The behavior of cathepsin D during milk processing and its contribution to bitterness in a model fresh cheese.

The bovine endopeptidase cathepsin D was investigated regarding its temperature-dependent inactivation and ability to form bitter peptides within a spiked model fresh cheese. Cathepsin D was found to be more susceptible than other milk endogenous peptidases to temperature treatments in skim milk. Inactivation kinetics revealed decimal reduction times of 5.6 min to 10 s in a temperature range from 60 to 80°C. High temperature and ultra-high temperature (UHT) treatments from 90 to 140°C completely inactivated cathepsin D within 5 s. A residual cathepsin D activity of around 20% was detected under pasteurization conditions (72°C for 20 s). Therefore, investigations were done to estimate the effect of residual cathepsin D activity on taste in a model fresh cheese. The UHT-treated skim milk was spiked with cathepsin D and acidified with glucono-δ-lactone to produce a model fresh cheese. A trained bitter-sensitive panel was not able to distinguish cathepsin D-spiked model fresh cheeses from the control model fresh cheeses in a triangle test. Model fresh cheese samples were also analyzed for known bitter peptides derived from casein fractions using a HPLC-tandem mass spectrometry (MS) approach. In accordance with the sensory evaluation, the MS analyses revealed that the bitter peptides investigated within the cathepsin D-spiked model fresh cheese were not found or were below the limit of detection. Even though cathepsin D may be present during the fermentation of pasteurized milk, it does not seem to be responsible for bitter peptide formation from milk proteins on its own.

Technologies for sustainable heat generation in food processing.

The utilization of heat is one of the foundations of modern food processing. At present, boilers that operate on fossil fuels are still dominating the generation of hot water, steam, and hot air in the food industry. In light of sustainability goals and carbon taxes as well as international efforts to reduce the dependence on natural gas, new technologies are needed to lower the greenhouse gas emissions related to thermal processing of foods. This review discusses important technologies that could serve as a replacement for conventional fossil fuel boilers in the future. These technologies are based on electricity, geothermal energy (direct/indirect use), and electricity to hydrogen conversion and include fuel cells, microturbines, engines, electrical boilers, heat pumps, radiation, and use of geothermal energy. The majority of these technologies are already available for implementation at larger scales and emissions are generally lower compared to burning fossil fuels. At present, major obstacles, such as low fossil fuel prices, still exist that prevent the widespread adoption of more sustainable heating technologies. However, the direct transformation of electrical energy and utilization of geothermal energy for heating purposes seem promising and should be more frequently installed in the future, whereas the use of H2 obtained through electrolysis as a transportable source of energy may also serve as a source of thermal energy where it is useful to generate electricity and heat on the production site or where the availability of electricity is limited.

Studying dynamic aroma release by headspace-solid phase microextraction-gas chromatography-ion mobility spectrometry (HS-SPME-GC-IMS): method optimization, validation, and application.

To understand aroma perception from complex food matrices' determination of dynamic aroma release during simulated oral processing is necessary. In this study optimization, validation and application of a novel method coupling headspace-solid phase microextraction (HS-SPME) with gas chromatography-ion mobility spectrometry (GC-IMS) is presented. Thirteen character impact compounds imparting different chemical properties are studied to understand capabilities and limitations of the method. It was shown for the first time that the temperature of the IMS sample inlet can be increased up to 200 °C without instrumental constraints. Linear calibration was possible for eleven of the thirteen compounds with one decade dynamic range. The limit of detection and quantitation were 2.1-63.0 ppb and 7.2-210.1 ppb, respectively. Diacetyl could be detected in negative polarity mode of IMS, however with lower precision compared to the compounds detected in positive mode. Limitations of the method were short HS-SPME extraction time, which in the case of caproic acid was not sufficient for reliable quantification. Additionally, δ-decalactone could not be detected due to maximum GC temperature of 200 °C. Application of the method to determine dynamic aroma release from a dairy matrix was successfully shown for nine compounds. Analysis of complex food matrix was performed with similar precision compared to analysis in aqueous solution, thus proving high robustness of the method towards matrix effects.

Behavior of concentrated emulsions prepared by acid-hydrolyzed insoluble microalgae proteins from Chlorella protothecoides.

BACKGROUND: Microalgae are a promising alternative source to meet the increasing global demand for protein. The insoluble microalgae protein fraction that makes up over half of the protein composition of the biomass has shown potential to serve as a functional emulsifier after acidic hydrolysis. However, creaming was observed due to the flocculation of emulsion droplets, suggesting a preferable use in concentrated emulsions. RESULTS: In this study, we examined the emulsifying behavior of the untreated insoluble microalgae protein fraction and two of its hydrolysates obtained in 0.5 mol L-1 HCl for 4 h at 65 °C (Hydrolysates 65) or 85 °C (Hydrolysates 85), at a concentration of 3% (w/w), and elevated levels of oil (50-70%). The results showed an increase in droplet size and apparent viscosity with increasing oil content in the emulsions. The emulsions made with Hydrolysates 85 had the smallest droplet size and the highest apparent viscosity. The gravitational separation was hindered when oil content was increased. The Hydrolysates 85 stabilized emulsions had a gel-like structure and were stable against coalescence or creaming during a 7 day storage test. CONCLUSION: The results suggest that the thermal acid-treated fraction Hydrolysates 85 may, in particular, be a good emulsifier to formulate concentrated emulsion-based foods with oil content over 50%, such as mayonnaise, salad dressings, or dips. © 2020 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Fat-free fermented concentrated milk products as milk protein-based microgel dispersions: Particle characteristics as key drivers of textural properties.

The popularity of fat-free fermented concentrated milk products, such as fresh cheeses and high-protein yogurt, has increased over the recent years, attributed to greater availability and improvements in taste and texture. These improvements have been achieved through modifications and new developments in processing technologies, for example, higher heat treatment intensities and incorporating different membrane filtration technologies. Though numerous processing parameters are discussed in the literature, as well as reasons behind the developments, detailed examinations of how process modifications affect the final textural attributes of these products are lacking. To draw links between processing parameters and texture, we review the literature on fat-free fermented concentrated milk products from the perspective of fermented milk protein-based microgel particles as the basic structural unit. At each main processing step, relationships between process parameters, micro- and macrostructural and sensory (textural) properties are discussed.An overview of particle characteristics that drive structural changes at each processing step is developed in relation to textural characteristics. Using this approach of assessing relationships between structural characteristics of concentrated dispersions of fat-free fermented milk protein-based microgel particles and processing parameters provides a basic context for the selection of optimal parameters to achieve a desired texture.

Cultivation and downstream processing of microalgae and cyanobacteria to generate protein-based technofunctional food ingredients.

Microalgae are unicellular microorganisms that can be rich in proteins and are therefore a valuable ingredient in different foods. So far microalgae are mainly utilized in foods in low concentrations as a whole-cell ingredient even though it is known that proteins extracted from microalgae can possibly posess various technofunctional properties, such as high protein solubility, emulsification, foaming, and gelation properties. The widespread usage of protein-rich ingredients obtained from microalgae is for the most part prevented by the high price of the biomass, the lack of efficient downstream processes, and the adverse taste. The aim of this review is to give insights into the fundamental properties of the growth and processing of microalgae, highlight the advantages of microalgae ingredients and show potential applications based on the technofunctional, nutritional and sensory properties that were reported. Moreover, the existing challenges and knowledge gaps that hinder the application of microalgal proteins in foods are discussed.

Determination of mono- and diacylglycerols from E 471 food emulsifiers in aerosol whipping cream by high-performance thin-layer chromatography-fluorescence detection.

Mono- and diacylglycerol (MAG and DAG) emulsifiers (E 471) are widely applied to regulate techno-functional properties in different food categories, for example, in dairy products. A method for the determination of MAG and DAG in aerosol whipping cream by high-performance thin-layer chromatography with fluorescence detection (HPTLC-FLD) after derivatization with primuline was developed. For sample preparation, aerosol whipping cream was mixed with ethanol, followed by the addition of water and liquid-liquid extraction with tert-butyl methyl ether. The sample extracts were analyzed by HPTLC-FLD on silica gel LiChrospher plates with n-pentane/n-hexane/diethyl ether (22.5:22.5:55, v/v/v) as mobile phase, when interfering matrix like cholesterol and triacylglycerols were successfully separated from the E 471 food additives. For quantitation, an emulsifier with known composition was used as calibration standard and the fluorescent MAG and DAG were scanned at 366/> 400 nm. Limits of detection and quantitation of 4 and 11 mg/100 g aerosol whipping cream were obtained for both monostearin and 1,2-distearin, respectively, and allowed the reliable quantitation of MAG and DAG from E 471 far below commonly applied emulsifier amounts. Recoveries from model aerosol whipping cream with 400 mg E 471/100 g were determined in a calibration range of 200-600 mg E 471/100 g sample and ranged between 86 and 105% with relative standard deviations below 7%. In aerosol whipping creams from the German market, E 471 amounts ranged between 384 and 610 mg/100 g.

Impact of the oil droplet size on the oxidative stability of microencapsulated oil.

Microencapsulation aims to protect polyunsaturated fatty acids against oxidation by embedding oil droplets in a solid matrix. The effect of the oil droplet size on the oxidation was evaluated under consideration of the non-encapsulated oil and the powder particle size. An O/W emulsion (1.6%(w/w) soy protein; 30.4%(w/w) maltodextrin DE21; 8%(w/w) fish oil) was homogenised at different pressure levels (4, 8, 17.5 and 30 MPa). Emulsions were spray dried and the size of the obtained powders was standardised. Powders were stored for 147 days at 25 °C and the hydroperoxide concentration and Anisidine Value in the total- and encapsulated oil measured. The volume mean diameter of oil droplets varied between 0.48 ± 0.01 and 1.54 ± 0.07 µm. Powders containing small oil droplets resulted in fewer oxidation products, which was related to a larger specific surface area and therefore a pronounced chemical stabilisation by soy protein isolate rather than oxygen diffusion phenomena or different encapsulation efficiencies.

Ionic strength and pH stability of oil-in-water emulsions prepared with acid-hydrolyzed insoluble proteins from Chlorella protothecoides.

BACKGROUND: Chlorella protothecoides is one of the most widely commercialized and studied microalgae species. Recent research reported improved emulsifying properties of the insoluble protein fraction from C. protothecoides after thermal-acid treatment. RESULTS: In this research, we studied the influence of ionic strength (sodium chloride 50-500 mmol L-1 or calcium chloride 5-50 m mol L-1 ) and pH (2-9) on the stability of oil-in-water emulsions prepared by 3% (w/w) of the untreated insoluble microalgae protein fraction or hydrolysates obtained after treatment with hydrochloric acid at 65 °C (Hydrolysates 65) or 85 °C (Hydrolysates 85) for 4 h. The emulsions were prepared by mixing 10% (w/w) oil and homogenized at 68.9 MPa. The ionic strength and pH were, subsequently, adjusted. The mean particle diameter of emulsions remained constant despite extensive variations in ionic strength or pH. Emulsion droplets stabilized by Hydrolysates 85 were stable against coalescence at all ionic strengths or pH values tested. CONCLUSION: The results indicate a high potential to use acid-hydrolyzed insoluble microalgae protein fractions for the formulation of various emulsion-based food systems. © 2020 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Sensory properties of aqueous dispersions of protein-rich extracts from Chlorella protothecoides at neutral and acidic pH.

BACKGROUND: Water-soluble proteins extracted from the heterotrophically cultivated microalga Chlorella protothecoides have been shown to have a good solubility over a broad pH range, which makes them a promising candidate for beverage formulations. This study investigated the sensory properties of dispersions of a protein-rich extract from C. protothecoides at neutral and pH 3. RESULTS: Sensory acceptance tests of the pure extract revealed an overall low acceptance at pH 7 without sucrose addition. Sensory acceptance was significantly (P ≤ 0.05) increased by lowering the pH to 3 with citric acid, and the addition of 50 g kg-1 sucrose. Here, overall positive sensory acceptance ratings were achieved up to a protein extract concentration of 40 g kg-1 . Basic taste evaluations showed only low bitterness scores and no significant (P > 0.05) increase in bitterness with decreasing pH. CONCLUSION: It is suggested that protein-rich extracts from C. protothecoides have promising sensory properties in beverage formulations. © 2019 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

An erosion-type hydrolysis behavior of insoluble protein fraction from Chlorella protothecoides.

BACKGROUND: Acid-induced hydrolysis of proteins has been used to improve the solubility and functional properties of various proteins, and could be a promising tool to facilitate the use of currently underutilized insoluble microalgae protein-rich fractions in food applications. However, the results of a prior study showed an unusual resistance of an insoluble microalgae protein-rich fraction to acid hydrolysis at room temperature. RESULTS: In the present study, the insoluble protein-rich fraction extracted from microalgae Chlorella prothothecoides was treated with 0.5 mol L-1 hydrochloric acid at 25, 45, 65 or 85 °C for 0-4 h. The results showed that hydrolysis of the fraction at 85 °C for 4 h led to decreases in the amount of insoluble protein-rich aggregates and the formation of fragments with a lower molecular weight, as well as an increase in protein solubility by approximately 40%. Nevertheless, some aggregated insoluble protein-rich particles remained, even after hydrolysis at 85 °C for 4 h. CONCLUSION: The higher temperature improved the efficiency of the acid hydrolysis of the insoluble protein fraction from microalgae Chlorella prothothecoides, which is highly acid-resistant. Overall, an erosion-based mechanism was suggested for the acid hydrolysis of insoluble microalgae protein fraction. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Resistance of thermophilic spore formers isolated from milk and whey products towards cleaning-in-place conditions: Influence of pH, temperature and milk residues.

The occurrence of thermophilic spore formers in dairy powders is a major concern for producers worldwide. This study aims to investigate the resistance of thermophilic endospores towards cleaning solutions typically used for cleaning-in-place in dairy manufacturing plants. From eleven tested strains, all were able to survive an alkaline treatment (NaOH) at 65 °C for 10 min (0.5%), whereas at concentrations of 2% eight strains withstood the treatment. Acid solutions were more sporicidal. At 0.5% of HNO3, only three strains survived the treatment. Milk impurities reduced the inactivation effect of the NaOH solutions towards thermophilic spore formers. For two selected strains, a detailed kinetic inactivation in NaOH and HNO3 solutions at different temperatures was performed and non-log-linear inactivation curves were observed. This study highlights the risk of reusing cleaning solutions in dairies.

Power ultrasound as a tool to improve the processability of protein-enriched fermented milk gels for Greek yogurt manufacture.

One approach to avoid production of acid whey during the manufacture of high-protein yogurt and related products is to concentrate the milk before fermentation. However, the resultant gels are firm so that stirring in the tank and further processing are difficult on an industrial scale. We hypothesize that power ultrasound (US) during fermentation softens the gel because sound waves cause cavitation and strong shear forces in the fluid. Skim milk was standardized to different protein contents up to 12%, heated (85°C, 30 min), and acidified with thermophilic or mesophilic starter cultures. An excessive increase in gel firmness as a function of protein content was detected. In the next series of experiments, US was applied during fermentation. Milks (10% protein) were acidified at 43.5°C and sonicated from pH 5.8 to 5.1 with a sonotrode (20 kHz, 20 W). Immediately after fermentation, gels were agitated using a rheometer with a vane geometry. The maximum torque required to break the gel was reduced by 75% following US, and gel firmness was reduced by 80%. Gels were then processed into stirred yogurt and analyzed. Sonicated samples were smoother with fewer large aggregates. Confocal laser scanning microscopy images suggested a less cohesive structure and more compact microgel particles, resulting in reduced viscosity. We concluded that US is a promising tool to weaken the gel and facilitate further processing. This enables new approaches for the manufacture of Greek yogurt, particularly in regard to avoiding production of acid whey and developing products with novel textures.

Vibration-induced particle formation during yogurt fermentation-Effect of frequency and amplitude.

Machinery such as pumps used for the commercial production of fermented milk products cause vibrations that can spread to the fermentation tanks. During fermentation, such vibrations can disturb the gelation of milk proteins by causing texture defects including lumpiness and syneresis. To study the effect of vibrations on yogurt structure systematically, an experimental setup was developed consisting of a vibration exciter to generate defined vibrational states and accelerometers for monitoring. During the fermentation of skim milk, vibrations (frequency sweep: 25 to 1,005 Hz) were introduced at different pH (5.7 to 5.1, step width 0.1 units) for 200 s. Physical properties of set gels (syneresis, firmness) and resultant stirred yogurts (visible particles, rheology, laser diffraction) were analyzed. Vibrational treatments at pH 5.5 to 5.2 increased syneresis, gel firmness, and the number of large particles (d > 0.9 mm); hence, this period was considered critical. The particle number increased from 34 ± 5 to 242 ± 16 particles per 100 g of yogurt due to vibrations at pH 5.4. In further experiments, yogurts were excited with fixed frequencies (30, 300, and 1,000 Hz). All treatments increased syneresis, firmness, and particle formation. As the strongest effect was observed by applying 30 Hz, the amplitude was set to vibration accelerations of a = 5, 10, 15, 20, and 25 m/s2 in the final experiments. The number of large particles was increased due to each treatment and a positive correlation with the amplitude was found. We concluded that vibrations during gelation increase the collision probability of aggregating milk proteins, resulting in a compressed set gel with syneresis. Resultant stirred yogurts exhibit large particles with a compact structure leading to a reduced water-holding capacity and product viscosity.

Oil Perception-Detection Thresholds for Varying Fatty Stimuli and Inter-individual Differences.

Multiple lines of research have demonstrated that humans can perceive fat in the form of free fatty acids (FFAs). However, the dietary concentration of FFAs is generally very low and fat is mainly consumed as triacylglycerol (TAG). The aim of this study was to examine the perception of different fatty stimuli and possible associations between them. Therefore, detection thresholds for 4 fatty stimuli (oleic acid [FFA], paraffin oil [mixture of hydrocarbon molecules], canola oil [TAG-rich], and canola oil spiked with oleic acid [rich in TAGs and FFAs]) were determined in 30 healthy participants. Additionally, inter-individual differences in fat perception were examined. It was observed that oleic acid was perceivable at significantly lower concentrations than all other stimuli (P < 0.001). Similarly, canola oil with oleic acid was detectable at lower concentrations than canola oil alone (P < 0.001). Moreover, canola oil detection thresholds were significantly lower than paraffin oil detection thresholds (P = 0.017). Participants who were sensitive for low concentrations for oleic acid showed lower detection thresholds for canola oil with and without oleic acid, compared with participants that were less sensitive for oleic acid. The results of this study demonstrate that the higher the concentrations of FFAs in the stimuli, the lower the individual fat detection threshold. Moreover, participants being sensitive for lower concentrations of FFAs are also more likely to detect low concentrations of TAG-rich fats as it is found in the human diet.

Pseudomonas lactis sp. nov. and Pseudomonas paralactis sp. nov., isolated from bovine raw milk.

Five strains, designated WS 4672T, WS 4998, WS 4992T, WS 4997 and WS 5000, isolated from bovine raw milk formed two individual groups in a phylogenetic analysis. The most similar species on the basis of 16S rRNA gene sequences were Pseudomonas azotoformans IAM 1603T, Pseudomonas gessardii CIP 105469T and Pseudomonas libanensis CIP 105460T showing 99.7-99.6 % similarity. Using rpoD gene sequences Pseudomonas veronii LMG 17761T (93.3 %) was most closely related to strain WS 4672T and Pseudomonas libanensis CIP 105460T to strain WS 4992T (93.3 %). The five strains could be differentiated from their closest relatives and from each other by phenotypic and chemotaxonomic characterization and ANIb values calculated from draft genome assemblies. ANIb values of strains WS 4992T and WS4671T to the closest relatives are lower than 90 %. The major cellular polar lipids of both strains are phosphatidylethanolamine, phosphatidylglycerol, a phospholipid and diphosphatidylglycerol, and their major quinone is Q-9. The DNA G+C content of strains WS 4992T and WS 4672T were 60.0  and 59.7  mol%, respectively. Based on these genotypic and phenotypic traits two novel species of the genus Pseudomonas are proposed: Pseudomonas lactis sp. nov. [with type strain WS 4992T (=DSM 29167T=LMG 28435T) and the additional strains WS 4997 and WS 5000], and Pseudomonasparalactis sp. nov. [with type strain WS 4672T (=DSM 29164T=LMG 28439T) and additional strain WS 4998].

Chemical and optical characterization of white efflorescences on dry fermented sausages under modified atmosphere packaging.

BACKGROUND: Dry fermented sausages that are packed under modified atmosphere are often affected by the formation of white crystals on the surface. These so called efflorescences are rejected by consumers and lead to high financial losses for the meat processing industry. In this study, the distribution of efflorescence-causing components was investigated over the sausage profile during 8 weeks of storage under modified atmosphere at 4 °C. In addition, two visual methods (image and sensory analyses) were compared regarding the ability to quantify the efflorescence content. RESULTS: The initial formation of efflorescences was observed after 2 weeks (7%). After 4 weeks of storage, 23.4% of the sausage surface was covered with efflorescences, and the amount of efflorescences did not change significantly by the end of storage. Furthermore, chemical analyses revealed that magnesium (increased by 98.1%), lactate (increased by 54.2%) and creatine (increased by 51.8%) are enriched on the sausage surface during storage. CONCLUSION: Sensory and image analyses lead to comparable results (r = 0.992) and therefore both are suitable to quantify the amount of efflorescences. The moisture gradient in the interior of the sausages which is built upon drying is supposed to be the driving force for the movement of efflorescence-causing compounds. © 2017 Society of Chemical Industry.

Novel cellobiose 2-epimerases for the production of epilactose from milk ultrafiltrate containing lactose.

A selected number of enzymes have recently been assigned to the emerging class of cellobiose 2-epimerases (CE). All CE convert lactose to the rare sugar epilactose, which is regarded as a new prebiotic. Within this study, the gene products of 2 potential CE genes originating from the mesophilic bacteria Cellulosilyticum lentocellum and Dysgonomonas gadei were recombinantly produced in Escherichia coli and purified by chromatography. The enzymes have been identified as novel CE by sequence analysis and biochemical characterizations. The biochemical characterizations included the determination of the molecular weight, the substrate spectrum, and the kinetic parameters, as well as the pH and temperature profiles in buffer and food matrices. Both identified CE epimerize cellobiose and lactose into the C2 epimerization products glucosylmannose and epilactose, respectively. The epimerization activity for lactose was maximal at pH 8.0 or 7.5 and 40°C in defined buffer systems for the CE from C. lentocellum and the CE from D. gadei, respectively. In addition, biotransformations of the foodstuff milk ultrafiltrate containing lactose were demonstrated. The CE from D. gadei was produced in a stirred-tank reactor (12 L) and purified using an automatic system. Enzyme production and purification in this scale indicates that a future upscaling of CE production is possible. The bioconversions of lactose in milk ultrafiltrate were carried out either in a batch process or in a continuously operated enzyme membrane reactor (EMR) process. Both processes ran at an industrially relevant low temperature of 8°C to reduce undesirable microbial growth. The enzyme was reasonably active at the low process temperature because the CE originated from a mesophilic organism. An epilactose yield of 29.9% was achieved in the batch process within 28 h of operation time. In the continuous EMR process, the epilactose yield in the product stream was lower, at 18.5%. However, the enzyme productivity was approximately 6 times higher because the continuous epilactose formation was carried out for about 6 d without further addition of biocatalyst. Within this time, 24g of epilactose in 2.8 L of permeate were produced. The batch and the EMR process showed that the milk ultrafiltrate, which is a sidestream of the milk protein production, might be upgraded to a dairy product of higher value by the enzymatic in situ production of epilactose.

Formation of Key Aroma Compounds During 30 Weeks of Ripening in Gouda-Type Cheese Produced from Pasteurized and Raw Milk.

Gouda-type cheeses were produced on a pilot-scale from raw milk (RM-G) and pasteurized milk (PM-G). Sixteen key aroma compounds previously characterized by the sensomics approach were quantitated in the unripened cheeses and at five different ripening stages (4, 7, 11, 19, and 30 weeks) by means of stable isotope dilution assays. Different trends were observed in the formation of the key aroma compounds. Short-chain free fatty acids and ethyl butanoate as well as ethyl hexanoate continuously increased during ripening but to a greater extent in RM-G. Branched-chain fatty acids such as 3-methylbutanoic acid were also continuously formed and reached a 60-fold concentration after 30 weeks, in particular in PM-G. 3-Methylbutanal and butane-2,3-dione reached a maximum concentration after 7 weeks and decreased with longer ripening. Lactones were high in the unripened cheeses and increased only slightly during ripening. Recent results have shown that free amino acids were released during ripening. The aroma compounds 3-methylbutanal, 3-methyl-1-butanol, and 3-methylbutanoic acid are suggested to be formed by microbial enzymes degrading the amino acid l-leucine following the Ehrlich pathway. To gain insight into the quantitative formation of each of the three aroma compounds, the conversion of the labeled precursors (13C6)-l-leucine and (2H3)-2-keto-4-methylpentanoic acid into the isotopically labeled aroma compounds was studied. By applying the CAMOLA approach (defined mixture of labeled and unlabeled precursor), l-leucine was confirmed as the only precursor of the three aroma compounds in the cheese with the preferential formation of 3-methylbutanoic acid.

Influence of Milk Pasteurization on the Key Aroma Compounds in a 30 Weeks Ripened Pilot-Scale Gouda Cheese Elucidated by the Sensomics Approach.

Gouda cheese was produced from pasteurized milk and ripened for 30 weeks (PM-G). By application of gas chromatography/olfactometry and an aroma extract dilution analysis on the volatiles isolated by extraction/SAFE distillation, 25 odor-active compounds in the flavor dilution (FD) factor range from 16 to 4096 were identified. Butanoic acid, 2- and 3-methylbutanoic acid, and acetic acid showed the highest FD factors, and 2-phenylethanol, δ-decalactone, and δ-dodecalactone were most odor-active in the neutral-basic fraction. Quantitations by stable isotope dilution assays followed by a calculation of odor activity values (OAVs) revealed acetic acid, 3-methylbutanoic acid, butanoic acid, and butane-2,3-dione with the highest OAVs. Finally, an aroma recombinate prepared based on the quantitative data well agreed with the aroma profile of the PM-G. In Gouda cheese produced from raw (nonpasteurized) milk (RM-G), qualitatively the same set of odor-active compounds was identified. However, higher OAVs of butanoic acid, hexanoic acid, and their corresponding ethyl esters were found. On the other hand, in the PM-G, higher OAVs for 3-methylbutanoic acid, 3-methylbutanol, 3-methylbutanal, and butane-2,3-dione were determined. The different rankings of these key aroma compounds clearly reflect the aroma differences of the two Gouda-type cheeses. A higher activity of lipase in the RM-G and higher amounts of free l-leucine in PM-G on the other side were responsible for the differences in the concentrations of some key aroma compounds.