Crystallization properties and polymorphism of triacylglycerols in goat's milk fat globules

The sensorial, functional, and nutritional properties of goat dairy products result from the specific fatty acid composition of goat's milk fat. However, information on the physical and thermal properties of goat's milk fat is scarce. In this study, crystallization of triacylglycerols (TG) in goat's milk fat globules was investigated using polarized light microscopy and the coupling of time-resolved synchrotron radiation X-ray diffraction (XRD) and high-sensitivity differential scanning calorimetry (DSC). The molecular organization of the solid fat phase was characterized for cooling rates between 3 and 0.1 degrees C/min. Quenching of goat's milk fat globules from 50 to -8 degrees C and 4 degrees C was also examined to identify the most unstable polymorphic forms of TG. Then, the melting behavior of fat crystals was studied on subsequent heating at 1 degrees C/min. Triple chain length (3L: 68.6-70 A) and double chain length (2L: 37-45.4 A) structures were characterized and 5 polymorphic forms, alpha, sub-alpha, beta' 1, beta' 2, and beta were identified. Polymorphic transitions were observed within goat's milk fat globules as a function of time after quenching and as a function of temperature on heating. From a technological point of view, this work will contribute to a better understanding of the rheological properties as well as on the flavor evolutions of goat's milk-based products.     

Transmission electron microscopy of mozzarella cheeses made from microfluidized milk.

The nanostructure of Mozzarella cheeses prepared from microfluidized milk was compared with that of control cheeses made from untreated milk. Milk heated to 10 or 54 degrees C and containing 1.0 or 3.2% fat was homogenized by microfluidization at 34 or 172 MPa prior to cheesemaking. The effects on the casein particles and fat globules in the cheese were determined by transmission electron microscopy after 1 day and 6 weeks of storage at 4 degrees C. The micrographs showed that electron-dense regions theorized to be casein submicelles rearranged from a homogeneous configuration to a pattern of clusters during the storage period. The nanostructure of the cheeses made from milk processed under the mildest conditions resembled the controls, but otherwise the fat droplets decreased in size and increased in number as the pressure and temperature were increased. The results indicate that both homogenization temperature and pressure affect the nanostructure of Mozzarella cheese.     

Multiscale characterization of the organization of triglycerides and phospholipids in emmental cheese: from the microscopic to the molecular level

The chemical composition and properties of lipids, both triglycerides and phospholipids, play a major role in the functional and nutritional properties of food products. In this study, the suprastructure of fat, solid fat content, and crystallographic properties of triglycerides were investigated in hard-type cheeses from the microscopic scale to the molecular level using the combination of relevant techniques. Two industrial cheeses with different oiling off properties were compared with experimental cheeses manufactured in the laboratory. Microstructural analysis performed using confocal laser scanning microscopy showed that milk processing led to the disruption of fat globules with the formation of nonglobular fat. For a similar fatty acid composition, oiling off was mainly related to the fat in dry matter content and to the suprastructure of fat in cheese. An exogenous fluorescent phospholipid permitted the localization of milk phospholipids in the cheese matrix, which mainly remain around fat inclusions after disruption of the milk fat globule membrane, and to show heterogeneities. We also showed using differential scanning calorimetry that the suprastructure of fat did not affect the solid fat content in cheese at 4 degrees C: 71.6 +/- 4.9%. The organization of triglyceride molecules in fat crystals, elucidated at a molecular level using X-ray diffraction, corresponded to the coexistence of 2 lamellar structures (2L 40.5 angstroms and 3L 54.6 angstroms) with four polymorphic forms: alpha, two beta' and beta. A schematic representation of the multiscale organization of triglycerides and phospholipids in cheese is proposed.     

Chemical and microbiological characteristics of ewes' milk cheese manufactured with extracts from flowers of Cynara cardunculus and Cynara humilis as coagulants

The chemical and microbial characteristics as well as the flavor and aroma of Los Pedroches cheese made using aqueous extracts of Cynara cardunculus L. flowers were compared with those of cheeses manufactured with extracts of Cynara humilis L. throughout ripening. The two thistle species assayed were found to have no appreciable effect on the moisture, fat, protein, and NaCl contents of the cheese or on its water activity, flavor, and aroma; however, the use of C. humilis resulted in reduced lactic acid content (p < 0.001) and higher pH values (p < 0.05) relative to those of cheese specimens produced with C. cardunculus. The protein breakdown of the cheeses was assessed in terms of soluble nitrogen (SN), nonprotein nitrogen (NPN), and amino acid nitrogen (AAN). Proteolysis was more marked and rapid in cheese containing C. cardunculus as coagulant, the SN and NPN contents of which were significantly higher (p < 0. 01) than those of the cheese obtained with the species C. humilis; AAN contents were similar in both species of Cynara throughout ripening. Although total viable, coliform, and lactobacilli counts were similar in cheeses produced with both types of plant coagulant throughout ripening, enterobacteria and yeasts counts (p < 0.01) and molds counts (p < 0.05) were higher in cheese produced with C. humilis than in cheese obtained with C. cardunculus.     

Rheological and proteolytic properties of Monterey Jack goat's milk cheese during aging

To enhance the understanding of the quality traits of goat's milk cheeses, rheological and proteolytic properties of Monterey Jack goat's milk cheese were evaluated during 26 weeks of 4 degrees C storage. As expected with aging, beta-casein levels decreased with concomitant increases in peptide levels and were correlated with changes in rheological properties of the cheese. Hydrolysis of the protein matrix resulted in more flexible (increased viscoelastic properties) and softer (decreased hardness, shear stress, and shear rigidity) cheeses. During the first 4-8 weeks of storage, cheese texture changed significantly (P < 0.05) and then stabilized. Characterization of rheological and proteolytic properties of the goat's milk semihard cheese during aging provided insight into the changes occurring in the protein matrix, the relationship to structure, and a shift in cheese quality.     

Flavor enhancement of reduced fat cheddar cheese using an integrated culturing system

Mild cheese flavor in reduced fat Cheddar cheese was enhanced by using an integrated starter culture system. Three cultures, Lactococcus lactis subsp. cremoris SK11, L. lactis subsp. lactis biovar. diacetylactis JVI, and Lactobacillus casei 7A, were carefully selected to obtain a nonbitter, mildly acid, buttery flavored cheese. Cheeses were produced from all possible combinations of these cultures with the constraint that L. lactis subsp. cremoris SK11 was used as the primary acid-producing culture. Cheeses made with SK11 were compared to cheeses produced using an L. lactis subsp. cremoris commercial starter culture. Cheeses were ripened for 150 days and periodically sampled for chemical, microbiological, and sensory analysis. Cheeses produced with L. lactis subsp. cremorisSK11 had substantially lower bitterness intensity than the cheeses produced with commercial starter culture. L. lactis subsp. lactis biovar. diacetylactis JVI significantly increased diacetylacetoin and acetate concentrations. Sensory results indicate that these cheeses had increased buttery (diacetyl) flavor.     

Ethyl ester formation is enhanced by ethanol addition in mini Swiss cheese with and without added propionibacteria

Esters are important contributors to cheese flavor, but their mechanisms of synthesis in cheese are largely unknown. This study aimed to determine whether ethanol concentration limits the formation of ethyl esters in cheese. Mini Swiss cheeses were manufactured with (E) or without (C) the addition of ethanol to cheese milk. Ethanol concentrations (enzymatic analysis) were 64 +/- 17 and 330 +/- 82 microg g(-1), respectively, in C and E cheeses. E cheeses also contained 5.4 +/- 2.3 times more of the five ethyl esters quantified than C cheeses, regardless of the concentrations of esters in C cheeses (range 1-128 ng g(-1)). Furthermore, the presence of propionibacteria added as acid-producing secondary starters was associated with greater concentrations of esters, due to the increase in acid concentrations that propionibacteria induced and/or to an involvement of propionibacteria enzymes in ester synthesis. This study demonstrates that ethanol is the limiting factor of ethyl ester synthesis in Swiss cheese.     

Wavelength dependence of light-induced lipid oxidation and naturally occurring photosensitizers in cheese

Degradation of the potential photosensitizers, riboflavin, chlorophyll, and porphyrin, in Danbo cheese by monochromatic light of wavelength 366, 436, or 546 nm was studied. Three cheeses were investigated, two conventional (16% fat and 25% fat) and one "organic" (25% fat). The effect of illumination was measured by fluorescence spectroscopy and analyzed using multiway and multivariate data analysis. Riboflavin was found to degrade only by 436 nm light, whereas chlorophylls and porphyrins also were influenced by 436 and 546 nm light. The organic cheese had the largest chlorophyll content both before and after similar light exposure, and no change in chlorophyll of this cheese was observed for any of the illumination wavelengths. Upon light exposure of the cheeses, volatile compounds were formed, as analyzed by gas chromatography-mass spectrometry (GC-MS). The relative concentrations of methyl butanoate, 1-pentanol, benzaldehyde, 2-butanone, 2-heptanone, and butyl acetate were found to weakly correlate with the surface fluorescence intensity. 1-Pentanol and the ketones are secondary lipid oxidation products, consistent with a chemical coupling between photosensitizer degradation and formation of volatile lipid oxidation products.     

Time course and specificity of lipolysis in Swiss cheese

Controlling lipolysis in cheese is necessary to ensure the formation of desirable flavor. To get a better understanding of the mechanism of lipolysis in Swiss cheese, cheeses were manufactured with and without (control) the addition of Propionibacterium freudenreichii. Products of lipolysis were quantified throughout ripening. Half of the free fatty acids (FFA) released in milk (3.66 mg/g fat), in particular the short-chain FFA, were lost in the whey during curd drainage, whereas diglycerides and monoglycerides were retained within the curd. P. freudenreichii was responsible for the release of most FFA during ripening (10.84 and 0.39 mg/g fat in propionibacteria-containing and control cheeses, respectively). Indices of lipolysis displayed low specificity. All types of FFA were released, but butyric and palmitic acids more significantly, which could be due to a low sn-1,3 regioselectivity. All glycerides were hydrolyzed in the following order: monoglycerides>diglycerides>triglycerides. The results of this study show the quantitative and qualitative contributions of the different lipolytic agents to Swiss cheese lipolysis.     

Vitamin D3 fortification, quantification, and long-term stability in Cheddar and low-fat cheeses

Considering the widespread insufficiency of vitamin D, the fortification of additional foods with vitamin D is warranted. The objective of this research was to assess the feasibility of vitamin D3 fortification in natural hard cheeses. We examined the recovery, distribution, long-term retention, and heat stability of the vitamin in industrially made fortified Cheddar and low-fat cheeses. The results indicated that the vitamin D3 did not degrade during processing, over 1 year of ripening (3-8 degrees C), or after thermal treatment at 232 degrees C for 5 min. Vitamin D3 recovery in the fortified Cheddar and low-fat cheeses were, respectively, 91 and 55% of the vitamin D3 added to the milk used to make each cheese. The remaining vitamin D3 was entrained in the whey. The vitamin D3 was uniformly distributed throughout the blocks of cheese. The fortification process did not alter the yield, chemical composition, or flavor of the Cheddar cheese. We conclude that industrially manufactured Cheddar and low-fat cheeses are suitable for vitamin D3 fortification.     

Conjugated linoleic acid in processed cheeses during the manufacturing stages

Conjugated linoleic acid (CLA) is a naturally occurring micronutrient in milk fat and dairy products consisting of a group of geometric and positional isomers. The purpose of this study was to assess the level and type of CLA isomers found in two commercial processed cheeses (portions and slices) as well as to monitor their evolution during the different manufacturing stages. Total CLA concentrations ranged from 7.5 to 7.9 mg/g of fat, and rumenic acid (cis-9,trans-11 C18:2), the isomer responsible for the biological functions, represented >80% of total CLA. trans-11,cis-13 and trans-11,trans-13 were, with approximately 4% each, the second main CLA isomers. trans-trans isomers accounted for <10% of total CLA. The processing parameters used in this research had negligible effects on the CLA content of processed cheese and did not modify the isomer profile in these dairy products, thereby confirming the stability of rumenic acid during manufacturing.     

产脂肪酶乳酸菌对新疆传统奶酪脂肪酸及风味的影响

为了改善奶酪品质,奶酪生产过程中通常会添加脂肪酶或者产脂肪酶乳酸菌来提升产品品质。该研究以前期筛选的4株高产脂肪酶乳酸菌为发酵剂,分别随机选取3株乳酸菌复配制作酸凝奶酪。试验组：A组T1-5和T1-3属融合魏斯氏菌（Weissella confusa）、H1-6属瑞士乳杆菌（Lactobacillus helveticus）,B组H1-6、T1-5、B2-5属植物乳杆菌（Lactobacillus plantarum）,C组H1-6、T1-3、B2-5,D组T1-3、T1-5、B2-5,对照组（E组）（添加商业发酵剂）,分析发酵剂对传统奶酪pH值、滴定酸度和脂肪氧化情况的影响,并利用气相色谱法（Gas Chromatography,GC）检测奶酪中脂肪酸变化、利用气相色谱-离子迁移谱（Gas Chromatography-Ion Mobility Chromatography,GC-IMS）分析奶酪中风味物质的变化。结果表明：A,B,C,D组4组奶酪的pH值、过氧化值（Peroxide value,POV值）明显低于E组（对照组）（P < 0.05）,A,B组奶酪滴定酸度比对照E组高（P < 0.05）;A,B,C,D组奶酪中饱和脂肪酸（Saturated Fatty Acids,SFA）含量、单不饱和脂肪酸（Monounsaturated Fatty Acids,MUFA）含量和多不饱和脂肪酸（Polyunsaturated Fatty Acids,PUFA）含量均显著高于E组（P < 0.05）;4个试验组样品中亚油酸（C18∶2n6c）含量明显高于对照组（E组）（P < 0.05）。GC-IMS及主成分分析结果显示,A、B组奶酪挥发性风味物质种类多,且相似度较高,其中2-庚酮、丁醛、乙酸丁酯是主要呈味物质;C、E两组奶酪中风味物质比较相似,风味物质主要以乙酸乙酯、乙酸丙酯、己酸乙酯等酯类为主;D组与其他4组有所差异,主要挥发性风味物质为乙酸丁酯、3-辛酮、庚醛等。结合感官评定,A、B两组奶酪整体风味和口感较好,评分较高。筛选得到的产脂肪酶乳酸菌可以作为发酵剂用于提升新疆传统奶酪品质。     

The addition of Propionibacterium freudenreichii to Raclette cheese induces biochemical changes and enhances flavor development

Two mixtures of Propionibacterium freudenreichii commercial strains were tested as adjunct cultures in pasteurized milk Raclette cheese to investigate the ability of propionibacteria (PAB) to enhance flavor development. Cheese flavor was assessed by a trained sensory panel, and levels of free amino acids, free fatty acids, and volatile compounds were determined. The PAB level showed a 1.4 log increase within the ripening period (12 weeks at 11 degrees C). Eye formation, which was not desired, was not observed in PAB cheeses. PAB fermented lactate to acetate and propionate and produced fatty acids by lipolysis, branched chain volatile compounds derived from isoleucine and leucine catabolism and some esters. One of the experimental cheeses received the highest scores for odor and flavor intensity and was characterized by higher frequencies of detection for some minor notes ("propionic"and "whey" odor, "sweet" taste). PAB can therefore be considered as potential adjunct cultures to enhance or modify cheese flavor development.     

Identification and enumeration of beta-hemolytic Listeria monocytogenes in naturally contaminated dairy products

A DNA probe was used to identify hemolytic Listeria monocytogenes in naturally contaminated dairy products: unpasteurized milk, ricotta cheese, and imported semisoft cheeses. Of 34 milk samples, 12 were suspected to contain hemolytic L. monocytogenes; 1 contained greater than 6000 viable organisms/g. The ricotta cheese, although temperature-abused, had a titer of 3.6 x 10(6) beta-hemolytic L. monocytogenes cells/g, whereas the semisoft cheeses reached a maximum of 5.6 x 10(6) cells/g. Pure cultures of L. monocytogenes isolated from both types of cheese were found positive by the CAMP test and the DNA probe.     

Effect of variety and maturation of cheese on supercritical fluid extraction efficiency

Supercritical fluid extraction (SFE) has been utilized by the food industry in many applications to extract, fractionate, and recover compounds from various food matrices. However, little research has been conducted using SFE as an alternative process for producing reduced-fat cheese. Lipids in cheeses may be selectively extracted due to the nonpolar properties of supercritical carbon dioxide (SC-CO2), without leaving residual chemicals as is the case in solvent extraction. The objective of this study was to evaluate the influence on the extraction process due to cheese variety and protein breakdown by age. A Latin square design was utilized to test the extractability of lipids from Parmesan and Cheddar cheeses, aged young (9-10 months) or old (24 months). Extraction took place in a 500 mL SFE vessel using 100 g of grated cheese samples. The SFE parameters of the extraction were 350 bar, 35 degrees C, and supercritical carbon dioxide at a flow rate of 20 g/min for 55 min. Compositional analysis measured all treated samples and controls of total lipids, lipid profiling, total protein, protein/peptide analysis, moisture, ash, and pH. Cheese type was a major variable in fat extraction. The extraction in Cheddar showed an average fat reduction of 53.56% for young cheese, whereas that in old Cheddar was 47.90%. However, young Parmesan was reduced an average of 55.07%, but old Parmesan was reduced at 68.11%, measured on a dry basis. SFE extracted triglycerides and cholesterol, but did not remove phospholipids. This investigation introduces the observations of the effect of Cheddar and Parmesan varieties on SFE, offering data on the important parameters to consider in the design of SFE processes to reduce fat in cheese.     

Evaluation of APHA and AOAC methods for phosphatase in cheese

Varieties of market cheese were analyzed for alkaline phosphatase by the modified rapid colorimetric method of the American Public Health Association (APHA) and the official AOAC method, 16.304-16.306. In the APHA method, 5 g cheese (pH less than 7.0) is macerated with 2 mL 1:1 carbonate buffer, or 2 mL water (for cheese with pH greater than 7.0). Addition of 0.1 mL magnesium acetate (1 mg magnesium) to test portions of cheese extracts yielded reproducible and quantitative recovery of added phosphatase. In the AOAC method, macerating 0.5 g cheese with 1 mL borate buffer before adding milk phosphatase improved recovery among cheeses. Addition of magnesium ion increased phosphatase activity in some cheeses. Phosphatases in blue mold-ripened and Swiss cheeses were inactivated by heat faster than was milk phosphatase, yet milk phosphatase added to various soft cheeses was completely inactivated at 60 degrees C for 10 min. The lability of phosphatase was due to the heat-denaturing effect of NaCl present in finished cheeses. Some Mexican style soft cheeses contained both heat-labile and heat-stable phosphatases. These data suggest that the phosphatase test to differentiate milk and microbial phosphatases on the basis of repasteurization and analysis of cheese is no longer valid.     

Impact of whey pH at drainage on the physicochemical, sensory, and functional properties of mozzarella cheese made from buffalo milk

In this study, the effects of whey pH at drainage on the physicochemical, sensory, and functional properties of mozzarella cheese made from buffalo milk during storage were investigated. Four cheese samples were manufactured using starter culture at different whey pH values [(A) 6.2, (B) 5.9, (C) 5.6, and (D) 5.3] and analyzed on the 1st, 28th, and 56th day. Ash, calcium, and phosphorus concentrations decreased as the whey pH at drainage was lowered. Cheese yield and calcium recovery were the highest in D cheeses. During storage, expressible serum levels decreased and nonexpressible serum levels increased, indicating an increase in the water holding capacity of the cheeses. Reducing the calcium content of cheeses increased meltability values, but an overly low calcium level (D cheeses) had an adverse effect on the meltability. The melting properties of cheese samples, except D cheeses, were improved with aging. A cheeses were the hardest and D cheeses the softest throughout storage. The 1st day sensory evaluations revealed that C and D cheeses were preferred and that A cheeses were not. All sensory properties of A cheeses were improved with storage. D cheeses were rated inferior to the others at the end of the storage time.     

Effect of cooling rate on nucleation behavior of milk fat--sunflower oil blends.

The effect of cooling rate on the crystallization behavior of mixes of high-melting milk fat fraction (HMF) and sunflower oil (SFO) was studied by following nucleation with laser-polarized turbidimetry. The initial crystals were photographed, and their thermal and polymorphic behaviors, as well as chemical composition, were investigated by calorimetry, X-ray diffraction, and capillary gas chromatography. Activation energies of nucleation were calculated using the Fisher-Turnbull equation. Despite small differences in Mettler dropping points for different ratios of SFO to HMF, induction times were significantly different between samples and were shorter at a slow cooling rate for the same supercooling. Rapidly cooled samples required more time at crystallization temperature to crystallize than slowly cooled samples because molecular organization prior to nuclei formation took place under different conditions. Regardless of cooling rate or composition, all crystals were in the beta' polymorph. However, morphology, thermal behavior, and chemical composition showed differences with cooling rate. Activation free energies of nucleation were of the same order of magnitude as those published for hydrogenated SFO.     

Physicochemical variables affecting the rheology and microstructure of rennet casein gels

The rheology and microstructure of a rennet casein system were studied in the pH range from 5.8 to 12.0 during cooling from 80 to 5 degrees C at four cooling rates: 0.5, 0.1, 0.05, and 0.025 degrees C/min. A dramatic increase in storage modulus with pH was observed during cooling at a fixed cooling rate. Continuous networks were formed for gels at pH 7.2 and above, while a discontinuous network was observed for gels below pH 6.5. The monotonic increase in storage modulus with pH could be correlated to the number of net (negative) charges and the strength of the hydrophobic interactions. At a higher pH, the protein micelles were larger due to weaker hydrophobic interactions and stronger repulsive electrostatic interactions resulting from more charges. When these protein micelles aggregated into flocs during cooling, the flocs had similar sizes at different pH values but a smaller fractal dimension at a higher pH. Consequently, for systems of the same protein and salt concentrations, more flocs were present in the gels at a higher pH, which subsequently generated more cross-links and a higher storage modulus. The pH also determined how the cooling rate affected the gel properties. At pH 5.8 and 6.5, the gels were firmest at the fastest cooling schedule, and the cooling rate did not show a trend in affecting the gel strength at the other three rates. On the other hand, a slower cooling rate generated a firmer gel at pH 7.2 and 12.0. The analysis of casein interactions suggests that the cooling rate affected the casein floc size only when repulsive interactions enabled a slow flocculation (at higher pH values) comparable with temperature change rates during cooling. For rennet casein gels of pH within the range of processed cheese products (pH 5.8 and 6.5), particle or cluster rearrangements created more uniform networks for gels cooled at slower schedules and weakened the structure.     

Functional properties of oat globulin modified by a calcium-independent microbial transglutaminase

Oat globulin was modified by a calcium-independent microbial transglutaminase (TG). The TG-polymerized protein had higher solubility than the control at acidic pH and had improved water- and fat-binding properties. Incubation of 10% (w/v) oat globulin dispersions in the presence of TG at 37 degrees C led to the formation of a well-developed viscoelastic gel network with a microstructure characterized by thick strands and large clusters. The TG-induced gels had higher modulus values, lower loss tangent values, and lower frequency dependency than the heat-induced gels. The TG-induced gel system has the characteristics of classical polymer gel with permanent "chemical" cross-links, whereas the heat-denatured system has the characteristics of a temporary "physical" gel with breakable cross-links. Fourier transform infrared spectroscopy showed marked shift and intensity changes in several major bands, suggesting pronounced changes in protein conformation during TG-induced gelation. Aggregation of protein molecules was also indicated by the progressive increases in two infrared bands (1679-1682 and 1622-1625 cm(-)(1)) associated with the formation of intermolecular beta-sheets and strands. Results suggest that new food polymers with unique functionality can be produced from oat globulin treated with TG and that elastic gels can be formed near neutral pH, instead of the alkaline pH required for thermally induced oat globulin gels.