Study of volatile compounds profiles in milk matrices using Enterococcus faecalis EstA and Rhizomucor miehei lipase.

The use of esterase/lipase enzymes of different origins in food industry is a widely employed strategy to enhance the formation of characteristic aromatic compounds derived from fat and diversify flavour. In the present work, we studied EstA enzyme of Enterococcus faecalis and a high purity Rhizomucor miehei lipase (Palatase). EstA was obtained recombinantly in Escherichia coli BL21 (DE3), and optimum esterase activity was detected at pH 6.75 and 40 °C. We evaluated the effect of the enzymes on milk mixtures prepared with different fat contents (2.8 and 6%) and structure (native or homogenized) on volatile compounds profiles. The milk fat structure before and after the application of low homogenization was characterized by dynamic light dispersion and microscopy. Native milk fat mixtures presented particles of 4.6 µm and 184 nm and homogenized mixtures had particles of 1.4 µm and 258 nm; microscopy images were in concordance with these results. Fifteen volatile compounds were identified, including ketones, esters, alcohols, and acids. We showed the key role of milk fat levels and microstructure in the nature of the volatile compounds produced by the R. miehei enzyme. Both in native or homogenized states, the highest content of fat favored a higher production of acids whereas the lowest fat level favored a higher esters production along with a more balanced volatile profile. For EstA enzyme, results showed a limited action on fat, as biosynthesis of esters only increased with the highest fat level homogenized.

Influence of commercial starter culture on fermentation dynamics and quality characteristics of yogurts obtained with different formulations.

BACKGROUND: Four commercial starter cultures containing Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (S1, S2, S3 and S4; S3 also contained Limosilactobacillus fermentum) were compared for fermentation, volatile flavor compounds, physicochemical parameters and microbiology, in yogurt prepared from three milk base formulations with increased protein (B1, B2 and B3). RESULTS: The fermentation patterns differed among starters, with Yoflex Mild 1.0 (S4) and SLB95 (S2) showing the longest fermentation time, depending on the formulation. At 21 days, S. thermophilus counts were similar among starters and higher than 8.52 log CFU mL-1 , for all yogurts. The highest counts (6.86 log CFU mL-1 ) for L. delbrueckii subsp. bulgaricus was found for S2 yogurts made from whey protein hydrolysate (B3). Minor water-holding capacity was detected for YF-L811 (S1) yogurts. Yoflex Harmony 1.0 (S3) starter containing Lim. fermentum produced a distinctive volatile profile characterized by aldehydes with respect to yogurts prepared with S1, S2 and S4, which were characterized by ketones. CONCLUSION: Results indicate the usefulness of carrying out studies similar to the present one to select the most appropriate process conditions depending on the desired product. © 2022 Society of Chemical Industry.

Effect of the incorporation of ß-galactosidase in the GOS production during manufacture of soft cheese.

Galactooligosaccharides (GOS) are non-digestible oligosaccharides with recognized prebiotic role. The present study aims to evaluate a ß-galactosidase from K. lactis during soft cheese making and to analyse the impact on carbohydrates metabolism, proteolysis, and volatile compounds production, physicochemical and microbiological characteristics of the product. The enzyme was added to cheese milk (fluid milk plus whey powder) before (40 min.) or simultaneously of the starter addition (Ep and E treatments, respectively); cheese without enzyme addition was also made (C treatment). Also, we characterized fresh and soft commercial cheeses from the point of view of carbohydrate fraction, highlighting GOS, and organic acid profiles. The inclusion of the enzyme in soft cheese making produced a delay in reaching the target pH (~5.2). Carbohydrate fermentation profiles differed among treatments during cheese making and ripening. GOS were only detected in Ep and E cheeses (0.88 and 0.51 g/100 g, respectively). Lactose content was lower, and glucose and galactose levels were higher in E and Ep than C. No differences in physicochemical and microbial composition and organic acids profiles among samples were observed. Bioformation of volatile compounds belonging to the chemical families of aldehydes, ketones, alcohols, esters and acids, was not substantially affected by the modification in the carbohydrate profile. GOS were not detected in any of the commercial cheeses; great variations in the carbohydrate contents and organic acids were found. The results obtained demonstrate the feasibility of obtaining cheeses with GOS. Although the GOS values achieved are not adequate enough for the desired effect, the proposed technological approach turned out to be satisfying and original. Cheeses with prebiotic fiber are not still widespread in the market.

Evaluation of autochthonous cultures to improve the cheese flavor: A case study in hard cheese model.

The characterization of autochthonous cultures based on their contribution to cheese flavor is an additional selection criterion for their use in cheese making. The objective of the present work was to assess the ability of three strains of mesophilic lactobacilli: Lactobacillus casei 72 (Lc72), L. paracasei 90 (Lp90), and L. plantarum 91 (Lp91), one strain of thermophilic lactobacillus: L. helveticus 209 (Lh209), and the thermophilic-mesophilic combinations, to grow and produce aroma compounds in a hard cheese model. Microbiological counts, pH, and the profiles of carbohydrates, organic acids, and volatile compounds were analyzed during incubation for 14 days at 37 ℃. The population of mesophilic lactobacilli reached levels around 8.0 log CFU ml-1 at three days, but then decreased until ∼7.0 log CFU ml-1 toward 14 days. Thermophilic lactobacillus population reached and maintained levels around 7.7 log CFU ml-1 during incubation. Carbohydrates were absent in the hard cheese model, and so no change in the pH values and in the levels of lactic acid was detected. Mesophilic lactobacilli, inoculated individually or in association with Lh209, metabolized the citric acid and produced ethanoic acid. The profiles of volatile compounds of mesophilic lactobacilli (characterized mainly by butan-2-one, 3-hydroxybutan-2-one, 3-methylbutan-1-ol, hexan-1-ol, 2-phenylethanol, and ethanoic acid) were different from the profile of thermophilic lactobacillus Lh209 (characterized mainly by heptan-2-one, ethyl acetate, isoamyl hexanoate, pentan-1-ol, decanoic acid, and 2- and 3-methylbutanal). Cooperative effects in the production of compounds related to cheese flavor, such as 3-hydroxybutan-2-one, ethyl butanoate, ethanol, pentan-2-ol, hexan-1-ol, benzeneacetaldehyde, 2-phenylethanol, and heptanoic acid, were largely evidenced between Lh209 and Lp91; in a lesser extent, cooperative effects were also found for Lh209+Lp90 for the following compounds: 3-hydroxybutan-2-one, isoamyl acetate, and ethanoic acid. Of the mesophilic lactobacilli strains evaluated, Lp91 and Lp90 would be interesting candidates for its use as adjunct cultures in hard cheeses to improve and diversify the flavor.

Characterization of volatile compounds produced by Lactobacillus helveticus strains in a hard cheese model.

Starter cultures of Lactobacillus helveticus used in hard cooked cheeses play an important role in flavor development. In this work, we studied the capacity of three strains of L. helveticus, two autochthonous (Lh138 and Lh209) and one commercial (LhB02), to grow and to produce volatile compounds in a hard cheese extract. Bacterial counts, pH, profiles of organic acids, carbohydrates, and volatile compounds were analyzed during incubation of extracts for 14 days at 37 ℃. Lactobacilli populations were maintained at 106 CFU ml-1 for Lh138, while decreases of approx. 2 log orders were found for LhB02 and Lh209. Both Lh209 and LhB02 slightly increased the acetic acid content whereas mild increase in lactic acid was produced by Lh138. The patterns of volatiles were dependent on the strain which reflect their distinct enzymatic machineries: LhB02 and Lh209 produced a greater diversity of compounds, while Lh138 was the least producer strain. Extracts inoculated with LhB02 and Lh 209 were characterized by ketones, esters, alcohols, aldehydes, and acids, whereas in the extracts with Lh138 the main compounds belonged to aromatic, aldehydes, and ketones groups. Therefore, Lh209 and LhB02 could represent the best cheese starters to improve and intensify the flavor, and even a starter composed by combinations of LhB02 or Lh209 with Lh138 could also be a strategy to diversify cheese flavor.

Response surface methodology as a tool for modelling galacto-oligosaccharide production.

The experiments reported in this research paper describe the effects of ß-galactosidase enzyme dose and cheese whey amount, on the maximum concentration and yield of galacto-oligosaccahride (GOS) and reaction time. The experimental plan was based on central composite rotational design (CCRD) and modelled by response surface methodology (RSM). The results indicate that the proposed mathematical models could adequately describe the concentration and yield of GOS and the reaction time within the limits of the factors that are being investigated. The variance analysis shows high values of coefficients of determination (>0·97) while no significant lack of fit was evident. Hence, the models could be employed to select reaction conditions applied in the manufacture of products enriched in bioactive compounds with high value-added.

Cheese milk low homogenization enhanced early lipolysis and volatiles compounds production in hard cooked cheeses.

Homogenization applied to cheese milk has shown to increase lipolysis but its use is not spread as it can induce detrimental effects. The aim of this work was to assess the effect of low-pressure homogenization of the cream followed by pre-incubation of cheese milk on the composition, ripening index, lipolysis and volatile profiles of hard cooked cheeses. For that, control and experimental miniature Reggianito cheeses were made and analyzed during ripening (3, 45 and 90days). Homogenization had no impact on composition and proteolysis. An acceleration of the lipolysis reaction was clearly noticed in cheeses made with homogenized milk at the beginning of ripening, while both type of cheeses reached similar levels at 90days. We found the level of several compounds derived from fatty acid catabolism were noticeably influenced by the treatment applied: straight-chain aldehydes such as hexanal, heptanal and nonanal and methylketones from C5 to C9 were preferentially formed in experimental cheeses.

Influence of lactose hydrolysis on galacto-oligosaccharides, lactose, volatile profile and physicochemical parameters of different yogurt varieties.

BACKGROUND: Different types of reduced-lactose yogurt, obtained by lactose hydrolysis using ß-galactosidase enzyme, are commercially available. The breakdown of lactose modifies the carbohydrate profile, including the production of prebiotic galacto-oligosaccharides (GOS), which could affect the survival and activity of starter and probiotic cultures and the parameters of yogurt quality. The extension of these changes is dependent on the yogurt matrix composition. This study aimed to evaluate the influence of lactose hydrolysis on GOS, lactose, volatile profile and physicochemical parameters of different yogurt varieties during storage. RESULTS: The presence of ß-galactosidase enzyme did not affect either the global composition or the survival of cultures. Overall, the hydrolyzed products had lower acidity than traditional ones. GOS were found at similar levels in fresh hydrolyzed yogurts, whereas in traditional yogurts they were not detected. The proportion of ketones, acids and aldehydes seems to be more dependent on yogurt variety than on addition of the enzyme. Likewise, the storage period affected the volatile fraction to different degree; the increase in acid compounds was more pronounced in hydrolyzed than in traditional yogurts. CONCLUSION: This work shows that it is possible to obtain different varieties of reduced-lactose yogurt, some of them with additional benefits to health such as reduced fat, reduced calories, added with probiotic/inulin and enriched in GOS, with similar characteristics to traditional products. © 2016 Society of Chemical Industry.

Influence of autochthonous adjunct cultures on ripening parameters of Argentinean goat's milk cheeses.

BACKGROUND: Argentinean semi-hard goat's cheeses manufactured with and without the addition of autochthonous adjunct cultures of Lactobacillus plantarum ETC17, Lactobacillus rhamnosus ETC14 and Enterococcus faecium ETC3 were analysed to evaluate the effect of these strains on ripening parameters. RESULTS: Gross composition was similar among cheeses. Microbiological analysis indicated that lactic acid bacteria added to cheeses reached high levels. None of the strains assayed affected the primary proteolysis. Overall, E. faecium had a clearer effect on the peptide and lipolysis profiles of cheeses. Analysis of the volatile fraction of cheeses indicated that the levels of several compounds involved in the overall flavour of goat's cheeses were affected by the presence of E. faecium. This could explain the differences detected in the global perception of cheeses made with this strain compared with control cheeses. CONCLUSION: The present work represents a first contribution to knowledge of the ripening process of Argentinean goat's cheeses made with the addition of autochthonous adjunct cultures. The results suggest that E. faecium ETC3 showed a significant effect during ripening, which was reflected both in the profiles of proteolysis, lipolysis and volatile compounds and in the global sensory perception of cheeses.

Composition and volatile profiles of commercial Argentinean blue cheeses.

BACKGROUND: A first approach was made to acquire knowledge of the global composition, proteolysis, lipolysis and volatile profile of Argentinean blue cheeses. A total of 20 samples belonging to six leading commercial brands were analysed. A comparison of the results with bibliographical data on other blue cheeses was performed and the variability among and within dairy plants was also evaluated. RESULTS: Values of global composition were intermediate in relation to those reported for mould-ripened cheeses. Levels of proteolysis and lipolysis were lower than those of other blue cheeses. Volatile compound profiles were characterised mainly by short-chain fatty acids, methyl ketones and secondary alcohols, as reported previously for blue cheeses. Wide-ranging values of physicochemical parameters, lipolysis and proteolysis levels as well as volatile compound areas of cheeses produced by each dairy plant were observed. Owing to this high variability in the chemistry and volatile profile of cheeses, principal component analysis of the data did not show groupings by commercial brands. CONCLUSION: On the whole, Argentinean blue cheeses were characterised both by gross intermediate composition values and by proteolysis and lipolysis levels lower than those of blue cheeses of different origins. A typical volatile compound profile of blue cheeses was observed. The high variability found within each commercial brand could reflect the lack of standardisation of the technological processes used in blue cheese manufacturing in Argentina.