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.

Root phospholipids in Azospirillum-inoculated wheat seedlings exposed to water stress.

Azospirillum-plant association is accompanied by biochemical changes in roots which, in turn, promote plant-growth and tolerance to water stress. To shed light on the possible factors underlying these effects, roots from Azospirillum brasilense Sp245-inoculated Triticum aestivum seedlings growing in darkness under osmotic stress were analyzed for phospholipid (PL) composition, fatty acid (FA) distribution profiles and degree of unsaturation of the major PL classes. Azospirillum inoculation diminished ion leakage and increased 2,3,5-tripheniltetrazolium reducing ability in roots of well irrigated and water-stressed wheat seedlings. Total root PL content remained unaltered in all treatments. Six PL classes were detected, phosphatidylcholine (PC) and phosphatidylethanolamine (PE) comprising over 80% of the total. While water stress increased PC content and diminished that of PE, none of these changes were observed either under Azospirillum inoculation alone or when both treatments were combined. The major FAs found in both PC and PE were 16:0, 18:0, 18:1, 18:2, and 18:3. Higher PC and lower PE unsaturation than in well irrigated controls were observed in roots from Azospirillum-inoculated, water-stressed seedlings. Azospirillum inoculation could contribute to protect wheat seedlings from water stress through changes in the FA distribution profiles of PC and PE major root phospholipids.

Proteolysis on Reggianito Argentino cheeses manufactured with natural whey cultures and selected strains of Lactobacillus helveticus.

Reggianito Argentino cheese is traditionally manufactured with whey starter cultures that provide typical and intense flavor but can cause poor quality standardization. In this study, the influence of natural and selected starters on Reggianito Argentino cheese proteolysis was investigated. Cheeses were manufactured with three strains of Lactobacillus helveticus (SF133, SF138 and SF209) cultured individually in sterile whey and used as single or mixed starters. Control cheeses were made with natural whey starter culture. Cheeses were analyzed to determine gross composition, as well as total thermophilic lactic flora. Proteolysis was assessed by N fractions, electrophoresis and liquid chromatography. Gross composition of the cheeses did not significantly differ, while viable starter cell counts were lower for cheeses made with strain SF209 alone or combined with other strains. Soluble N at pH 4.6 was the same for cheeses made with natural or selected starters, but soluble N in 12% trichloroacetic acid and 2.5% phosphotungstic acid was significantly higher in cheeses made with starters containing strain SF209. Nitrogen fractions results indicated that natural whey starter cultures could be replaced by several starters composed of the selected strains without significant changes to proteolysis patterns. Starter cultures prepared only with SF209 or with the three selected L. helveticus strains produced cheese products with significantly more proteolysis than control cheeses. Chromatographic profiles analyzed by principal components showed that three main peaks on chromatograms, presumptively identified as Tyr, Phe, and Trp, explained most of variability. Principal component scores indicated that cheese samples were grouped by ripening time, which was confirmed by linear discriminant analysis. On the contrary, samples did not cluster by Lactobacillus strain or type of starter.

Influence of milk-clotting enzyme concentration on the alphas1-casein hydrolysis during soft cheeses ripening.

We studied the influence of the dose of milk-clotting enzyme on alphas1-CN degradation, soluble nitrogen production, and sensory profile for an Argentinean soft cheese: Cremoso Argentino. Five different types of cheeses were produced: 1) control cheeses with normal technology, 2) cheeses with inactivated milk-clotting enzyme, 3) cheeses with inactivated milk-clotting enzyme, without starter (acidified with glucono delta lactone), 4) cheeses with a half dose of milk-clotting enzyme, and 5) cheeses with a double dose of milk-clotting enzyme. Proteolysis was assessed by isoelectric focusing electrophoresis of the insoluble fraction at pH 4.6, followed by densitometric quantification. Soluble nitrogen at pH 4.6, expressed as a percentage of total nitrogen and defined as ripening index was also performed. A sensorial panel evaluated the cheeses at the end of ripening. The hydrolysis level of alphas1-CN depended on the milk-clotting enzyme dose used in cheese making. Cheeses without active coagulant did not show degradation at the end of ripening, while cheeses with half and whole doses showed proportional degradations to coagulant dose. Cheese with a double dose of coagulant did not show higher alphas1-CN hydrolysis than normal cheese. No difference was found between cheeses with and without microbiological starter, indicating that the selected culture, composed of thermophilic strains, was unable to attack the whole casein. A high linear correlation was found between ripening index and the relation Sensorial characteristics of cheeses agree with objective analysis. Cheeses without active coagulant were hard and crumbly, while cheeses with normal dose were soft and creamy.

[Production of a concentrate of Mucor bacilliformis acid protease]. / Produccion de un concentrado de proteasa ácida de Mucor bacilliformis.

A concentrate of milk-clotting enzyme was produced by culture of Mucor bacilliformis on wheat bran medium moistened to 120% water on dry bases with HC1 2 N solution. The wheat bran was autoclaved, spread on trays and inoculated with 5.10(6) spore/gr of dry bran. After 10 days of culture at 21 degrees C, the enzyme produced was extracted with water and adjusted to pH 4.4. The precipitation was performed with ethanol. The precipitate was dissolved in HCl solution (pH 4.5) and it was concentrated by dialysis against polyethylene glycol 20.000. The enzyme solution had a specific activity of 1123 units/mg. and it was tested in the elaboration of cream cheese.

Produccion de un concentrado de proteasa acida de Mucor bacilliformis. / Production of an acid protease concentrate from Mucor bacilliformis

Se obtuvo un concentrado de enzimas congulantes de leche por cultivo de Mucor bacilliformis en afrecho de trigo humectado a 120% de agua sobre base seca con solucion de HC1 0,2 N. El afrecho esterilizado fue dispuesto en bandejas, e inoculado con 5.10 (6) esporos/gr de afrecho seco. Luego de 10 dias de incubacion se extrajo el afrecho enmohecido con agua, se ajusto el pH a 4,4 y se precipito el sobrenadante limpido con etanol. El precipitado fue disuelto en agua clorhidrica (pH 4,5) y concentrado por dialisis contra polietilenglicol 20000. La solucion con una actividad especifica de 1128 U/mg, se utilizo en dos ensayos de elaboracion de queso tipo cremoso

Produccion de un concentrado de proteasa ácida de Mucor bacilliformis. / [Production of a concentrate of Mucor bacilliformis acid protease]

A concentrate of milk-clotting enzyme was produced by culture of Mucor bacilliformis on wheat bran medium moistened to 120

water on dry bases with HC1 2 N solution. The wheat bran was autoclaved, spread on trays and inoculated with 5.10(6) spore/gr of dry bran. After 10 days of culture at 21 degrees C, the enzyme produced was extracted with water and adjusted to pH 4.4. The precipitation was performed with ethanol. The precipitate was dissolved in HCl solution (pH 4.5) and it was concentrated by dialysis against polyethylene glycol 20.000. The enzyme solution had a specific activity of 1123 units/mg. and it was tested in the elaboration of cream cheese.

Produccion de un concentrado de proteasa acida de Mucor bacilliformis. / Production of an acid protease concentrate from Mucor bacilliformis

Se obtuvo un concentrado de enzimas congulantes de leche por cultivo de Mucor bacilliformis en afrecho de trigo humectado a 120% de agua sobre base seca con solucion de HC1 0,2 N. El afrecho esterilizado fue dispuesto en bandejas, e inoculado con 5.10 (6) esporos/gr de afrecho seco. Luego de 10 dias de incubacion se extrajo el afrecho enmohecido con agua, se ajusto el pH a 4,4 y se precipito el sobrenadante limpido con etanol. El precipitado fue disuelto en agua clorhidrica (pH 4,5) y concentrado por dialisis contra polietilenglicol 20000. La solucion con una actividad especifica de 1128 U/mg, se utilizo en dos ensayos de elaboracion de queso tipo cremoso