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.

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.

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.