Although most vitamins are present in a variety of foods, human vitamin deficiencies still occur in many countries, mainly because of malnutrition not only as a result of insufficient food intake but also because of unbalanced diets. Even though most lactic acid bacteria (LAB) are auxotrophic for several vitamins, it is now known that certain strains have the capability to synthesize water‐soluble vitamins such as those included in the B‐group (folates, riboflavin and vitamin B12 amongst others). This review article will show the current knowledge of vitamin biosynthesis by LAB and show how the proper selection of starter cultures and probiotic strains could be useful in preventing clinical and subclinical vitamin deficiencies. Here, several examples will be presented where vitamin‐producing LAB led to the elaboration of novel fermented foods with increased and bioavailable vitamins. In addition, the use of genetic engineering strategies to increase vitamin production or to create novel vitamin‐producing strains will also be discussed. This review will show that the use of vitamin‐producing LAB could be a cost‐effective alternative to current vitamin fortification programmes and be useful in the elaboration of novel vitamin‐enriched products.

The three-dimensional crystal structure of the PduO-type corrinoid adenosyltransferase from Lactobacillus reuteri

We found that Lactobacillus reuteri CRL1098, a lactic acid bacterium isolated from sourdough, is able to produce cobalamin. The sugar-glycerol cofermentation in vitamin B 12 -free medium showed that this strain was able to reduce glycerol through a well-known cobalamin-dependent reaction with the formation of 1,3-propanediol as a final product. The cell extract of L. reuteri corrected the coenzyme B 12 requirement of Lactobacillus delbrueckii subsp. lactis ATCC 7830 and allowed the growth of Salmonella enterica serovar Typhimurium (metE cbiB) and Escherichia coli (metE) in minimal medium. Preliminary genetic studies of cobalamin biosynthesis genes from L. reuteri allowed the identification of cob genes which encode the CobA, CbiJ, and CbiK enzymes involved in the cobalamin pathway. The cobamide produced by L. reuteri, isolated in its cyanide form by using reverse-phase high-pressure liquid chromatography, showed a UV-visible spectrum identical to that of standard cyanocobalamin (vitamin B 12 ).

Swiss Albino mice were fed a diet enriched with fat to produce hypercholesterolemia. The further administration of Lactobacillus reuteri CRL 1098 (10(4) cells/d) to hypercholesterolemic mice for 7 d decreased total cholesterol by 38%, producing serum cholesterol concentrations similar to that of the control group (67.4 mg/ml). This low dose of L. reuteri caused a 40% reduction in triglycerides and a 20% increase in the ratio of high density lipoprotein to low density lipoprotein without bacterial translocation of the native microflora into the spleen and liver. These data suggest that L. reuteri CRL 1098 is an effective hypocholesterolemic adjuvant at a low cell concentration for mice.

Aims: The objective of this study was to evaluate the effect of bile salts and cholesterol in the lipid profile of Lactobacillus reuteri CRL 1098 and to determine the relationship existing between these changes: the in vitro removal of cholesterol and the tolerance of the cells to acid and cold stress. Methods and Results: Lactobacillus reuteri CRL 1098 was grown in the following media: MRS (deMan Rogosa Sharpe; M C , control medium), M B (M C with bile salts), M CH (M C with sterile cholesterol) and M BCH (M C with bile salts and cholesterol). Fatty acids were determined by analytical gas-liquid chromatography, and phospholipids and glycolipids by colorimetric techniques. The cells from different culture media were subjected to cold and acid stress. The M B cultures displayed a decrease in phospholipids and a low ratio of saturated : unsaturated fatty acids. The presence of the unusual C18 : 0,10-OH and C18 : 0,10-oxo fatty acids was the prominent characteristic of the bile salts growing cells. The relative increase in glycolipids and the changes in the fatty acids profiles of the M B cells would be responsible for the cholesterol remotion. The changes induced by bile salts in the lipid profile did not improve the tolerance of L. reuteri CRL 1098 to freezing and acid stress. Conclusions: The changes in lipid profiles reported in this study would play a key role in the response of Lactobacilli to environmental stress. Significance and Impact of the Study: This work provides useful information about the effect of bile salts on the cell membrane of L. reuteri, a probiotic enterolactobacillus. The steady-state response of the cells subjected to bile stress seems to be the appropriate model for evaluating the bacterial behaviour in detergent-containing gastrointestinal tracts, where the bile salts stress would presumably be continuous.

Aims: The objective of this work was to evaluate the fermentation pattern of and the exopolysaccharide (EPS) production by Lactobacillus helveticus ATCC 15807 in milk batch cultures under controlled pH (4á5, 5á0 and 6á2). Methods and Results: EPS concentration was estimated by the phenol/sulphuric acid method and the chemical composition of puri®ed EPS by HPLC. Fermentation products and residual sugars were determined by HPLC and enzymatic methods. The micro-organism shifted from a homofermentative to a heterofermentative pattern, producing acetate (9á5 and 5á8 mmol l ±1 ) at pH 5á0 and 6á2, respectively, and acetate (7á1 mmol l ±1 ) plus succinate (1á2 mmol l ±1 ) at pH 4á5. At pH 5á0 and 6á2, acetate derived from citrate while at pH 4á5 it came from both citrate and pyruvate splitting. The EPS has a MW of 10 5 ±10 6 and contains phosphate (81% in average), rhamnose (traces), and glucose and galactose in a ratio of 1 : 1 (pH 6á2) and 2 : 1 (pH 4á5 and 5á0). The highest production (549 mg l ±1 ) corresponded to pH 5á0 and the lowest (49 mg l ±1 ) to pH 6á2. Conclusions: The heterofermentative pattern of Lact. helveticus ATCC 15807 was linked to alternative pyruvate pathways and/or citrate metabolism according to the environmental pH. The EPS production was improved under low environmental pH conditions. Signi®cance and Impact of the Study: This work provides relevant information of the effect of pH on the metabolism of citrate and EPS production by Lact. helveticus. It may contribute to improve technological aspects of ropy and citrate-utilizing lactic acid bacteria.

Administration of Lactobacillus reuteri CRL 1098 (10(4) cells/d) to mice for 7 d before inducing hypercholesterolemia (by feeding mice with a fat-enriched diet for the subsequent 7 d) was evaluated. At this low dose, L. reuteri was effective in preventing hypercholesterolemia in mice, producing a 17% increase in the ratio of high-density lipoprotein to low-density lipoprotein. Total cholesterol and triglycerides decreased by 22 and 33%, respectively, in the group that was not fed the lactobacilli. The hypocholesterolemic effect produced by L. reuteri CRL 1098 might be considered as indirect evidence of the permanency of the lactobacilli in the gut.

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