Triacylglycerol composition of protected designation of origin cheeses during ripening. Authenticity of milk fat

Triacylglycerol (TAG) composition by carbon number in 2 protected designation of origin cheeses, Mahón (cheese from cow milk) and Manchego (cheese from ewe milk) that were manufactured by 3 different producers was analyzed during cheese ripening using gas chromatography with a short capillary column. The TAG composition at different times during cheese ripening was also analyzed in cheeses from different batches produced at the same plant. Lipolysis levels in the Mahón and Manchego cheeses during ripening were low; free fatty acid values ranged from 2,500 to 4,000 ppm at the end of ripening. The TAG composition did not change significantly during ripening. The TAG values obtained from each cheese sample were substituted into the multiple regression equations that have been proposed to detect foreign fats in milk fat. The values obtained using the equations for bovine (proposed by the European Union) and ovine milk (proposed by our laboratory) were within the normal range. Accordingly, these equations can be considered useful for detecting foreign fat in these cheeses during the ripening period contemplated during this study.     

Effect of artisanal kid rennet paste on lipolysis in semi-hard goat cheese

This study examined the use of hygienised kid rennet pastes in model cheese systems and also in goat milk semi-hard cheeses to promote lipolysis. The results obtained indicated that the use of rennet paste caused greater lipolysis and increased, mostly, the short-chain free fatty acid (FFA) content. The model systems made with whole goat’s milk using rennet paste and commercial rennet mixture exhibited a higher FFA content than did the rennet paste-free controls (31,600 vs. 25,600 μmol/kg cheese). For the pilot cheeses made with bovine rennet and rennet paste mixture, the increase in FFA level after 45 days of ripening compared with the cheeses prepared only with commercial rennet was as much as 6600 (μmol/kg cheese) and the increase in the butyric acid content was also 1650 (μmol/kg cheese). Moreover, after 15 days of ripening, industrially prepared cheeses made with rennet paste exhibited greater levels of FFA than did the cheeses made with commercial rennet (11,500 μmol/kg at 45 days of ripening). Their flavour was stronger and the organoleptic characteristics were better accepted, which implies less ripening time for commercial cheese manufacture.     

Identity of the major triacylglycerols in ovine milk fat

Ovine milk fat obtained from milk collected from five different flocks was studied to determine its triacylglycerol (TAG) composition using a combination of gas chromatography/mass spectrometry. One hundred and thirty-four molecular species of TAG were identified, but not all could be quantified because some peaks contained more than one species of TAG. Fifty-one per cent of the identified species were trisaturated TAGs, 31% as monounsaturated TAGs and 18% as polyunsaturated TAGs. In terms of chain length, 58 of the TAGs identified (43% of the total) contained short-chain (C4 and C6) fatty acids (FAs), 94 (70%) contained medium-chain FAs (C8–C14) and 112 (84%) were composed of long-chain FAs (C16–C18). One hundred and twenty nine chromatographic peaks were detected. The sum of the 35 peaks, each representing>1 mol% of the TAGs, was 75% of total TAGs. Five peaks, each with>3.5 mol% included trisaturated and monounsaturated TAGs; quantitatively, the largest of these peaks (4.5 mol% of the total TAGs) contained two esterified monounsaturated TAGs with C4: 4,16,18:1 or 4,18,16:1, with the former being present at the highest levels. Comparison of the experimental values of TAG composition in ovine milk with the theoretical values derived from the experimental FA content showed that the distribution of the FAs in the TAGs was not random. The TAGs containing a short-chain FA were synthesized preferentially to those TAGs containing three medium- or three long-chain FAs. Our results for ewes’ milk were very similar to those for cows’ and goats’ milk despite the quantitative differences in the TAG profiles of the different species.     

Changes of composition and free fatty acid contents of Urfa cheeses (a white-brined Turkish cheese) during ripening: Effects of heat treatments and starter cultures

The influences of heat treatments (at 65 °C for 20 min or 72 °C for 5 min) applied to the milk and addition of mesophilic or thermophilic starter cultures, prior to cheese-making, on the composition and free fatty acid contents of Urfa cheeses were evaluated throughout the ripening period. Sensory evaluation of cheese samples was also performed on 90th day. The basic composition of ripened cheese samples was not significantly affected by the heat treatments and starter cultures. Heat treatments adversely affected the lipolysis and sensory properties of Urfa cheeses, particularly at 72 °C. The FFA contents of cheeses made from mesophilic and thermophilic cultures were similar. Cheese made from raw milk had a higher level of lipolysis than the cheeses made from milk inoculated with mesophilic or thermophilic lactic starters (p < 0.05).     

Effects of high-pressure treatment on free fatty acids release during ripening of ewes' milk cheese

The free fatty acid (FFA) profile of high pressure treated ewes' milk cheeses were studied to assess the effect of pressure treatment on cheese lipolysis. Cheeses were treated at 200, 300, 400 or 500 MPa (2P to 5P) at two stages of ripening (after 1 and 15 days of manufacturing; P1 and P15) and FFA were assayed at 1, 15 and 60 d ripening. On the first day of ripening, 3P1-cheeses showed levels of FFA twice that of the control cheeses. However, no significant differences were found between 3P1 and control cheeses at 60 d ripening. On the contrary, 4P1 and 5P1-cheeses had the lowest total FFA levels. The point at which pressure treatment was applied influenced the FFA profile of cheeses; cheeses pressurized at pressures<400 MPa on the first day of ripening were more similar to untreated cheeses than their homologues treated at 15 d.     

Effect of a bacteriocin-producing Lactococcus lactis strain and high-pressure treatment on the esterase activity and free fatty acids in Hispánico cheese

The effect of milk inoculation with a bacteriocin-producing (BP) culture and of high-pressure (HP) treatment of 15-day-old Hispánico cheeses (400 MPa, 5 min, 10 °C), separately or combined, on the release of intracellular esterases and cheese lipolysis was investigated. Esterase activity and free fatty acids (FFAs) content increased during ripening of Hispánico cheese and palmitic, oleic and stearic acids being the most abundant FFAs. On day 15, the highest esterase activity was recorded for HP-treated BP cheese. The activity for HP-untreated BP cheese was the next highest. No difference in the activities was found between HP-treated and untreated cheeses made without BP culture. Total FFAs on day 15 were at a lower concentration in BP cheeses than in cheeses made without BP culture, probably due to the lower pH values of the former. The rate of total FFA accumulation from day 15 to day 50 was higher in BP cheeses (31.1–32.1% increase) than in cheeses made without BP culture (19.3–21.7% increase). The highest total FFA concentration on day 50 (612 mg kg⁻¹) was found for HP-untreated cheese made without BP culture.     

Effect of high-pressure treatments on proteolysis and texture of ewes’ raw milk La Serena cheese

La Serena cheeses, made from Merino ewes’ raw milk, were high-pressure (HP)-treated at 300 or 400 MPa for 10 min at 10 °C, on days 2 or 50 of ripening. Cheeses treated by HP on day 2 showed higher pH values than control cheese on day 3, but cheeses treated by HP on days 2 or 50 and control cheese had similar pH values on day 60. Breakdown of caseins was delayed by HP treatment of cheeses on day 2. Cheeses treated by HP on day 2 showed higher levels of hydrophilic peptides, lower levels of hydrophobic peptides, lower hydrophobic peptides: hydrophilic peptides ratios, and higher total contents of free amino acids than those of control cheese. HP treatment of cheese on day 50 scarcely affected proteolysis of 60-day-old cheeses. Fracturability, hardness and elasticity values of cheeses treated by HP on day 2 were higher than those of control cheese and of cheeses treated on day 50. Cheeses treated at 400 MPa on day 2 received the lowest scores for quality of taste from panellists, whereas the rest of HP-treated cheeses did not differ from control cheese.     

Lipolysis in Urfa cheese produced from raw and pasteurized goats’ and cows’ milk with mesophilic or thermophilic cultures during ripening

Lipolysis was evaluated in Urfa cheese made from raw and pasteurized goats’ and cows’ milk with mesophilic or thermophilic cultures. The acid degree values (ADVs) of the cows’ milk cheeses were significantly (P < 0.05) higher until 60 d of storage than that of cheese made from goats’ milk. Total free fatty acid (FFA) contents of goats’ milk cheese were significantly (P < 0.001) lower than that of cows’ milk cheese throughout ripening, whereas goats’ milk cheese flavour was higher (P < 0.05) than cows’ milk cheese. Pasteurization of milk prior to cheese-making has a negative influence, not only on the level of lipolysis throughout ripening, but also on the relative amounts of short chain FFAs and sensory properties of the cheeses (P < 0.001). Cheese produced without starter bacteria underwent significantly (P < 0.05) higher lipolysis than cheeses produced with mesophilic or thermophilic starter bacteria, while cheese made with thermophilic starter culture had similar flavour to cheese made without starter culture.     

Free fatty acids and oxidative changes of a raw goat milk cheese through maturation

Free fatty acids (FFA) and lipid and protein oxidation changes were studied throughout maturation process of a raw goat milk cheese with protected designation of origin. Cheeses were analyzed at 4 different times of maturation, at 1, 30, 60, and 90 d. All FFA significantly increased during maturation and the relative increase was higher for long-chain than medium- or short-chain FFA. At the end of maturation, oleic (C18:1 n9), butyric (C4:0), and palmitic (C16:0) acids were the most abundant. The higher levels of short-chain fatty acids (SCFA) regarding total FFA obtained at the end of Ibores cheese ripening compared with other raw goat milk cheeses, highlight the notable role of SCFA on the flavor of this cheese owing to their low-odor thresholds. Lipid oxidation values significantly increased during maturation process but low levels of malondialdehyde were reported; however, protein oxidation did not significantly change during ripening.     

The effect of mendi (Chaerophyllum sp.) on ripening of vacuum-packed herby cheese

The composition, biochemical and sensory parameters of control cheese (without herbs) and four herby cheeses at 0.5, 1, 2 and 3% herb levels (mendi, Chaerophyllum sp.) ripened at 4 ± 1°C for 90 days were compared. As herb levels increased from 0.5 to 3%, dry matter and pH value decreased significantly. However, dry matter of all cheeses showed similar changes during ripening. The salt content of samples changed from 3.44 to 5.47% during ripening. There was a tendency toward slightly higher titratable acidity in cheeses with more added herbs. Ripening index, trichloroacetic acid-soluble nitrogen/total nitrogen, phosphotungstic acid-soluble nitrogen/total nitrogen, and lipolysis values of the cheese samples were affected by adding herbs and by ripening time. The most acceptable sensory score was obtained with 1% added herbs.     

Lipolysis in cheddar cheese made from raw, thermized, and pasteurized milks

The evolution of free fatty acids (FFA) was monitored over 168 d of ripening in Cheddar cheeses manufactured from good quality raw milk (RM), thermized milk (TM; 65°C × 15 s), and pasteurized milk (PM; 72°C × 15 s). Heat treatment of the milk reduced the level and diversity of raw milk microflora and extensively or wholly inactivated lipoprotein lipase (LPL) activity. Indigenous milk enzymes or proteases from RM microflora influenced secondary proteolysis in TM and RM cheeses. Differences in FFA in the RM, TM, and PM influenced the levels of FFA in the subsequent cheeses at 1 d, despite significant losses of FFA to the whey during manufacture. Starter esterases appear to be the main contributors of lipolysis in all cheeses, with LPL contributing during production and ripening in RM and, to a lesser extent, in TM cheeses. Indigenous milk microflora and nonstarter lactic acid bacteria appear to have a minor contribution to lipolysis particularly in PM cheeses. Lipolytic activity of starter esterases, LPL, and indigenous raw milk microflora appeared to be limited by substrate accessibility or environmental conditions over ripening.     

The evolution of free fatty acids during the ripening of Idiazábal cheese: influence of rennet type

 The object of the present study was to determine the influence of the rennet type on the free fatty acid (FFA) content of Idiazábal cheese. Varying this parameter during cheese-making resulted in significant differences in FFA content in the cheeses during ripening. The main FFAs in both cheese batches were caproic acid (C6 : 0) and capric acid (C10 : 0). The differences between the extreme values for the lipolysis rate were around 45%, which emphasizes the importance of the cheese-making procedure employed on lipolysis in this type of cheese. The use of locally made rennet in the preparation of Idiazábal cheese increased the level of lipolysis in the cheese. Received: 4 December 1998 / Revised version: 18 March 1999     

Evolution of lipolysis during the ripening of traditional Feta cheese

Lipolysis was studied during ripening of traditional Feta cheese produced in two small dairies, A and B. The cheeses were made from a thermized mixture of ewes’/goats’ milk by using yoghurt as starter and artisanal rennet from lambs’ and kids’ abomasa (cheese A) or mixed artisanal rennet with calf rennet (cheese B).The acid degree value and the free fatty acids (FFA) contents in both cheeses increased sharply up to 18 d (pre-ripening period at 15 °C) and continued to increase throughout ripening. In both mature cheeses, acetic acid was found at high levels (13–18% of the total FFAs). However, except for this, all FFA contents differed significantly (P < 0.05) between the two cheeses throughout ripening. The levels of individual and total C2:0–C8:0, C10:0–C14:0 and C16:0–C18:2 fatty acids were significantly higher (P < 0.05) in cheese A than in cheese B. Presumably the difference, especially in the C2:0–C8:0 content, was due mainly to the type of the rennet used. Butyric acid was the dominant FFA in cheese A (20% of the total FFAs at 120 d), while the most abundant FFAs in cheese B were capric (18%) and lauric acid (18%). In general, the lipolysis degree of the two cheeses was higher than those reported for the industrially-made Feta cheese.In organoleptic evaluation, cheese A had a piquant taste that was attributed to its high content of butyric acid and showed a significantly (P < 0.05) higher total score than cheese B.     

Effect of pasture versus indoor feeding regimes on the yield,composition, ripening and sensory characteristics of Maasdam cheese

Maasdam cheese was manufactured from standardized milk derived from each of three feeding systems: grass (GRA), grass and clover pasture (CLO), and indoor feeding of total mixed ration (TMR). Pasture‐derived cheeses had significantly lower L* (whiteness) and higher b* values (yellowness) compared to TMR‐derived cheeses. Acetate levels were significantly lower in CLO and butyrate levels significantly higher in TMR compared to the other cheeses. Grass‐fed cheese had significantly higher scores for smooth texture, ivory colour and shiny appearance compared to TMR. The influence of feed type was minimal on cheese yield, composition and on glycolysis, lipolysis and proteolysis during ripening.     

Evidence of a relationship between autolysis of starter bacteria and lipolysis in cheddar cheese during ripening

Cell viability, autolysis and lipolysis were studied in Cheddar cheese made using Lactococcus lactis subsp. cremoris AM2 or Lactococcus lactis subsp. cremoris HP. Cheddar cheese was made in triplicate over a 3 month period and ripened for 238 days at 8 degrees C. Cell viability in cheese was lower for AM2 (a non-bitter strain) than for strain HP (a bitter strain). Autolysis, monitored by the level of the intracellular marker enzyme, lactate dehydrogenase (EC 1.1.1.27) in cheese 'juice' extracted by hydraulic pressure, was much greater in the cheese made using AM2 than that made with HP. Lipolysis was determined by the increase during ripening of individual free fatty acids (FFA) from butyric (C4:0) to linolenic acid (C18:3) measured using a high performance liquid chromatographic technique. Levels of individual FFA from butyric (C4:0) to linolenic (C18:3) acids increased significantly (P<0.05) during ripening in cheeses made with either starter culture. Palmitic (C16:0) and oleic (C18:1) acids were the most abundant FFA throughout ripening in all cheeses. Levels of caprylic (C8:0), myristic (C14:0), palmitic (C16:0) and stearic (C18:0) acids were significantly higher (P<0.05) in cheeses manufactured with Lc. lactis subsp. cremoris AM2 than in cheeses manufactured with Lc. lactis subsp. cremoris HP. Differences in levels of lipolysis between strains was not due to differences in the specific lipolytic or esterolytic activities in cell free extracts of the strains as measured by activity on triolein (lipase) and p-nitrophenylbutyrate (esterase) substrates. Therefore, evidence is provided for a relationship between the extent of starter cell autolysis and the level of lipolysis during Cheddar cheese ripening.     

Lipolysis and proteolysis profiles of fresh artisanal goat cheese made with raw milk with 3 different fat contents

The objective of this study was to describe the proteolysis and lipolysis profiles in goat cheese made in the Canary Islands (Spain) using raw milk with 3 different fat contents (0.5, 1.5, and 5%) and ripened for 1, 7, 14, and 28 d. β-Casein was the most abundant protein in all cheeses and at all ripening times. Quantitative analysis showed a general decrease in caseins as ripening progressed, and degradation rates were higher for α_S1-casein than for β-casein and α_S2-casein. Furthermore, the degradation rate during the experimental time decreased with lower fat contents. The α_S2-casein and α_S1-casein levels that remained in full-fat and reduced-fat cheeses were less than those in low-fat cheese. In contrast, β-casein also showed degradation along with ripening, but differences in degradation among the 3 cheese types were not significant at 28 d. The degradation products increased with the ripening time in all cheeses, but they were higher in full-fat cheese than in reduced-fat and low-fat cheeses. The free fatty acid concentration per 100 g of cheese was higher in full-fat cheese than in reduced- and low-fat cheese; however, when the results were expressed as milligrams of free fatty acids per gram of fat in cheese, then lipolysis occurred more rapidly in low-fat cheese than in reduced- and full-fat cheeses. These results may explain the atypical texture and off-flavors found in low-fat goat cheeses, likely the main causes of non-acceptance.     

The effects of some starter cultures on the properties of Turkish White cheese

White cheese samples were manufactured from bovine milk using three different commercial direct vat starter cultures (DVS-1, -2 and -3) and a lyophilized culture, and ripened at 4 ± 1°C for 90 days. The composition, titratable acidity and ripening indices of the cheese samples were determined on the 2nd, 30th, 60th and 90th days of ripening. The ratios of total solids, protein and fat were higher for cheeses manufactured using DVS-2 and lyophilized cultures but the titratable acidity in cheese produced using DVS-3 and lyophilized cultures was higher (P < 0.01). The mean value of the ripening indices of the cheese produced using the lyophilized culture was lower than the cheeses produced with added DVS cultures (P < 0.05). The total solids, ash, salt ratios, titratable acidity and ripening indices values increased for all types of white cheeses during ripening (P < 0.05).     

Starter strain related effects on the biochemical and sensory properties of Cheddar cheese

A detailed investigation was undertaken to determine the effects of four single starter strains, Lactococcus lactis subsp. lactis 303, Lc. lactis subsp. cremoris HP, Lc. lactis subsp. cremoris AM2, and Lactobacillus helveticus DPC4571 on the proteolytic, lipolytic and sensory characteristics of Cheddar cheese. Cheeses produced using the highly autolytic starters 4571 and AM2 positively impacted on flavour development, whereas cheeses produced from the poorly autolytic starters 303 and HP developed off-flavours. Starter selection impacted significantly on the proteolytic and sensory characteristics of the resulting Cheddar cheeses. It appeared that the autolytic and/or lipolytic properties of starter strains also influenced lipolysis, however lipolysis appeared to be limited due to a possible lack of availability or access to suitable milk fat substrates over ripening. The impact of lipolysis on the sensory characteristics of Cheddar cheese was unclear, possibly due to minimal differences in the extent of lipolysis between the cheeses at the end of ripening. As anticipated seasonal milk supply influenced both proteolysis and lipolysis in Cheddar cheese. The contribution of non-starter lactic acid bacteria towards proteolysis and lipolysis over the first 8 months of Cheddar cheese ripening was negligible.     

The manufacture of miniature Cheddar-type cheeses from milks with different fat globule size distributions

A novel 2-stage gravity separation scheme was developed for fractionation of raw, whole bovine milk into fractions enriched in small (SFG) or large (LFG) fat globules. The volume mean diameter of fat globules in SFG, LFG or control (CTRL) milk was 3.45, 4.68 and 3.58 microm, respectively. The maximum in storage modulus (index of firmness) decreased with increasing fat globule size for rennet-induced gels formed from SFG, LFG or CTRL milks. Miniature (20 g) Cheddar cheeses were manufactured using each of the 3 milks. There were no significant (P > 0.05) differences in the pH, moisture and fat in dry matter levels between cheeses made using any of the 3 milks, however, the fat content of the cheese made using SFG milk was approximately 1% lower than that of cheese made using LFG or CTRL milk in each of the 2 trials. Image analysis of confocal scanning laser micrographs of the cheeses illustrated that the star volume of fat globules in the cheeses decreased significantly (P < or = 0.05) as the size of fat globules in the milks used for cheesemaking was reduced. This indicates that it is possible to manipulate the size distribution of fat globules in Cheddar cheese by adjusting the fat globule size distribution of the milk used for cheese-making. The concentration of free fatty acids (FFA) increased in all cheeses during ripening. At 120 d of ripening, the concentration of FFA varied significantly (P < or = 0.05 and P < or = 0.001 for trials 1 and 2, respectively) with fat globule size, with cheeses made in trial 2 from LFG, SFG or CTRL milks having total FFA levels of 3391, 2820 and 2612 mg/kg cheese, respectively.     

Effect of calcium on fatty acid bioaccessibility during in vitro digestion of Cheddar-type cheeses prepared with different milk fat fractions

Calcium plays an important role in intestinal lipid digestion by increasing the lipolysis rate, but also limits fatty acid bioaccessibility by producing insoluble Ca soaps with long-chain fatty acids at intestinal pH conditions. The aim of this study was to better understand the effect of Ca on the bioaccessibility of milk fat from Cheddar-type cheeses. Three anhydrous milk fats (AMF) with different fatty acid profiles (olein, stearin, or control AMF) were used to prepare Cheddar-type cheeses, which were then enriched or not with Ca using CaCl₂ during the salting step. The cheeses were digested in vitro, and their disintegration and lipolysis rates were monitored during the process. At the end of digestion, lipids were extracted under neutral and acidic pH conditions to compare free fatty acids under intestinal conditions in relation to total fatty acids released during the digestion process. The cheeses prepared with the stearin (the AMF with the highest ratio of long-chain fatty acids) were more resistant to disintegration than the other cheeses, owing to the high melting temperature of that AMF. The Ca-enriched cheeses had faster lipolysis rates than the regular Ca cheeses. Chromatographic analysis of the digestion products showed that Ca interacted with long-chain fatty acids, producing Ca soaps, whereas no interaction with shorter fatty acids was detected. Although higher Ca levels resulted in faster lipolysis rates, driven by the depletion of reaction products as Ca soaps, such insoluble compounds are expected to reduce the bioavailability of fatty acids by hindering their absorption. These effects on lipid digestion and absorption are of interest for the design of food matrices for the controlled release of fat-soluble nutrients or bioactive molecules.