Erica R. Hynes

Reggianito Argentino cheese: influence of Lactobacillus helveticus strains isolated from natural whey cultures on cheese making and ripening processes

Abstract Reggianito Argentino cheeses were manufactured with three defined single strains Lactobacillus helveticus cultured in sterile whey, and one “natural” whey starter. Gross composition of cheeses did not significantly differ, and viable starter cell counts were similar for all cheeses. Soluble nitrogen at pH 4.6 was also alike for cheeses made with natural or selected starters, but soluble nitrogen in trichloroacetic acid 12% and phosphotungstic acid 2.5% showed significant differences. Electrophoretograms showed that γ casein bands increased during ripening, while β casein band decreased; α s1 casein was also cleaved to α s1 -I. Acid degree values of fat for control and experimental 0-day old cheeses were slightly different, but were similar at 90 and 180 days of ripening. All cheeses were good quality Reggianito, but control and experimental samples differed in aroma and texture. Natural whey starter replacement by selected single starter of Lactobacillus did not alter cheese making and primary proteolysis or lipolysis, but it modified secondary proteolysis and sensory characteristics.

Influence of milk pretreatment on production of free fatty acids and volatile compounds in hard cheeses: Heat treatment and mechanical agitation

This work aimed to identify technological steps that can increase fat hydrolysis and volatile compounds production in hard cheeses; these biochemical events have been related with improved piquant taste and development of genuine flavor during cheese ripening. For that purpose, 2 different pretreatments of cheese milk were tested: heat treatment and mechanical agitation. Both factors were assayed at 2 levels: milk was either batch pasteurized or nonthermally treated, and mechanical agitation was either applied or not applied. For all combinations, hard cheeses (Reggianito type) were produced in a pilot plant and ripened for 90 d. In all cheeses the degree of lipolysis, assessed by gas chromatography, increased similarly during ripening. However, the proportion of short-chain fatty acids was higher in the cheeses made with unpasteurized milk, suggesting a higher activity of lipases with positional specificity toward the sn-3 position of the triglyceride, among which milk lipoprotein lipase is found. Similar results were found for most of the volatile compounds, determined by solid-phase microextraction-gas chromatography flame-ionization detector/mass spectrometry, which constitute the groups of ketones, alcohols, esters, and the group of acids. On the contrary, no effect of mechanical agitation was observed, although some interactions between factors were found. In the conditions of the study, results suggest that heat treatment had a higher effect on cheese lipolysis and volatile compounds production than partial destabilization of the fat emulsion produced by the agitation method applied.

Multivariate analysis of proteolysis patterns differentiated the impact of six strains of probiotic bacteria on a semi-hard cheese.

The individual contribution of 6 strains of probiotic bacteria (3 of Lactobacillus acidophilus and 3 of the Lactobacillus casei group) to proteolysis patterns in a semi-hard cheese was assessed. Control cheeses (without probiotics) and 2 types of experimental cheeses (with the addition of probiotics either directly to milk or by a 2-step fermentation method) were manufactured. Cheeses containing Lb. acidophilus showed the most extensive peptidolysis, which was evidenced by changes in the peptide profiles and a noticeable increase of free amino acids compared with control cheeses. The strains of the Lb. casei group showed a lower contribution to cheese peptidolysis, which consisted mainly of free amino acid increase. Two-step fermentation improved peptidolytic activity for only one of the cultures of Lb. acidophilus tested. The addition of Lb. acidophilus strains into cheese may be suitable not only for their beneficial health effect but also for their influence on secondary proteolysis, consistent with acceleration of ripening and improved flavor formation.

Impact of Chymosin- and Plasmin-Mediated Primary Proteolysis on the Growth and Biochemical Activities of Lactobacilli in Miniature Cheddar-Type Cheeses

Strongly proteolytic starters seem to improve the growth of nonstarter lactobacilli during cheese ripening, but no information is available on the impact of the nonmicrobial proteases usually active in cheese on their development. In the current study, the influence of chymosin- and plasmin-mediated proteolysis on the growth and biochemical activities of lactobacilli during ripening of miniature Cheddar-type cheeses, manufactured under controlled microbiological conditions, was studied. Two experiments were performed; in the first, residual chymosin activity was inhibited by the addition of pepstatin, and in the second, plasmin activity was increased by adding more enzyme, obtained in vitro through the activation of plasminogen induced by urokinase. Cheeses with or without a Lactobacillus plantarum I91 adjunct culture and with or without added pepstatin or plasmin solution were manufactured and ripened for 60 d. The addition of the adjunct culture resulted in enhancement of secondary proteolysis, evidenced by an increase in the total content of free amino acids (FAA) and modifications of the individual FAA profiles. Reduction in residual chymosin activity caused a decrease in primary and secondary proteolysis, characterized by the absence of alpha(s1)-casein hydrolysis and reduced production of peptides and FAA, respectively. The increase in plasmin activity accelerated primary proteolysis but no enhancement of secondary proteolysis was observed. Chymosin- and plasmin-mediated proteolysis did not influence the growth and biochemical activities of adventitious or adjunct lactobacilli, indicating that it is not a limiting factor for the development and proteolytic-peptidolytic activities of lactobacilli in the cheese model studied.

Aminotransferase and glutamate dehydrogenase activities in lactobacilli and streptococci

Aminotransferases and glutamate dehydrogenase are two main types of enzymes involved in the initial steps of amino acid catabolism, which plays a key role in the cheese flavor development. In the present work, glutamate dehydrogenase and aminotransferase activities were screened in twenty one strains of lactic acid bacteria of dairy interest, either cheese-isolated or commercial starters, including fifteen mesophilic lactobacilli, four thermophilic lactobacilli, and two streptococci. The strains of Streptococcus thermophilus showed the highest glutamate dehydrogenase activity, which was significantly elevated compared with the lactobacilli. Aspartate aminotransferase prevailed in most strains tested, while the levels and specificity of other aminotransferases were highly strain- and species-dependent. The knowledge of enzymatic profiles of these starter and cheese-isolated cultures is helpful in proposing appropriate combinations of strains for improved or increased cheese flavor.

Influence of chymosin type and curd scalding temperature on proteolysis of hard cooked cheeses.

In this work, we studied the influence of the type of coagulant enzyme and the curd scalding temperature on the proteolysis and residual coagulant and plasmin activities of a cooked cheese, Reggianito, in the interest of reducing ripening time. A two-factor experimental design was applied in two levels: type of coagulant enzyme, bovine chymosin or camel chymosin, and curd scalding temperature, 50 or 56 °C. The experimental treatments were applied in Reggianito cheese making experiments, and the samples were ripened for 90 d at 12 °C. Scalding temperature influenced residual coagulant activity; the cheeses cooked at 50 °C had significantly higher activity than those treated at 56 °C. In contrast, scalding temperature did not modify plasmin activity. Proteolysis was primarily affected by curd cooking temperature because chymosin-mediated hydrolysis of αs1 casein was slower in cheeses treated at 56 °C. Additionally, the nitrogen content in the cheese soluble fractions was consistently lower in the cheeses scalded at 56 °C than those cooked at 50 °C. A significant influence of the type of coagulant enzyme was observed, especially in the nitrogen fractions and peptide profiles, which demonstrated that camel chymosin was slightly less proteolytic; however, these differences were lower than those caused by the scalding temperature.

Growth, survival, and peptidolytic activity of Lactobacillus plantarum I91 in a hard-cheese model

In this work, we studied the growth, survival, and peptidolytic activity of Lactobacillus plantarum I91 in a hard-cheese model consisting of a sterile extract of Reggianito cheese. To assess the influence of the primary starter and initial proteolysis level on these parameters, we prepared the extracts with cheeses that were produced using 2 different starter strains of Lactobacillus helveticus 138 or 209 (Lh138 or Lh209) at 3 ripening times: 3, 90, and 180 d. The experimental extracts were inoculated with Lb. plantarum I91; the control extracts were not inoculated and the blank extracts were heat-treated to inactivate enzymes and were not inoculated. All extracts were incubated at 34°C for 21 d, and then the pH, microbiological counts, and proteolysis profiles were determined. The basal proteolysis profiles in the extracts of young cheeses made with either strain tested were similar, but many differences between the proteolysis profiles of the extracts of the Lh138 and Lh209 cheeses were found when riper cheeses were used. The pH values in the blank and control extracts did not change, and no microbial growth was detected. In contrast, the pH value in experimental extracts decreased, and this decrease was more pronounced in extracts obtained from either of the young cheeses and from the Lh209 cheese at any stage of ripening. Lactobacillus plantarum I91 grew up to 8 log during the first days of incubation in all of the extracts, but then the number of viable cells decreased, the extent of which depended on the starter strain and the age of the cheese used for the extract. The decrease in the counts of Lb. plantarum I91 was observed mainly in the extracts in which the pH had diminished the most. In addition, the extracts that best supported the viability of Lb. plantarum I91 during incubation had the highest free amino acids content. The effect of Lb. plantarum I91 on the proteolysis profile of the extracts was marginal. Significant changes in the content of free amino acids suggested that the catabolism of free amino acids by Lb. plantarum I91 prevailed in a weakly proteolyzed medium, whereas the release of amino acids due to peptidolysis overcame their catabolism in a medium with high levels of free amino acids. Lactobacillus plantarum I91 was able to use energy sources other than lactose to support its growth because equivalent numbers of cells were observed in extracts containing residual amounts of lactose and in lactose-depleted extracts. The contribution of Lb. plantarum I91 to hard-cooked cheese peptidolysis was negligible compared with that of the starter strain; however, its ability to transform amino acids is a promising feature of this strain.

Bioactive compounds delivery using nanotechnology: design and applications in dairy food

The novel properties of nanomaterials offer many new opportunities for the food industry. Different types of nanostructures can be incorporated into food in order to introduce new functionalities. These include; nanoliposomes, nanoemulsions, and nanoparticles. The sizes, shape, and internal structure of the particles vary considerably depending on the method and materials used to fabricate them. Bioactive compounds such as vitamins, antioxidants, and lipids can be protected using nanotechnology, even enhancing bioactivity and functionality. The design and application of nanoderived assemblies as tools for improved delivery and bioavailability of bioactive nutrients is promising. In this sense, success depends on scientific knowledge about degradation mechanisms of nutrients and major factors affecting them. Besides, it is necessary to review information in order to decide which delivery system fix with the desired application. In particular, nanotechnology has the potential to revolutionize dairy technology in the coming years.

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.

Short communication: A new minicurd model system for hard cooked cheeses

The aim of this study was to propose and validate a new minicurd model of young hard cheese. Curd particles and whey obtained from conventional cheese making of Reggianito Argentino were separated and frozen. Then, both fractions were thawed and the mixture of whey and curds was reconstituted, from which minicurds were made on the laboratory scale. Repeatability and the effect of freezing on minicurd composition were investigated by assessing pH, protein and moisture contents, sodium chloride content, and total thermophilic lactic flora counts. Good repeatability was achieved, and no significant differences were found between minicurds made from fresh compared with frozen materials. Composition of the minicurd was appropriate for modeling Reggianito Argentino cheese.