Graciela Rollán

Effect of galactose and glucose on the exopolysaccharide production and the activities of biosynthetic enzymes in Lactobacillus casei CRL 87.

Aims: The objective of this work was to study the influence of the sugar source on exopolysaccharide (EPS) production and the activities of the enzymes involved in the synthesis of sugar nucleotides in Lactobacillus casei CRL 87. The relationship between these enzymes and EPS formation was determined.

Inhibition of Citrus Fungal Pathogens by Using Lactic Acid Bacteria

The effect of lactic acid bacteria (LAB) on pathogenic fungi was evaluated and the metabolites involved in the antifungal effect were characterized. Penicillium digitatum (INTA 1 to INTA 7) and Geotrichum citri-aurantii (INTA 8) isolated from decayed lemon from commercial packinghouses were treated with imazalil and guazatine to obtain strains resistant to these fungicides. The most resistant strains (4 fungal strains) were selected for evaluating the antifungal activity of 33 LAB strains, among which only 8 strains gave positive results. The antifungal activity of these LAB strains was related to the production of lactic acid, acetic acid, and phenyllactic acid (PLA). A central composite design and the response surface methodology were used to evaluate the inhibitory effect of the organic acids produced by the LAB cultures. The antifungal activity of lactic acid was directly related to its concentration; however, acetic acid and PLA showed a peak of activity at 52.5 and 0.8 mM, respectively, with inhibition rates similar to those obtained with Serenade((R)) (3.0 ppm) imazalil (50 ppm) and guazatine (50 ppm). Beyond the peak of activity, a reduction in effectiveness of both acetic acid and PLA was observed. Comparing the inhibition rate of the organic acids, PLA was about 66- and 600-fold more effective than acetic acid and lactic acid, respectively. This study presents evidences on the antifungal effect of selected LAB strains and their end products. Studies are currently being undertaken to evaluate the effectiveness in preventing postharvest diseases on citrus fruits.

Proteolytic activity and reduction of gliadin-like fractions by sourdough lactobacilli

Aims:  To characterize the peptide hydrolase system of Lactobacillus plantarum CRL 759 and CRL 778 and evaluate their proteolytic activity in reducing gliadin‐like fractions.

A combination of two lactic acid bacteria improves the hydrolysis of gliadin during wheat dough fermentation

The evaluation of gliadin hydrolysis during dough fermentation by using two lactic acid bacteria, Lactobacillus plantarum CRL 775 and Pediococcus pentosaceus CRL 792, as pooled cell suspension (LAB) or cell free extract (CFE) was undertaken. The CFE pool produced a greater (121%) increase in amino acid concentration than the LAB pool (70-80%). These results were correlated with the decrease (76,100 and 64,300 ppm) in the gliadin concentration of doughs supplemented with CFE and LAB, respectively, compared to control doughs. The use of LAB peptidases seemed to be a viable technologic alternative to reduce the gliadin concentration in wheat dough without using living bacteria as starter.

Biodiversity and technological-functional potential of lactic acid bacteria isolated from spontaneously fermented quinoa sourdoughs

To analyse lactic acid bacteria (LAB) diversity and technological‐functional and safety properties of strains present during spontaneous fermented quinoa sourdoughs.

Lactobacillus reuteri CRL 1100 as starter culture for wheat dough fermentation

The effect of sucrose on the fermentation balance of Lactobacillus reuteri CRL 1100 and the invertase activity of this strain in wheat dough and culture medium (MRSs) was evaluated. The enzyme activity was dependent on the environmental pH releasing glucose and fructose from sucrose hydrolysis. Glucose was used as carbon source, while fructose was mainly used as electron acceptor to produce mannitol up to 10h of fermentation. Thereafter, fructose seemed to be metabolized by the heterofermentative pathway, which determined an increase in the concentration of acetate (6 mmol l(-1)), lactate (2 mmol l(-1)) and ethanol (1 mmol l(-1)) and the lack of mannitol formation after glucose depletion. The fermentation balance of Lb. reuteri CRL 1100 during the dough fermentation resulted in lower (63%) ethanol, higher (75%) acetate production and soluble carbohydrates concentrations, like MRSs cultures. This fermentation profile would be important to obtain an optimal growth of yeast and the optimal bread flavor and taste.

The peptide hydrolase system of Lactobacillus reuteri

Peptide hydrolase system of Lactobacillus reuteri CRL 1098, a lactic acid bacteria of sourdough origin, was investigated. This microorganism has a broad range of peptidases consisting of an active aminopeptidase, X-Prolyl-dipeptidylaminopeptidase, dipeptidase and tripeptidase. Aminopeptidase, iminopeptidase and endopeptidase are most likely located in the cytoplasmic fraction showing no detectable association with the cell membrane, while dipeptidase and tripeptidase are mainly associated with the latter fraction. The peptidases are metalloenzymes activated by Co2+ and inhibited by Cu2+, Hg2+, Cd2+ and by metal-complexing reagents. The aminopeptidase activity inhibited by EDTA can be restored by Mn2+ while that of di- and tripeptidase treated with 1,10-phenantroline can be restored by Zn2+ and Co2+, respectively.

Biodiversity and technological potential of lactic acid bacteria isolated from spontaneously fermented amaranth sourdough.

Spontaneous fermented sourdoughs prepared from amaranth flour were investigated for the presence of autochthonous lactic acid bacteria (LAB) predominating microbiota. The doughs were fermented with daily backslopping on a laboratory scale at 30°C for 10 days. LAB counts ranged from 2·60 to 8·54 log CFU g−1 with a pH declined from 6·2 to 3·8 throughout fermentation. The combined use of randomly amplified polymorphic DNA (RAPD)‐PCR analysis and sequence analysis of 16S rRNA was applied for LAB intraspecies differentiation and taxonomic identification, respectively. Enterococcus, Pediococcus and Lactobacillus species were present in amaranth sourdoughs (AS). After the first refreshment step, Lactobacillus plantarum dominated AS until the end of fermentation. In coincidence, when DGGE analysis was performed, the occurrence of a progressive change in bacterial communities allowed the selection of Lact. plantarum as a dominant species. Moreover, technological, functional and safety characteristics of representative RAPD‐biotypes were investigated. Lact. plantarum CRL1898 was selected as a potential candidate for gluten‐free amaranth sourdough starter.

Ancestral Andean grain quinoa as source of lactic acid bacteria capable to degrade phytate and produce B-group vitamins

The lactic acid bacteria (LAB) microbiota of quinoa grains (QG) and spontaneous sourdough (QSS) was evaluated. Different strains of Lactobacillus (L.) plantarum (7), L. rhamnosus (5), L. sakei (1), Pediococcus (Ped.) pentosaceus (9), Leuconostoc (Leuc.) mesenteroides (1), Enterococcus (E.) casseliflavus (2), E. mundtii (3), E. hirae (1), E. gallinarum (12), Enterococcus sp. (1), and E. hermanniensis (2) were isolated, identified and characterized. Only four strains isolated from QSS and eight strains isolated from QG showed amylolytic activity. L. plantarum CRL 1973 and CRL 1970, L. rhamnosus CRL 1972 and L. sakei CRL 1978 produced elevated concentrations of folate with strain CRL 1973 producing the highest concentration (143±6ng/ml). L. rhamnosus, isolated from QSS, was the LAB species that produced the most elevated concentrations of total riboflavin (>270ng/ml) with strain CRL 1963 producing the highest amounts (360±10ng/ml). Phytase activity, evaluated in forty-four LAB strains from quinoa, was predominantly detected in L. rhamnosus and Enterococci strains with the highest activities observed in E. mundtii CRL 2007 (957±25U/ml) followed by E. casseliflavus CRL 1988 (684±38U/ml), Leuc. mesenteroides CRL 2012 (617±38U/ml) and L. rhamnosus CRL 1983 (606±79U/ml). In conclusion, this study shows that a diverse LAB microbiota is present in quinoa with important properties; these microorganisms could be used as potential starter cultures to increase the nutritional and functional properties of Andean grains based foods.

Microbial applications in the biopreservation of cereal products

Abstract: Microorganisms play a central role in the manufacture of nutritious fermented foods and beverages. They are also of importance in the maintenance of food storage, food safety, and quality. Cereals and cereal-based products are usually spoiled by bacteria and moulds resulting either in unpleasant texture, smell and odour, or in toxic or poisoning products. The food preservation mediated by microorganisms and fermentation technologies has been attributed to the production of metabolites such as organic acids, hydrogen peroxide, alcohols and antibiotics. Antibiosis by bacteriocins offers potential applications in food biopreservation, while the use of enzymes derived from phages (i.e. enzybiotics) remains as a promising option.