' milk fermented with L. rhamnosus CRL1505 could be manufactured as an alternative probiotic dairy product since this new food has the ability to stimulate the common mucosal immune system and to improve defenses against respiratory and intestinal infections.


Subject(s)
Cultured Milk Products/microbiology , Immunity, Mucosal , Immunocompromised Host , Immunomodulation , Lacticaseibacillus rhamnosus/immunology , Adolescent , Animals , Argentina , Child , Disease Resistance , Food Handling , Food Preferences , Goats , Humans , Lacticaseibacillus rhamnosus/growth & development , Lacticaseibacillus rhamnosus/isolation & purification , Lacticaseibacillus rhamnosus/metabolism , Male , Malnutrition/diet therapy , Malnutrition/immunology , Malnutrition/physiopathology , Mice , Microbial Viability , Pneumonia, Pneumococcal/immunology , Pneumonia, Pneumococcal/prevention & control , Salmonella Food Poisoning/immunology , Salmonella Food Poisoning/prevention & control
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