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  • Effects of different ripening procedures on the final characteristics
    Publication . Freitas, A. C.; Malcata, F. X.
    Picante da Beira Baixa (or Picante) cheese is a hard, piquant, salted traditional cheese manufactured in Portugal from raw sheep’s and goat’s milks. The purpose of this work was to quantitatively assess the influence of various ripening procedures on the final characteristics of Picante cheese. Two alternative ripening protocols were considered, the traditional one and another with controlled environmental conditions via use of maturation chambers set at different preselected temperatures. The experimental cheeses were characterised in terms of microbiological, physicochemical, biochemical, sensorial and textural properties. Ripening time and temperature were statistically significant parameters for all microflora. The two ripening methods led to statistically significant differences in all physicochemical and biochemical parameters, especially the moisture content and the soluble nitrogen fractions (i.e. water loss was slower and proteolysis was faster in cheeses ripened via the traditional method). Differences in microbiological, physicochemical and biochemical properties were probables implicated in differences in textural and sensorial properties, especially cheese hardness and flavour. It was concluded that the standard ripening method was closest to the traditional one in terms of final cheese characteristics when the ripening temperature was above 11.5 7C.
  • Caseins as source of bioactive peptides
    Publication . Silva, Sofia V.; Malcata, F. Xavier
    Biologically active peptides are of particular interest in food science and nutrition because they have been shown to play physiological roles, including opioid-like features, as well as immunostimulating and anti-hypertensive activities, and ability to enhance calcium absorption. Hidden or inactive in the amino-acid sequence of dairy proteins, they can be released or activated in vivo during gastrointestinal digestion, or upstream during food processing via specific, enzyme-mediated proteolysis. Caseins, in either milk or dairy products (e.g. cheese), are important sources of those peptides; their biological significance, their impact on human health and the manufacture of novel functional food ingredients therefrom have been subject to intensive research, which will be briefly presented and critically discussed in this review.
  • Role of dominant microflora of Picante cheese on proteolysis and lipolysis
    Publication . Freitas, A. Cristina; Pintado, Ana I. E.; Pintado, Manuela; Malcata, F. Xavier
    Four species of bacteria (Enterococcus faecium and E. faecalis, Lactobacillus plantarum and Lb. paracasei) and three species of yeasts (Debaryomyces hansenii, Yarrowia lipolytica and Cryptococcus laurentii) isolated from Picante cheese were assayed for proteolytic and lipolytic activities. The milk type (caprine or ovine), the ripening time (0–65 d) and the concentration of NaCl (0–14% (w/v)) have been studied in terms of their effects upon in vitro curdled milk. Proteolytic and peptidolytic activities were demonstrated to be high for Y. lipolytica, and at much lower levels for the other strains. Milk type, ripening time and content of NaCl appeared to be statistically significant processing factors in terms of proteolysis. Clear lipolytic activity was detected for Y. lipolytica, but release of free fatty acids to lesser extents was observed for the other strains under study. Ripening time was statistically significant for lipolysis but milk type was not. Lipolytic activities were strongly affected by NaCl content and the extent of fat hydrolysis was affected by the increase of NaCl from 0 to 7% (w/v) more than by change from 7 to 14% (w/v). In view of the experimental results, a mixed-strain starter for Picante cheese including Lb. plantarum, E. faecium (or E. faecalis) and D. hansenii (and/or Y. lipolytica) is of potential interest.
  • Caprine cheese with probiotic strains: the effects of ripening temperature and relative humidity on proteolysis and lipolysis
    Publication . Gomes, Ana M. P.; Silva, M. Luz P. C.; Malcata, F. Xavier
    The effects of ripening temperature, relative humidity and time on chemical and textural characteristics of a ‘probiotic’ goat’s milk cheese were examined. The experimental layout followed a 23 factorial design, with all possible combinations of 5 7C and 10 7C (ripening temperature), 85% and 95% (ripening relative humidity) and 1 day and 70 days (ripening time). All proteolytic indices measured (water-soluble nitrogen, trichloroacetic acid-soluble nitrogen and phosphotungstic acid-soluble nitrogen) were enhanced with increased ripening temperature to a greater extent than with increased ripening relative humidity; the increase in phosphotungstic acid-soluble nitrogen was the most significant. Free fatty acid concentrations in cheeses were not influenced by ripening relative humidity but increased with ripening temperature and time. A higher ripening temperature and a lower relative humidity gave rise to firmer cheeses. Postulated empirical models have provided a good fit to the experimental data set generated; such models were able to predict a decrease of 25 days in ripening time with no impairment of either proteolytic or lipolytic indices if a cheese were to be ripened at 10 7C (rather than 5 7C) and 95% relative humidity.
  • Degradation of caseins from milk of different species by extracts of Centaurea calcitrapa
    Publication . Tavaria, Freni K.; Sousa, M. José; Domingos, Ana; Malcata, F. Xavier; Brodelius, Peter; Clemente, Alda; Pais, M. Salomé
    Two purified aqueous extracts of Centaurea calcitrapa, a plant from the Compositae family, were shown to degrade bovine, ovine, and caprine milk caseinates. The corresponding hydrolysis patterns were monitored by urea-polyacrylamide gel electrophoresis and compared with those of a commercial mixture of chymosin and pepsin. The plant proteases degraded both Rs- and â-caseins more extensively than the commercial rennet, thus yielding proteolytic patterns where different specificities toward such substrates are apparent. The animal rennet was found to display the greatest clotting power per milligram of protein but the least specificity toward ovine and caprine caseins. It is, therefore, suggested that the said plant extracts be used as an alternative to commercial animal rennets especially in the manufacture of caprine and ovine milk cheeses.
  • Assessment of proteolysis and lipolysis in Serra cheese: effects of axial cheese location, ripening time and lactation season
    Publication . Macedo, A. C.; Costa, M. L.; Malcata, F. X.
    The degrees of proteolysis and Iipolysis in Serra cheese were measured in cheese samples obtained from experiments laid out as a three-way facto rial design replicated twice. The independent variables studied were the location within the cheese (from the center to the surface), the position in the lactation season (from October to June) and the ripening time (from 0 to 35 days). Ali three variables had statistically significant effects on the concentration of water soluble peptides (WSP), but only the ripening time and the period within the lactation season affected the fat acidity (FA). The extents (alter the given ripening period) and the rates (averaged over the ripening period) of generation of WSP and FA were highest for spring and lowest for autumn. The lowest values for both the proteolysis extent and the proteolysis rate were obtained for the rind. The rates of proteolysis and lipolysis tended to decrease with ripening time; most of the lipolysis occurred during the first week, but proteolysis was still in progress by 35 days of ripening.
  • Sweet whey cheese matrices inoculated with the probiotic strain Lactobacillus paracasei LAFTI® L26
    Publication . Madureira, Ana Raquel; Soares, José Carvalho; Pintado, M. E.; Gomes, Ana M. P.; Freitas, A. C.; Malcata, F. Xavier
    Consumption of dairy products containing viable probiotic strains has increased dramatically in recent years, owing to general health claims associated therewith. This trend has boosted diversification of the portfolio of said products, including whey cheese matrices. However, taking into account the relatively poor organoleptic and textural features of these matrices, improvement is in order via incorporation of selected additives, provided that viability of the strains is duly assayed. Lactobacillus paracasei LAFTI® L26 was accordingly incorporated into whey protein solid matrices, in the presence of several additives aimed at enhancing their organoleptic appeal and textural performance. These matrices were produced from a combination of either ovine or bovine whey (or a mixture thereof) with ovine milk, and were inoculated at 10% (v/v) with the probiotic strain. Sugar, sugar and aloe vera, sugar and chocolate, and sugar and jam were further added, and the resulting products were then stored at 7 ◦C for 21 d. In general, viable cell numbers remained high in all experimental matrices throughout storage. Despite the observed low extents of breakdown, proteolytic activities by the end of storage were higher in matrices containing jam. Furthermore, L. paracasei partially converted lactose into lactic acid in these matrices. Additives enhanced the organoleptic features of whey cheeses, and produced different textural patterns. The higher sensory scores were attained by matrices containing sugar: sugar and aloe vera received the best scores by 3 d of storage, but these scores decreased as storage time elapsed.
  • Influence of native lactic acid bacteria on the microbiological, biochemical and sensory profiles of Serra da Estrela cheese
    Publication . Macedo, Angela C.; Tavares, Tânia G.; Malcata, F. Xavier
    Cheesemaking from batches of raw ewe’s milk was carried out via inoculation with wild strains of Lactococcus lactis subsp. lactis ESB110019 and Lactobacillus plantarum ESB5004 independently, or combined with each other. Those two strains had been isolated from the native microflora of typical Serra da Estrela cheese. One control batch was processed in parallel without addition of any starter. The evolution in viable counts of the main micro-organisms (viz. lactic acid bacteria, Enterobacteriaceae, staphylococci and yeasts), as well as in secondary proteolysis (WSN, 2% TCASN, 12% TCASN and 5% PTASN), was monitored throughout ripening time (over a 63-day period) in cheeses from each batch. The sensory features of the fully ripened cheeses were also assessed. Cheeses manufactured with starter showed significantly lower levels of viable Enterobacteriaceae than those manufactured without starter; viable counts of enterococci and staphylococci did significantly increase after addition of L. lactis or Lb. plantarum, respectively. Proteolysis in terms of WSN and 5% PTASN was not significantly affected by the lactic acid bacteria tested when compared to the control, but L. lactis played a significant role toward increasing the 2% TCASN content of cheeses; both strains led to a statistically significant increase of the 12% TCASN. The scores for flavor and texture of the control cheeses were somewhat above those for the experimental cheeses manufactured with starter.
  • Storage and lyophilization effects of extracts of Cynara cardunculus on the degradation of ovine and caprine caseins
    Publication . Tavaria, Freni K.; Sousa, M. José; Malcata, F. Xavier
    Clotting and proteolytic activities are important parameters when evaluating rennets for cheesemaking. Both these activities were determined for extracts of the plant Cynara cardunculus in fresh form and after lyophilization followed by reconstitution, either in water or in citrate buffer (pH 5.4) and stored for up to 4 weeks at 4°C. The patterns of degradation of ovine and caprine caseins were followed by urea polyacrylamide gel electrophoresis in attempts to qualitatively differentiate the activity of the enzyme extracts as storage time elapsed. Storage at 4°C significantly decreased the clotting power of the extracts but lyophilization retarded this decrease; β- and αs-casein breakdown generally increased with storage time, via patterns that depend on caseinate type and extract used, but lyophilized extracts reconstituted in citrate buffer were significantly less proteolytic than the other extracts. Therefore, it is suggested that lyophilized extracts (reconstituted with citrate buffer) of flowers of C. cardunculus be used rather than fresh extracts.
  • How milk type, coagulant, salting procedure and ripening time affect the profile of free amino acids in Picante da Beira Baixa cheese
    Publication . Freitas, A. C.; Fresno, J. María; Prieto, Bernardo; Franco, Imaculada; Malcata, F. Xavier; Carballo, Javier
    The concentration of total free amino acids (FAA) in Picante cheese increased with ripening time irrespective of the particular protocol used for manufacture (ie ratio of caprine to ovine milks, animal or plant rennet and number of salting steps). The experimental cheeses manufactured with 20% (v/v) caprine milk, coagulated with animal rennet and salted only once exhibited the highest content of total FAA by 120 days of ripening. All four manufacture parameters were statistically significant on the 0.5% level of significance in terms of total concentration of FAA. The dominating free amino acids present in the various experimental cheeses throughout the ripening period were valine, leucine and phenylalanine, each one representing more than 10% (w/w) of the total concentration of FAA. All four manufacture parameters were, in general, statistically significant with respect to the content of every single FAA, with particular emphasis on salting and ripening time.