Correlation of the Flavor Characteristics of Swiss-Type Cheeses with Chemical Parameters

Fifteen Swiss-type cheeses were evaluated by a flavor profile method. The cheeses also were analyzed for free fatty acids in whole cheese and in the oil phase and for proteolysis, pH, carbonyls, and gross composition. The flavor notes and chemical parameters were grouped by factor analysis and correlated. Factor analysis showed that many of the free fatty acids varied together. The free fatty acid groups consisted of normal short-chain fatty acids (C₄ to C₁₀), long-chain fatty acids (C₁₂ to C₁₈), and branched short-chain and aromatic acids. Many of the flavor notes also were correlated. Characteristic Swiss cheese flavor notes were correlated with low pH, lipolysis, and acetic and propionic acids. Other flavors were negatively correlated with the oil to cheese distribution of many of the free fatty acids and positively correlated with pH, salt concentration, proteolysis, moisture, branched and aromatic acids, and carbonyls.     

Profiles of Organic Acid and Volatile Compounds in Acid-Type Cheeses Containing Herbs and Spices (Surk Cheese)

A study was conducted to evaluate the basic chemical composition, organic acids, volatile compound profiles, and overall acceptability of Surk cheese (acid cheese). The organic acids were determined by reverse phase high performance liqued chromatography method, and volatile compounds were analyzed by static headspace/gas chromatography/mass spectrometry technique. A total of 134 volatile compounds, including 42 esters, 40 terpenes, 15 alcohos, 11 free fatty acids, 6 ketones, 5 aldehydes, 4 alkenes, 4 phenyl propanoids, 3 phenolics, and 4 other compounds, were identified in the Surk cheeses. The main compounds were found to be carvacrol, γ-terpinene, p-cymene, hexanoic acid, octanoic acid, decanoic acid, butanoic acid, and eugenol. The mean total organic acid content of the Surk cheese was 1.71 g/100 g. The main organic acid in the Surk cheese was lactic acid (1067 mg/100 g), followed by acetic, propionic, oxalic, formic, citric, pyruvic, orotic, hippuric, and uric acids.     

High Performance Liquid Chromatographic Determination of Organic Acids in Dairy Products

A simple isocratic HPLC technique was developed for the quantitative analysis of organic acids in dairy products. An Ammex HPX- 87 column at 65°C, 0.0090N H₂SO₄ mobile phase and UV detection at 220 and 275 nm were utilized. Orotic, citric, pyruvic, lactic, uric, formic, acetic, propionic, butyric, and hippuric acids were quantitated for whole milk, skim powder, cultured buttermilk, sour cream, cottage cheese, yogurt, sharp Cheddar cheese, and blue cheese. Over 90% recoveries of acids added to whole milk were observed for ail acids except butyrid; the average recovery for butyric was 86%.     

Flavour precursor development in Cheddar cheese due to lactococcal starters and the presence and lysis of Lactobacillus helveticus.

The rapid release of intracellular enzymes due to autolysis of lactic acid bacteria in the cheese matrix has been shown to accelerate cheese ripening. The objective of this work was to investigate the evolution of the flavour precursors, individual free amino acids (FAAs), free fatty acids (FFAs) and volatile compounds that contribute to the sensory profiles of cheeses at 2, 6 and 8 months of ripening in Cheddar cheese manufactured using starter systems which varied with respect to their autolytic properties. Starter system A contained a blend of two commercial Lactococcus lactis strains (223 and 227) which had a low level of autolysis. System B was identical to A but also included a highly autolytic strain of Lactobacillus helveticus (DPC4571). System C contained only strain DPC4571. Levels of all individual FAAs were elevated in cheeses B and C relative to A after 2 months of ripening. By 8 months of ripening the main FAA were glutamate, leucine, lysine, serine, proline and valine. Levels of C_6:0, C_8:0, C_12:0 and C_18:0 fatty acids did not vary greatly over ripening, while levels of C_4:0, C_10:0, C_14:0, C_16:0 and C_18:1 were elevated in cheeses B and C. Principal component analysis of the headspace volatiles separated cheese A from cheeses B and C. Cheeses B and C had highest levels of dimethyl disulphide, carbon sulphide, heptanal, dimethyl sulphide, ethyl butanoate, 2-butanone, and 2-methyl butanal and were described as having a ‘caramel’ odour and ‘sweet’, ‘acidic’ and ‘musty’ flavour. Cheese A had highest levels of 2-butanol, 2-pentanone, 2-heptanone, 1-hexanol and heptanal and was described as having a ‘sweaty/ sour’ odour and ‘soapy’, ‘bitter’ and ‘mouldy’ flavour. The results highlight the impact of starter lactococci on flavour precursor development and the positive effect of Lb. helveticus and the lysis of this strain on enhancing levels of substrate and flavour precursors early during ripening resulting in early flavour development.     

Acidic components of boar preputial fluid

Several acids present in the acidic fraction of boar preputial fluid have been identified by gas chromatography. They are acetic, propionic, n-butyric, isobutyric, n-valeric, isovaleric, α- and β-methylvaleric, caproic, n-nonanoic and n-undecanoic acids. Unsaturated aliphatic acids include pent-4-enoic, pent-2-enoic, hept-2-enoic and oct-2-enoic acids. Benzoic, phenylacetic and β-phenylpropionic acids were also identified. Several peaks remained unidentified. Quantitative data for the major components (aromatic acids and acetic acid) are presented and also for the C₃ to C₅ saturated aliphatic acids which are known to have unpleasant odours and low thresholds of detection. The contribution of these compounds to the odour of a boar is discussed, but it is concluded that they contribute little to the taint of heated boar fat.     

Effect of added α-ketoglutaric acid,pyruvic acid or pyridoxal phosphate on proteolyis and quality of Cheddar cheese

Cheddar cheese curds were supplemented with 1, 5 or 20 g of α-ketoglutarate or pyruvic acid or 1.2 g pyridoxal-5¹-phosphate/kg cheese curd. The higher levels of keto-acids (5 or 20 g/kg curd) caused undesirable changes in the physico-chemical properties of resultant cheese. All levels of α-ketoglutarate reduced the pH of the cheese and promoted syneresis during pressing, while pyruvic acid increased the pH of the cheese. The numbers of starter and non-starter lactic acid bacteria were not affected by the addition of keto-acids or pyridoxal-5¹-phosphate. α-Ketoglutarate or pyruvic acid, at 1 g/kg, or pyridoxal-5¹-phosphatase, at 1.2 g/kg cheese curd, did not influence primary proteolysis in the cheese. The highest and lowest concentrations of total and individual free amino acids were found in the cheeses treated with pyruvic acid or α-ketoglutarate, respectively. The concentrations of most amino acids were lower in the cheeses treated with pyridoxal-5¹-phosphate than in the control. The results of this study suggest that α-ketoglutarate and pyridoxal-5¹-phosphate enhanced the degradation of most amino acids in Cheddar cheese while pyruvic acid promoted the formation of amino acids. The cheeses treated with α-ketoglutarate were more mature than the control cheese of the same age while pyruvic acid-treated cheese had a better flavour than the control.     

Gas production by Paucilactobacillus wasatchensis WDCO4 is increased in Cheddar cheese containing sodium gluconate

Paucilactobacillus wasatchensis can use gluconate (GLCN) as well as galactose as an energy source and because sodium GLCN can be added during salting of Cheddar cheese to reduce calcium lactate crystal formation, our primary objective was to determine if the presence of GLCN in cheese is another risk factor for unwanted gas production leading to slits in cheese. A secondary objective was to calculate the amount of CO₂ produced during storage and to relate this to the amount of gas-forming substrate that was utilized. Ribose was added to promote growth of Pa. wasatchensis WDC04 (P.waWDC04) to high numbers during storage. Cheddar cheese was made with lactococcal starter culture with addition of P.waWDC04 on 3 separate occasions. After milling, the curd was divided into six 10-kg portions. To the curd was added (A) salt, or salt plus (B) 0.5% galactose + 0.5% ribose (similar to previous studies), (C) 1% sodium GLCN, (D) 1% sodium GLCN + 0.5% ribose, (E) 2% sodium GLCN, (F) 2% sodium GLCN + 0.5% ribose. A vat of cheese without added P.waWDC04 was made using the same milk and a block of cheese used as an additional control. Cheeses were cut into 900-g pieces, vacuum packaged and stored at 12°C for 16 wk. Each month the bags were examined for gas production and cheese sampled and tested for lactose, galactose and GLCN content, and microbial numbers. In the control cheese, P.waWDC04 remained undetected (i.e., <10⁴ cfu/g), whereas in cheeses A, C, and E it increased to 10⁷ cfu/g, and when ribose was included with salting (cheeses B, D, and F) increased to 10⁸ cfu/g. The amount of gas (measured as headspace height or calculated as mmoles of CO₂) during 16 wk storage was increased by adding P.waWDC04 into the milk, and by adding galactose or GLCN to the curd. Galactose levels in cheese B were depleted by 12 wk while no other cheeses had residual galactose. Except for cheese D, the other cheeses with GLCN added (C, E and F) showed little decline in GLCN levels until wk 12, even though gas was being produced starting at wk 4. Based on calculations of CO₂ in headspace plus CO₂ dissolved in cheese, galactose and GLCN added to cheese curd only accounted for about half of total gas production. It is proposed that CO₂ was also produced by decarboxylation of amino acids. Although P.waWDC04 does not have all the genes for complete conversion and decarboxylation of the amino acids in cheese, this can be achieved in conjunction with starter culture lactococcal. Adding GLCN to curd can now be considered another confirmed risk factor for unwanted gas production during storage of Cheddar cheese that can lead to slits and cracks in cheese. Putative risk factors now include having a community of bacteria in cheese leading to decarboxylation of amino acids and release of CO₂ as well autolysis of the starter culture that would provide a supply of ribose that can promote growth of Pa. wasatchensis.     

Identification of free fatty acids and some other volatile flavour compounds from Swiss cheese using on-line supercritical fluid extraction-gas chromatography

On-line supercritical CO₂ extraction - gas chromatography was applied to the isolation and identification of free fatty acids and other volatile compounds of young and ripe Swiss cheese (Emmental) produced in Finland. Extractions were carried out using a micro-cartridge at 40 °C temperature and 10 MPa pressure and the volatile fractions were analysed by DB-WAX column (polyethylene glycol phase) with flame ionization detection and mass spectrometry. The total time of analysis was less than 2.5 h. Acetic acid and propionic acid predominated over the C12–C18 acids, the longer chain fatty acids increasing in concentration during the ripening of the cheese. Due to the high proportions of fatty acids, further fractionation is required for analysis of the less abundant aroma compounds such as alcohols, carbonyls and lactones.     

Texture, proteolysis and viable lactic acid bacteria in commercial cheddar cheeses treated with high pressure

High pressure processing was investigated for controlling Cheddar cheese ripening. One-month-or 4-month-old Cheddar cheeses were subjected to pressures ranging from 200 to 800 MPa for 5 min at 25 C. The number of viable Lactococcus lactis (starter) and Lactobacillus (nonstarter) cells decreased as pressure increased. Subsequent storage of the control and pressure-treated cheeses at 10 degrees C caused viable cell counts to change in some cases. Free amino acid content was monitored as an indicator of proteolysis. Cheeses treated with pressures > or = 400 MPa evolved free amino acids at significantly lower rates than the control. No acceleration in free amino acid development was observed at lower pressures. Pressure treatment did not accelerate the rate of textural breakdown compared with the non-pressure treated control. On the contrary, pressure treatment at 800 MPa reduced the time-dependent texture changes. Results indicate that high pressure may be useful in arresting Cheddar cheese ripening.     

Evolution of Free Amino Acids during the Ripening of Cheddar Cheese Containing Added Lactobacilli Strains

Cheddar cheese was produced with different lactobacilli strains added to accelerate ripening. The concentration of proteolytic products was determined as free amino acids in the water-soluble fraction at two, four, seven and nine months of aging and at two different maturation temperatures (6°C, 15°C). All amino acids increased during ripening and were higher in the Lactobacillus- added cheeses than in the control cheese, and higher in cheeses ripened at 15°C than at 6°C. Glutamic acid, leucine, phenylalanine, valine and lysine were generally in higher proportion in all cheeses. The cheeses with added L. casei-casei L2A were classified as having a “strong Cheddar cheese” flavor after only seven months of ripening at 6°C.     

Tyrosine and phenylalanine catabolism by Lactobacillus cheese flavor adjuncts

Bacterial metabolism of Tyr and Phe has been associated with the formation of aromatic compounds that impart barny-utensil and floral off-flavors in cheese. In an effort to identify possible mechanisms for the origin of these compounds in Cheddar cheese, we investigated Tyr and Phe catabolism by Lactobacillus casei and Lactobacillus helveticus cheese flavor adjuncts under simulated Cheddar cheese-ripening (pH 5.2, 4% NaCl, 15 degrees C, no sugar) conditions. Enzyme assays of cell-free extracts indicated that L. casei strains catabolize Tyr and Phe by successive, constitutively expressed transamination and dehydrogenation reactions. Similar results were obtained with L. helveticus strains, except that the dehydrogenase enzymes were induced during incubation under cheese-ripening conditions. Micellar electrokinetic capillary chromatography of supernatants from L. casei and L. helveticus strains incubated under simulated cheese-ripening conditions confirmed that Tyr and Phe transamination and dehydrogenation pathways were active in both species and also showed these reactions were reversible. Major products of Tyr catabolism were phydroxy phenyl lactic acid and p-hydroxy phenyl acetic acid, while Phe degradation gave rise to phenyl lactic acid, phenyl acetic acid, and benzoic acid. However, some of these products were likely formed by nonenzymatic processes, since spontaneous chemical degradation of the Tyr intermediate p-hydroxy phenyl pyruvic acid produced p-hydroxy phenyl acetic acid, p-hydroxy phenyl propionic acid, and p-hydroxy benzaldehyde, while chemical degradation of the Phe intermediate phenyl pyruvic acid gave rise to phenyl acetic acid, benzoic acid, phenethanol, phenyl propionic acid, and benzaldehyde.     

Size-Exclusion HPLC Separation of Bitter and Astringent Fractions from Cheddar Cheese Made with Added Lactobacillus Strains to Accelerate Ripening

Lactobacillus strains were added as an adjunct to the regular lactic starter in Cheddar cheese manufacture in order to accelerate ripening. Microbial cheese proteolysis resulted in the release of free amino acids which were extracted with the astringent and bitter fractions and separated by size-exclusion and reversed-phase HPLC chromatography. Lactobacillus strains generally increased the degree of proteolysis. L. plantarum and L. brevis produced off-flavors possibly due to an accumulation of medium-size peptides. The control cheese (without lactobacilli) had the most peptides with a mean molecular- weight of < 1000 daltons and had a flavor described as slightly bitter. Addition of L. casei-casei L2A accelerated ripening and yielded a well-aged Cheddar cheese without any bitterness even after 7 months at 6°C.     

Application of Fourier transform infrared spectroscopy for monitoring short-chain free fatty acids in Swiss cheese

Short-chain free fatty acids (FFA) are important sources of cheese flavor and have been reported to be indicators for assessing quality. The objective of this research was to develop a simple and rapid screening tool for monitoring the short-chain FFA contents in Swiss cheese by using Fourier transform infrared spectroscopy (FTIR). Forty-four Swiss cheese samples were evaluated by using a MIRacle three-reflection diamond attenuated total reflectance (ATR) accessory. Two different sampling techniques were used for FTIR/ATR measurement: direct measurement of Swiss cheese slices (∼0.5 g) and measurement of a water-soluble fraction of cheese. The amounts of FFA (propionic, acetic, and butyric acids) in the water-soluble fraction of samples were analyzed by gas chromatography-flame ion-ization detection as a reference method. Calibration models for both direct measurement and the water-soluble fraction of cheese were developed based on a cross-validated (leave-one-out approach) partial least squares regression by using the regions of 3,000 to 2,800, 1,775 to 1,680, and 1,500 to 900 cm⁻¹ for short-chain FFA in cheese. Promising performance statistics were obtained for the calibration models of both direct measurement and the water-soluble fraction, with improved performance statistics obtained from the water-soluble extract, particularly for propionic acid. Partial least squares models generated from FTIR/ATR spectra by direct measurement of cheeses gave standard errors of cross-validation of 9.7 mg/100 g of cheese for propionic acid, 9.3 mg/100 g of cheese for acetic acid, and 5.5 mg/100 g of cheese for butyric acid, and correlation coefficients >0.9. Standard error of cross-validation values for the water-soluble fraction were 4.4 mg/100 g of cheese for propionic acid, 9.2 mg/100 g of cheese for acetic acid, and 5.2 mg/100 g of cheese for butyric acid with correlation coefficients of 0.98, 0.95, and 0.92, respectively. Infrared spectroscopy and chemometrics accurately and precisely predicted the short-chain FFA content in Swiss cheeses and in the water-soluble fraction of the cheese.     

Assessment of Cheddar Cheese Quality by Chromatographic Analysis of Free Amino Acids and Biogenic Amines

The free amino acids and biogenic amines extracted from normal and late-gassing Cheddar cheeses were derivatized with heptafluorobutyric anhydride and trifluoroacetic anhydride, respectively, before quantification by gas-liquid chromatography. On a microgram scale, twenty amino acids were positively identified in both types of cheese, but only high levels of γ-amino acid butyrate (0.3 to 19.4 mg/g) and small quantities of arginate were found to be associated with “poorly aged” Cheddar cheeses. Histamine (1.54 and 1.22 mg/g) and tyramine (0.32 and 0.43 mg/g) were the bioamines present in highest concentrations in both cheeses.     

Dynamic Headspace Analyses of Volatile Compounds of Cheddar and Swiss Cheese during Ripening

The volatile compounds of Cheddar and Swiss cheeses during ripening for 9 wks at 11°and 21°C, respectively, were analyzed by a dynamic headspace analyzer/gas chromatograph every week. The compounds were identified by a combination of retention times and mass spectra. The volatile compounds of Cheddar increased 5.6 and Swiss cheese 15 times as ripening increased from 0 to 9 wks. The amount of volatile compounds of Swiss cheese was 2.6 times greater than that of Cheddar cheese during ripening. The volatile compounds were ketones, alcohols, aldehydes, esters, acids, sulfur compounds, benzenes, and hydrocarbons. Ketones and alcohols accounted for 92% of volatiles from Cheddar cheese and 88% of those from Swiss cheese.     

Effect of Free or Encapsulated Recombinant Aminopeptidase of Lactobacillus rhamnosus S93 on Acceleration of Cheddar Cheese Ripening

The use of recombinant aminopeptidase (PepN) from Lactobacillus rhamnosus S93 in free or encapsulated form was investigated to shorten the duration of Cheddar cheese ripening. Proteolysis was determined by measuring the soluble nitrogen as phosphotungstic acid (PTA-N) derivatives and free amino acids (FAA) over a 6-month period. The experimental cheeses received higher scores for sensory properties than the control cheese. The amounts of PTA-N and total FAA in the cheese with the encapsulated enzyme after 2 months of ripening were close to those of the control cheese after 6 months, suggesting the acceleration in proteolysis by about 4 months.     

Study of taste-active compounds in the water-soluble extract of mature Cheddar cheese

The chemical composition of the water-soluble extracts of mature Cheddar cheese were identified, with the emphasis on understanding the interplay of compounds contributing to the savoury taste in Cheddar. The ultra-filtered water-soluble extracts of two mature Cheddar cheeses were fractionated by gel permeation chromatography (GPC). By sensory evaluation, two taste-active GPC fractions were identified from each cheese. On the basis of chemical profiling of these fractions, aqueous model tastant mixtures were prepared and sensory omission tests carried out. Glutamic acid, organic acids and mineral salts were the main tastants, whereas the other amino acids had a limited impact on taste. The characteristic umami taste was explained by a synergistic effect of glutamic acid and salts. Matching umami taste intensities were obtained from different concentrations of glutamic acid and salts. Unmasking of a bitter or sweet taste from mixtures of sub-threshold concentrations of amino acids without glutamic acids was also observed.     

Autolysis of selected Lactobacillus helveticus adjunct strains during Cheddar cheese ripening

Cheddar cheeses were manufactured on a pilot scale (500 L vats) with three different Lactobacillus helveticus strains, which showed varying degrees of autolysis, added as adjuncts to the starter. Autolysis of adjunct strains was monitored by reduction in cell numbers, level of intracellular enzymes released into the cheese, and by the consequent changes in the degree of proteolysis and concentration of free amino acids in the cheese. The flavour profiles of the cheeses at 6 months were also determined. Significant variation in viability of the Lb. helveticus strains, which showed a positive correlation with the indicators of autolysis, was observed. However, cheese manufactured with the most autolytic strain did not receive the highest flavour scores. The results indicate that whereas autolysis of adjunct strains is an important factor in Cheddar cheese flavour development, other factors also contribute to the overall flavour improvement observed.     

Effect of free and encapsulated recombinant aminopeptidase on proteolytic indices and sensory characteristics of Cheddar cheese

The recombinant aminopeptidase of Lactobacillus rhamnosus S93 in free or encapsulated form was used in production of Cheddar cheese. The effects of these enzymes on the proteolytic indices as well as sensory characteristics have been investigated during Cheddar cheese ripening. An extrusion method was used to encapsulate the enzyme in alginate beads coated with chitosan. The free or encapsulated aminopeptidase were added at the renneting or salting stage at three different concentrations (50, 500, 2000 units per 18 L of milk). Indices of secondary proteolysis were enhanced by increasing the enzyme concentration. Cheeses with the highest concentration of the encapsulated enzyme had significantly higher concentrations of soluble nitrogen in phosphotungstic acid and total free amino acids and received the highest mean scores for the sensory characteristics.