Proteolysis of low-moisture Mozzarella cheese as affected by substitution of NaCl with KCl

The proteolytic and ACE inhibitory activities of low-moisture Mozzarella cheese (LMMC) as affected by partial substitution of NaCl with KCl were investigated. Experimental LMMC were made and salted with 4 salt mixtures: NaCl only (control), 3NaCl:1KCl, 1NaCl:1KCl, and 1NaCl:3KCl, and then proteolytic activity and angiotensin-converting enzyme inhibitory activity were determined. Salt treatment significantly affected angiotensin-converting enzyme inhibitory activity and phosphotungstic acid-soluble N of LMMC during storage. Water-soluble N, trichloroacetic acid-soluble N, lactic acid bacteria population, and total free amino acids were unaffected during storage. Nonetheless, water-soluble N and trichloroacetic acid-soluble N increased significantly during storage within a salt treatment. Peptide profiles and urea-PAGE gels did not differ between experimental cheeses at the same storage time.     

Incorporation of Lactobacillus casei in Iranian ultrafiltered Feta cheese made by partial replacement of NaCl with KCl

Probiotic Iranian ultrafiltered Feta cheese was produced from ultrafiltration of milk with a volumetric concentration factor of 4.5:1. The heat-treated retentates were inoculated with 10(7) cfu of Lactobacillus casei LAFTI L26/mL. A mesophilic-thermophilic mixed culture of Lactococcus lactis ssp. lactis, Lactococcus lactis ssp. cremoris, and Streptococcus thermophilus was also used. Three percent (wt/wt) salt with different ratios of NaCl:KCl (100% NaCl, 50% NaCl:50% KCl, 75% NaCl:25% KCl, and 25% NaCl:75% KCl) were used in cheese formulation. The viability of L. casei was determined in treatments during the ripening period (90d at 5°C) within 15-d intervals. The pH, titratable acidity, and redox potential changes were monitored throughout the mentioned period. The mean pH drop rate, mean acidity increase rate, and mean redox potential increase rate were calculated at the end of the storage period. Also, total nitrogen, water-soluble nitrogen, lactic acid, and acetic acid concentrations, and syneresis and sensory characteristics of the product were measured during the mentioned period every 30d. The maximum viability of L. casei was observed within d 15 to 30 of the ripening period in the treatment containing the lowest amount of sodium. Addition of KCl enhanced syneresis. Cheeses with NaCl alone and with only 25% replacement by KCl have the highest sensory acceptability.     

Effect of addition of Versagel on microbial, chemical, and physical properties of low-fat yogurt

The objective of this study was to examine the effect of Versagel on the growth and proteolytic activity of Streptococcus thermophilus 1275 and Lactobacillus delbrueckii ssp. bulgaricus 1368 and angiotensin-I converting enzyme inhibitory activity of the peptides generated thereby as well as on the physical properties of low-fat yogurt during a storage period of 28 d at 4 degrees C. Three different types of low-fat yogurts, YV0, YV1, and YV2, were prepared using Versagel as a fat replacer. The fermentation time of the low-fat yogurts containing Versagel was less than that of the control yogurt (YV0). The starter cultures maintained their viability (8.68 to 8.81 log CFU/g of S. thermophilus and 8.51 to 8.81 log CFU/g of L. delbrueckii ssp. bulgaricus) in all the yogurts throughout the storage period. There was some decrease in the pH of the yogurts during storage and an increase in the concentration of lactic acid. However, the proteolytic and ACE-inhibitory potential of the starter cultures was suppressed in the presence of Versagel. On the other hand, the addition of Versagel had a positive impact on the physical properties of the low-fat yogurt, namely, spontaneous whey separation, firmness, and pseudoplastic properties.     

The effect of substituting NaCl with KCl on Nabulsi cheese: chemical composition, total viable count, and texture profile

The effect of substituting NaCl with KCl on Nabulsi cheese characteristics was investigated. Nabulsi cheese was made and stored in 4 different brine solutions at 18%, including NaCl only (A; control); 3NaCl:1KCl (wt/wt; B); 1NaCl:1KCl (wt/wt; C); and 1NaCl:3KCl (wt/wt; D). Chemical composition, proteolysis, total viable count, and texture profile analysis were assessed at monthly intervals for 5 mo. No significant effect was found among experimental cheeses in terms of chemical composition or texture profile. Proteolytic activities were higher in cheeses kept in brine solutions that contained higher KCl (B, C, and D) compared with the control. At the end of the storage period, water-soluble nitrogen in Nabulsi cheeses stored in B, C, and D was higher than that in the control cheese (A). In addition, total viable count increased significantly after 1 mo of storage for all salt treatments. Hardness and gumminess generally decreased significantly during storage within the same salt treatment.     

Effect of chemico-physical parameters on the histidine decarboxylase (HdcA) enzymatic activity in Streptococcus thermophilus PRI60

In this study the activity of the histidine decarboxylase (HdcA) of Streptococcus thermophilus PRI60 was determined during growth and in crude enzyme preparations to evaluate its hazardousness in dairy products. The effect of different pH values, lactose availability, NaCl concentration, and growth temperature on histamine production was evaluated in M17 medium during 168 h incubation. In each case, the production of histamine increased concomitantly with the cell number with a relatively small further rise during the stationary phase. In all cultures the maximum histamine levels were reached at the end of active growth. Histamine was detectable (10 to 55 mg/L) even when growth was strongly inhibited. The HdcA enzyme in crude cell-free extracts was mostly active at acidic pH values common in dairy products. NaCl concentrations lower than 5% did not affect its activity. The enzyme was quite resistant to heat treatments resembling low pasteurization, but was inactivated at 75 °C for 2 min. Given the features of the enzyme studied, efforts must be dedicated to a thorough risk analysis and development of strategies to contrast the presence of histaminogenic S. thermophilus strains in products from raw or mildly heat-treated milk. PRACTICAL APPLICATION: During its growth Streptococcus thermophilus can produce histamine over a wide range of conditions encountered in cheesemaking and cheese ripening. The histidine-decarboxylase is even more active in cell-free extract and histamine can be accumulated independently of cell viability.     

The effect of substitution of NaCl with KCl on chemical composition and functional properties of low-moisture Mozzarella cheese

The effect of NaCl substitution with KCl on chemical composition, organic acids profile, soluble calcium, and functionality of low-moisture Mozzarella cheese (LMMC) was investigated. Functionality (meltability and browning), organic acids profile, and chemical composition were determined. Chemical composition showed no significant difference between experimental cheeses at same storage period, and same salt treatment. Meltability of LMMC salted with 3NaCl:1KCl, 1NaCl:1KCl, and 1NaCl:3KCl was higher compared with only NaCl (control). The amount of soluble Ca and P increased significantly during storage, with no significant difference between salt treatments. Organic acids profile did not differ between salt treatments at the same storage time.     

保加利亚乳杆菌与嗜热链球菌发酵夸克的工艺条件优化

以脱脂乳为原料,保加利亚乳杆菌和嗜热链球菌混合发酵剂制备夸克。以乳清OD500nm值、夸克校正产率及感官品质为评定指标,对凝乳酶添加量、CaCl2添加量、发酵剂嗜热链球菌与保加利亚乳杆菌配比、切割pH值4个因素进行考察。在此基础上对夸克感官评分进行四因素两水平的正交试验。结果表明：在凝乳酶添加量0.015%、嗜热链球菌与保加利亚乳杆菌配比3:2、CaCl2添加量0.02%、切割pH值为4.5时,所得夸克的感官评分最高,具有浓郁的奶香味,酸味清爽无颗粒感。     

Manufacture of low-sodium Minas fresh cheese: effect of the partial replacement of sodium chloride with potassium chloride

We investigated the effect of sodium reduction by partial substitution of sodium chloride (NaCl) with potassium chloride (KCl) on the manufacture of Minas fresh cheese during 21 d of refrigerated storage. Four treatments of low-sodium Minas fresh cheese were manufactured, with partial replacement of NaCl by KCl at 0, 25, 50, and 75% (wt/wt), respectively. The cheeses showed differences in the content of moisture, ash, protein, salt, and lipid contents, as well as on the extent of proteolysis and hardness throughout the storage period. However, no difference was observed among treatments within each storage day tested. The partial substitution of NaCl by KCl decreased up to 51.8% the sodium concentration of the cheeses produced. The consumer test indicated that it is possible to manufacture a low-sodium Minas fresh cheese that is acceptable to consumers by partial substitution of NaCl by KCl at 25% (wt/wt) in the salting step.     

Proteolytic profiles and angiotensin-I converting enzyme and alpha-glucosidase inhibitory activities of selected lactic acid bacteria

ABSTRACT:  This study was conducted to examine the growth, proteolytic profiles as well as angiotensin-I converting enzyme (ACE) and α-glucosidase (α-glu) inhibitory potentials of selected strains of lactic acid bacteria (LAB). Two strains each of yogurt bacteria ( Streptococcus thermophilus —1275 and 285, and Lactobacillus delbrueckii ssp. bulgaricus —1092 and 1368), and probiotics ( L. acidophilus —4461 and 33200, and L. casei —2607 and 15286, and 1 strain of Bifidobacterium longum 5022), were cultivated in reconstituted skim milk (RSM) at 37 °C and their proteolytic profiles and ACE as well as α-glu inhibitory activities were determined. Among all the strains of lactic acid bacteria studied, yogurt bacteria grew very well, with the exception of L. delbrueckii ssp. bulgaricus 1368 which showed a slower growth during the initial 3 h of incubation. The growth pattern corresponded well with the decrease in pH for the organisms. All the organisms showed an increase in proteolysis with time. The variations in proteolytic capabilities translated into corresponding variations in ACE inhibitory potential of these organisms. Bifidobacterium longum 5022 showed the highest ACE inhibitory potential followed by L. delbrueckii ssp. bulgaricus 1368, L. casei 15286, S. thermophilus 1275, and L. acidophilus 4461. Organisms with high intracellular enzymatic activities grew well. Also, aminopeptidases of strains of L. acidophilus 4461 and S. thermophilus 1275 that could better utilize proline containing substrates showed enhanced ACE inhibitory potential. Lactic acid bacteria possessed the ability to inhibit α-glu activity, which endowed them an antidiabetic property as well.     

The effect of NaCl substitution with KCl on Akawi cheese: Chemical composition, proteolysis, angiotensin-converting enzyme-inhibitory activity, probiotic survival, texture profile, and sensory properties

The effect of partial substitution of NaCl with KCl on Akawi cheese with probiotic bacteria was investigated during 30 d of storage at 4°C. Chemical composition, the survival of probiotic and lactic acid bacteria, proteolytic activity, and texture profile analysis were analyzed and sensory analysis was carried out to determine the effects of substitution. No significant differences were observed in moisture, protein, fat, and ash contents among the experimental Akawi cheeses at the same storage period. Significant differences were observed in water-soluble nitrogen and phosphotungstic-soluble nitrogen between experimental cheeses at the same of storage period. No significant difference was observed in the growth of Lactobacillus delbrueckii ssp. bulgaricus between experimental cheeses at the same storage period. However, the growth of Streptococcus thermophilus, Lactobacillus casei, and Lactobacillus acidophilus was significantly affected among experimental cheeses. A significant difference was observed in soluble Ca among experimental cheeses at the same storage period. In general, no significant differences existed in hardness and adhesiveness among experimental cheeses at the same storage period. No significant differences existed in sensory attributes, including creaminess, bitterness, saltiness, sour-acid, and vinegar taste among experimental Akawi cheeses at the same storage period.     

Effect of Versagel on the growth and metabolic activities of selected lactic acid bacteria

ABSTRACT:  In this study, the influence of a protein-based fat replacer, Versagel ^® added at 1% and 2% (w/v) to reconstituted skim milk (RSM), on the growth and metabolic activities of selected strains of yogurt starters ( Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus ) as well as probiotic organisms ( Lactobacillus casei , Lactobacillus acidophilus , and Bifidobacterium longum ) was examined. Addition of Versagel resulted in significantly improved growth of S. thermophilus and B. longum but inhibited that of L. casei , L. acidophilus , and L. delbrueckii ssp. bulgaricus . This is also reflected in the extent of reduction of pH in RSM with added Versagel by these organisms. Among the biochemical activities, proteolytic activity of all the organisms except B. longum was adversely affected by the presence of Versagel, although the angiotensin-I converting enzyme (ACE) inhibitory and α-glucosidase (α-glu) inhibitory activities were improved. Versagel at 1% level influenced the growth, while ACE inhibitory and α-glu inhibitory activities of the organisms were better at 2% level.     

Contribution of Streptococcus thermophilus to growth-stimulating effect of yogurt on rats

The origin of the growth-stimulating factor in yogurt was studied in rats fed liquid or freeze-dried diets of milk, yogurt, milks fermented individually by Streptococcus thermophilus and Lactobacillus bulgaricus, milks to which cells of Streptococcus thermophilus and Lactobacillus bulgaricus were added. Diets containing sonicated cells, cell supernatant, and cell fractions also were fed. Milk fermented by Streptococcus thermophilus and milk plus Streptococcus thermophilus cells stimulated growth as effectively as did yogurt. That finding and the absence of stimulation in rats fed Lactobacillus bulgaricus showed that Streptococcus thermophilus is responsible for stimulation of growth by yogurt. Growth was stimulated by an intracellular factor and not by fermentative changes in the milk.     

Salt Reduction in a Model High‐Salt Akawi Cheese: Effects on Bacterial Activity,pH, Moisture,Potential Bioactive Peptides,Amino Acids,and Growth of Human Colon Cells

This study evaluated the effects of sodium chloride reduction and its substitution with potassium chloride on Akawi cheese during storage for 30 d at 4 °C. Survival of probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium longum) and starter bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus), angiotensin‐converting enzyme‐inhibitory and antioxidant activities, and concentrations of standard amino acids as affected by storage in different brine solutions (10% NaCl, 7.5% NaCl, 7.5% NaCl+KCl [1:1], 5% NaCl, and 5% NaCl+KCl [1:1]) were investigated. Furthermore, viability of human colon cells and human colon cancer cells as affected by the extract showing improved peptide profiles, highest release of amino acids and antioxidant activity (that is, from cheese brined in 7.5% NaCl+KCl) was evaluated. Significant increase was observed in survival of probiotic bacteria in cheeses with low salt after 30 d. Calcium content decreased slightly during storage in all cheeses brined in various solutions. Further, no significant changes were observed in ACE‐inhibitory activity and antioxidant activity of cheeses during storage. Interestingly, concentrations of 4 essential amino acids (phenylalanine, tryptophan, valine, and leucine) increased significantly during storage in brine solutions containing 7.5% total salt. Low concentration of cheese extract (100 μg/mL) significantly improved the growth of normal human colon cells, and reduced the growth of human colon cancer cells. Overall, the study revealed that cheese extracts from reduced‐NaCl brine improved the growth of human colon cells, and the release of essential amino acids, but did not affect the activities of potential bioactive peptides.     

A method for determining beta-galactosidase activity of yogurt cultures in skim milk

A method was developed for determining the specific activity of bacterial beta-galactosidase (EC 3.2.1.23) during growth of Streptococcus thermophilus and Lactobacillus bulgaricus in skim milk. Individual and mixed strain cultures of S. thermophilus (St 3642, St14485) and L. bulgaricus (Lb11842, Lb880) were examined for growth (OD at 600 nm and viable cell counts), acid production, and beta-galactosidase activity (expressed as a function of recoverable TCA-precipitable cellular protein). Cultures were inoculated into 10% skim milk (2% inoculum) and incubated at 40 degrees C for 12 h. Aliquots were removed at 2-h intervals and diluted with ice cold EDTA, pH 12. The EDTA chelates calcium and solubilizes milk protein, allowing separation of the bacteria by centrifugation. Cells were then washed twice with 20 mM phosphate buffer and disrupted by sonication. Cell debris and intact cells were removed by centrifugation and the cell-free extract evaluated for beta-galactosidase activity using o-nitrophenyl-beta-D-galactopyranoside as substrate. Specific activities ranged from 0 to 6 units/mg protein. This simple and reproducible method is applicable for enzyme assays and measurement of cellular components where contamination by milk proteins is a potential problem.     

Angiotensin I converting enzyme-inhibitory activity of bovine, ovine, and caprine kappa-casein macropeptides and their tryptic hydrolysates

This work evaluated the angiotensin-converting enzyme (ACE)-inhibitory activities of bovine, ovine, and caprine kappa-casein macropeptides (CMPs) and their tryptic hydrolysates. The results obtained indicate that bovine, ovine, and caprine CMPs exhibited moderate in vitro ACE-inhibitory activities that increased considerably after digestion under simulated gastrointestinal conditions. Active peptides could also be produced from CMPs via proteolysis with trypsin, with tryptic hydrolysates exhibiting a more extensive ACE-inhibitory activity than intact CMPs during simulated gastrointestinal digestion. Two active fractions were chromatographically separated from the tryptic hydrolysate of the bovine CMP, but their complexity hampered the assignment of the ACE-inhibitory activity to specific peptide sequences. Evidence for the release of the strong ACE-inhibitory tripeptide IPP was found upon simulation of the gastrointestinal digestion of peptides released by trypsin from the CMP sequence. These findings might help to promote further exploitation of cheese whey in the preparation of nutraceuticals for inclusion in the composition of functional food products with high added values.     

Natural milk cultures for the production of Argentinian cheeses.

Samples (32) of natural milk cultures used in the Santa Fe, Argentina, area for soft and semihard cheese production were examined. The microbial composition (including lactic acid microflora characterization) and technological parameters (acidifying and proteolytic activities) were evaluated. The cultures contained mainly thermophilic lactic acid bacteria, identified as Streptococcus salivarius subsp. thermophilus (96.8% of the total strains) and Enterococcus spp. The strains showed a low proteolytic activity. The isolates of S. salivarius subsp. thermophilus exhibited a widespread phage resistance. The nonlactic microflora comprised coliforms, yeasts, spore-forming bacteria and lactate fermentative bacteria. The samples showed an acidity level from 0.38 to 0.69% lactic acid (pH from 4.25 to 5.75). The acidifying activity was optimal at 45 degrees C. The advantages and disadvantages of the employment of natural milk starters are discussed.     

钾盐替代和附属发酵剂对切达干酪品质的影响

为明确降低钠含量和添加在脱脂乳中表现出具有较高血管紧张素转换酶（angiotensin-converting enzyme,ACE）抑制率的Lactobacillus paracasei M3作为附属发酵剂对切达干酪理化性质和体外抗高血压潜力的影响,将部分钾盐替代和L. paracasei M3单独和混合加入到干酪与空白组作对照,以主要成分、微生物、蛋白水解度、质构和风味为理化指标和以ACE抑制率为体外抗高血压潜力指标。结果表明,4 组干酪的主要成分差异不大（P＞0.05）,钾盐替代组、L. paracasei M3组、钾盐替代和L. paracasei M3混合组干酪在乳酸菌总数、pH 4.6可溶性氮、总游离氨基酸上显著高于空白组干酪（P＜0.05）,但是降低了干酪的硬度、pH值和有明显的苦味（P＜0.05）,而在体外抗高血压潜力方面,3 组干酪在成熟6 个月之后ACE抑制率达65.2%、74.3%和78.7%,比空白组分别高19.6%、36.2%和44.4%,由此可知,在切达干酪中钾盐替代和添加L. paracasei M3有助于具有抗高血压潜力的ACE抑制肽的产生,但是后续需要对风味进行补偿性研究。     

Effects of somatic cell count and stage of lactation on the plasmin activity and cheese-making properties of ewe milk

The experiment was conducted from March to July 2002 using 5 intensively managed flocks of Southern Italy. In each flock, 2 groups of 50 ewes were created. The groups were designated LSCC (low somatic cell count [SCC]) when their milk SCC was lower than 500,000/mL and HSCC (high SCC) when their milk SCC was higher than 1,000,000/mL. Bulk milk and whey samples were analyzed for fat, total protein, lactose, casein, and whey protein contents. Renneting properties of milk were also determined. Moisture, NaCl, and nitrogen fractions were determined in fresh cheese curds. In addition, plasmin (PL) and plasminogen (PG) activities in milk and cheese were monitored. The proteolytic activity of plasmin by urea-polyacrylamide gel electrophoresis and the white blood cell (WBC) differentials were determined. The HSCC resulted in higher pH values in milk and in higher moisture and lower fat contents in fresh cheese curds. Moreover, a lower recovery of fat and whey proteins was obtained from the HSCC than from the LSCC raw milk. The crude protein and casein contents were higher in the HSCC than in the LSCC curds during early and midlactation; an opposite trend was observed in late lactation. Plasmin and PG activities underwent more marked fluctuations in the LSCC than in the HSCC curds through lactation. The results of this experiment demonstrate that the PL activity in ewe milk is markedly influenced by the SCC, although SCC is not the only parameter for predicting PL and PG evolution in ewe milk. The LSCC milk resulted in a higher proteolytic potential of Canestrato pugliese cheese curds.     

Functional Properties of Protein Fractions Obtained from Pumpkin (Cucurbita Maxima) Seed

Protein fractions were extracted from pumpkin (Cucurbita maxima) seeds with deionized water, salt solution, and alkali solution, and characterized under alkaline (pH 8) and ionic strength conditions for protein solubility, interfacial pressure, and functional properties. The supernatants obtained from the protein fractions after hydration and centrifugation were designed deionized water supernatant, salt solution supernatant, and alkali solution supernatant. All protein fractions showed poor foamability and very low stability. The designed deionized water supernatant presented the highest emulsifying properties. In terms of stability, designed deionized water supernatant emulsions flocculate during storage but with low rates of coalescence compared to the two other fractions.