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PubMed2004

Mould contamination in production of semi-hard cheese.

Air, equipment, plastic film, brine and milk were sampled from four cheese factories in Norway during the period September 1997 to May 1999 in order to identify the critical points for mould contamination in the production process. Altogether, 672 samples were collected. Penicillium brevicompactum was the most frequently isolated species from three of the factories, while Geotrichum candidum was the most frequently isolated species from the fourth. P. commune, P. palitans, P. solitum and P. roqueforti ss. roqueforti, all common contaminants on cheese, were found in samples of air and equipment, and the former was also isolated from plastic film. The results in the present study showed that the mould levels in the cheese factories varied between the different control points. The mould levels at some of the air control points had high mould counts while the mould levels in milk and brine, on equipment and on plastic film, generally were low. The fungi at the control points with high mould levels consisted of common cheese contaminants as well as species not commonly isolated from cheese. The statistical analysis showed that air was the major source of the important cheese contaminants P. commune and P. palitans during the production process. High quality air with low number of cheese contaminants in production rooms, especially the wrapping room, is important in order to reduce mould contamination.

Air Microbiology↗
PubMed2004

Enhancement of 2-methylbutanal formation in cheese by using a fluorescently tagged Lacticin 3147 producing Lactococcus lactis strain.

The amino acid conversion to volatile compounds by lactic acid bacteria is important for aroma formation in cheese. In this work, we analyzed the effect of the lytic bacteriocin Lacticin 3147 on transamination of isoleucine and further formation of the volatile compound 2-methylbutanal in cheese. The Lacticin 3147 producing strain Lactococcus lactis IFPL3593 was fluorescently tagged (IFPL3593-GFP) by conjugative transfer of the plasmid pMV158GFP from Streptococcus pneumoniae, and used as starter in cheese manufacture. Starter adjuncts were the bacteriocin-sensitive strains L. lactis T1 and L. lactis IFPL730, showing branched chain amino acid aminotransferase and alpha-keto acid decarboxylase activity, respectively. Adjunct strains were selected to complete the isoleucine conversion pathway and, hence, increase formation of 2-methylbutanal conferring aroma to the cheese. The non-bacteriocin-producing strain L. lactis IFPL359-GFP was included as starter in the control batch. Fluorescent tagging of the starter strains allowed their tracing in cheese during ripening by fluorescence microscopy and confocal scanning laser microscopy. The bacteriocin produced by L. lactis IFPL3593-GFP enhanced lysis of the adjuncts with a concomitant increase in isoleucine transamination and about a two-fold increase of the derived volatile compound 2-methylbutanal. This led to an enhancement of the cheese aroma detected by a sensory panel. The improvement of cheese flavour and aroma may be of significant importance for the dairy industry.

Aldehydes↗
PubMed2004

Survey of bacterial proteins released in cheese: a proteomic approach.

During the ripening of Emmental cheese, the bacterial ecosystem confers its organoleptic characteristics to the evolving curd both by the action of the living cells, and through the release of numerous proteins, including various types of enzymes into the cheese when the cells lyse. In Emmental cheese these proteins can be released from thermophilic lactic acid bacteria used as starters like Lactobacillus helveticus, Lb delbruecki subsp. lactis and Streptococcus salivarius subsp. thermophilus and ripening bacteria such as Propionibacterium freudenreichii. The aim of this study was to obtain a proteomic view of the different groups of proteins within the cheese using proteomic tools to create a reference map. A methodology was therefore developed to reduce the complexity of cheese matrix prior to 2D-PAGE analysis. The aqueous phase of cheese was prefractionated by size exclusion chromatography, bacterial and milk proteins were separated and subsequently characterised by mass spectrometry, prior to peptide mass fingerprint and sequence homology database search. Five functional groups of proteins were identified involved in: (i) proteolysis, (ii) glycolysis, (iii) stress response, (iv) DNA and RNA repair and (v) oxidoreduction. The results revealed stress responses triggered by thermophilic lactic acid bacteria and Propionibacterium strains at the end of ripening. Information was also obtained regarding the origin and nature of the peptidases released into the cheese, thus providing a greater understanding of casein degradation mechanisms during ripening. Different peptidases arose from St thermophilus and Lb helveticus, suggesting that streptococci are involved in peptide degradation in addition to the proteolytic activity of lactobacilli.

Autolysis↗
PubMed2005

Occurrence and ribotypes of Listeria monocytogenes in Gorgonzola cheeses.

In this study 1656 Gorgonzola cheeses, collected from October 2003 to April 2004 in the same industrial plant located in Lombardia (Italy), were analysed in order to evaluate their level of contamination with Listeria monocytogenes after packaging, as well as at the end of the shelf life. A subset of Gorgonzola isolates was submitted to automated EcoRI ribotyping to evaluate their DUP-IDs (DuPont identification library code) and their level of genetic diversity. The isolate ribotyping profiles were included in an on-line database named PathogenTracker database. The strain DUP-IDs and the similarities between Gorgonzola isolates and PathogenTracker human sporadic strains allowed to evaluate the potential virulence of Gorgonzola-associated strains. The L. monocytogenes detection rates observed in the cheese samples monitored after packaging and at the end of the shelf life were 2.1% and 4.8%, respectively. Seventy percent of the strains genotyped were classified in the same ribotype, labelled as 204 S5, indicating that L. monocytogenes population associated to Gorgonzola cheese shows a low level of genetic diversity. Ninety percent of the strains were classified in DUP-IDs belonging to the II pathogenicity lineage identified for L. monocytogenes. That lineage includes serotype 1/2a, 1/2c and 3c strains, associated to the 35% of the human sporadic isolates described in the literature as causing listeriosis. Moreover, 16.7% of Gorgonzola isolates showed a similarity >or=99% with PathogenTracker human sporadic strains. The results of this study showed that the incidence of L. monocytogenes in Gorgonzola cheeses commercialised by the plant tested was lower than that observed in other Italian blue-veined cheeses by other authors. However, it increased during cheese storage and it become double at the end of the cheese shelf life, ranging from 30 to 60 days after packaging.

Cheese↗
PubMed2005

An outbreak of food-borne listeriosis due to cheese in Japan, during 2001.

Food-borne outbreaks caused by Listeria monocytogenes have been recognized in US and European countries. Only sporadic cases, of neonatal listeriosis, have been reported in Japan. Since L. monocytogenes has been often isolated from foods in Japan, food-borne outbreaks potentially could have occurred. In February 2001, L. monocytogenes serotype 1/2b was isolated from a washed-type cheese during routine Listeria monitoring of 123 domestic cheeses. Further samples from products and the environments at the plant that produced the contaminated cheese were examined for L. monocytogenes. L. monocytogenes serotype 1/2b was detected in 15 cheese samples, at most probable number that ranged from <30 to 4.6 x 10(9)/100 g, and in environmental samples. Studies with people who had consumed cheese from the plant revealed 86 persons who had been infected with L. monocytogenes. Thirty-eight of those people had developed clinical symptoms of gastroenteritis or the common cold type after the consumption of cheese. Isolates from those patients exhibited the same serotype, pathogenicity for mice and HeLa cells, DNA fingerprinting patterns and PCR amplification patterns. From the epidemiological and genetic evidence, it appeared that the outbreak was caused by cheese. This is the first documented incidence of food-borne listeriosis in Japan.

Cheese↗
PubMed2006

Control of Listeria monocytogenes in raw-milk cheeses.

The development of Listeria monocytogenes in cheeses made with raw-milk originating from six different farms and according to the Saint-Nectaire cheesemaking technology was studied. Milk was inoculated with two strains of L. monocytogenes at 5 to 10 CFU/25 ml. Microbial and chemical analyses were carried out at appropriate intervals during ripening. L. monocytogenes did not grow in the cores of cheeses prepared with milk originating from three farms. That inhibition could be partially attributed to the pH values and L-lactate content. There was no growth in cheeses with pH below 5.2 and lactate content around 14 mg/g. In all cheeses, L. monocytogenes stopped growing in the cores of cheeses after eight days and some other factors may be involved in the inhibition. No relation was found between L. monocytogenes count and other microbial counts. Growth occurred on cheese surfaces between eight and eighteen days, when the pH significantly increased. The lowest L. monocytogenes growth was found on the surface of cheeses with the lowest pH and without any core growth. Further studies will be performed to clarify the involvement of the microbial community in L. monocytogenes inhibition, in particular during the ripening period.

Cheese↗
PubMed2006

Optimization of "Serpa" cheese whey nanofiltration for effluent minimization and by-products recovery.

Second cheese whey (SCW) is a by-product of cheese and curd cheese production that is usually not recovered and therefore substantially contributes to the negative environmental impact of the cheese manufacture plants. Membrane technology, namely nanofiltration (NF), is used in this work for the recovery of SCW organic nutrients, resulting from "Serpa" cheese and curd production. The SCW is processed by NF to recover a rich lactose fraction in the concentrate and a process water with a high salt content in the permeate. The permeation experiments were carried out in a plate & frame NF unit, where two NF membranes (NFT50 and HR-95-PP) were characterized and tested. The NF permeation experiments were performed accordingly with two different operation modes: total recirculation and concentration. In order to select the best membrane and operating pressure for the SCW fractionation, total recirculation experiments were carried out. The NF modeling was also performed, in terms of permeate fluxes and rejection coefficients using the resistances-in-series model and the solution-diffusion model, respectively. After the membrane selection, the concentration experiments showed that the selected membrane (NFT50) at 3.0MPa allows a water recovery of approximately 80%, concentrating the SCW nutrients approximately 5 times. Therefore, the NF operation can successfully reduce the wastewater organic load and simultaneously contributes to the valorization of the cheese and curd cheese manufacture by-products.

Cheese↗
PubMed2000

Effects of Lactobacillus strains on the ripening and organoleptic characteristics of Arzúa-Ulloa cheese.

Seven batches of Arzúa-Ulloa, a short-ripened soft cow's milk cheese produced in Galicia (NW Spain), were prepared from pasteurized milk. Two control batches of cheese (CB) were made with an acid-aromatic starter containing Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. lactis var. diacetylactis, isolated from raw-milk Arzúa-Ulloa cheeses. Five batches of cheese (LB) were made with the acid-aromatic starter plus one of five strains of mesophilic homofermentative Lactobacillus spp.: four of them isolated from raw-milk Arzúa-Ulloa cheese (characterized in previous works) and the remaining was a commercial Lactobacillus strain. Higher counts of mesophilic viable bacteria, lactic acid bacteria and citrate-fermenting bacteria were found on days 1 or 15 of ripening, while higher counts of lactobacilli were found on day 30 of ripening. On day 1 of ripening the highest diacetyl-acetoin content was noted in the CB, but after day 15 the diacetyl-acetoin content was similar or higher in three of the five LB. The mean degradation of beta-casein in CB was higher than in LB, while the degradation of alpha(s1)-casein was higher in LB. The mean contents of nitrogen-soluble fractions were slightly higher in the LB than in the CB. Volatile free fatty acid (VFFA) contents were, in general, greater in LB than in CB and maximum amounts were determined on day 15 of maturation. Sensorial analysis indicated a more acid taste was in LB, while bitter and astringent tastes were more intense in CB. A positive correlation was found between beta-casein degradation and bitter taste. Yogurt and butter aromas were more intense in CB and in two of the five LB. Firmness was lower in LB and a negative correlation was found between this parameter and alpha(s1)-casein degradation. Crumbliness showed a positive correlation with beta-casein degradation. The use of the Lactobacillus strains assessed in this study is recommended for Arzúa-Ulloa cheese manufacture, in order to enhance the desirable characteristics of this cheese variety, i.e., a soft texture due to alpha(s1)-casein proteolysis but without the bitter taste due to beta-casein degradation and a spicy and slightly rancid aroma and taste.

Acetoin↗
PubMed2001

Nisin-producing organisms during traditional 'Fior di latte' cheese-making monitored by multiplex-PCR and PFGE analyses.

In this work we studied using different molecular methods the population dynamics of nisin-producing organisms and the persistence of such organisms within a complex ecosystem, 'Fior di latte' cheese, a traditional high-moisture pasta filata cheese. Using the primers targeting the eubacterial 16S-23S rRNA spacer region, together with those amplifying the nisA or nisZ gene, we were able to provide a rapid species identification of the isolates. Inhibitors of Lactococcus lactis subsp. lactis DSM 20481T used as indicator occurred during the whole process of cheese manufacture as a significant part of lactic microflora; however, only 12 among 109 isolates of bacteriocin producers were nisin producers. Amplification of the nisA or nisZ gene, using DNA extracted directly from dairy samples as templates, showed that the nisin structural gene was detected during cheese-making from milk samples up to the end of curd ripening but not in the final cheese. In order to monitor nisin-producing strains during cheese manufacturing, the 12 Lactococcus lactis nis+ strains were analysed by low frequency restriction fragment and PFGE. Nine isolates among the 12 nisin-producers exhibited an unique and distinct DNA banding pattern and are considered to be genetically diverse. The other three isolates from curd after ripening showed the same restriction pattern and could be the same strain. In fact, it was also isolated 2 months after the first analysis of cheese-making of 'Fior di latte'.

Bacteriocins↗
PubMed2001

Yeasts from Water Buffalo Mozzarella, a traditional cheese of the Mediterranean area.

Countries of the Mediterranean area are characterized by production of artisanal cheeses, obtained from goat, sheep, cow and buffalo raw milk. The numbers and species of yeasts in the different cheeses are variable, but some species are more frequently detected than others. Kluyveromyces marxianus, K. lactis with their anamorph, Candida kefir, Debaryomyces hansenii and C. famata, C. colliculosa and C. catenulata are dominant species in several cheeses. However, Saccharomyces cerevisiae is often detected in pasta filata cheeses, such as Water Buffalo Mozzarella (WBM) or Cacio Cavallo Podolico. Recently, a comprehensive study of yeasts isolated from Mozzarella cheese produced in Basilicata (Southern Italy) has been carried out. The study has focused on lactose and/or galactose fermenting species (Kluyveromyces and Saccharomyces) to evaluate their role on the functional and sensory properties of the product. End products in milk were evaluated and the biodiversity in terms of production of sulphur dioxide, higher alcohols, ethyl acetate, and acetaldehyde was studied. In particular, S. cerevisiae strains from Water Buffalo Mozzarella cheese, compared to strains isolated from different habitats, such as wine, exhibited considerable difference in the production of some volatile compounds. The diversity observed could be related to the particular microhabitat of S. cerevisiae occurring in whey cheese of water buffalo milk.

Animals↗
PubMed2003

Applicability of a bacteriocin-producing Enterococcus faecium as a co-culture in Cheddar cheese manufacture.

Two strains, Enterococcus faecium RZS C5 and E. faecium DPC 1146, produce listericidal bacteriocins, so-called enterocins. E. faecium RZS C5 was studied during batch fermentation in both a complex medium (MRS) and in milk to understand the influence of environmental factors, characteristic for milk and cheese, on both growth and bacteriocin production. Fermentation conditions were chosen in view of the applicability of in situ enterocin production during Cheddar cheese production. Enterocin production by E. faecium RZS C5 in MRS started in the early logarithmic growth phase, and enterocin activity decreased during the stationary phase. The effect of pH on enterocin production and decrease of activity was as intense as the effect on bacterial growth. Higher enterocin production took place at pH 5.5 compared with pH 6.5. The use of lactose instead of glucose increased the production of enterocin, and at higher lactose concentration, production increased more and loss of activity decreased. The production in skimmed milk compared to MRS was lower and was detected mainly in the stationary phase. When casein hydrolysate was added to the milk, enterocin production was higher and started earlier, indicating the importance of an additional nitrogen source for growth of E. faecium in milk. For co-cultures of E. faecium RZS C5 with the starters used during Cheddar cheese manufacture, no enterocin activity was detected during the milk fermentation. Furthermore, the applicability of E. faecium RZS C5 and E. faecium DPC 1146 strains was tested in Cheddar cheese manufacture on pilot scale. Enterocin production took place from the beginning of the cheese manufacturing and was stable during the whole ripening phase of the cheese. This indicates that both an early and late contamination of the milk or cheese can be combated with a stable, in situ enterocin production. The use of such a co-culture is an additional safety provision beyond good manufacturing practices.

Bacteriocins↗
PubMed2003

Early lysis of Lactobacillus helveticus CNRZ 303 in Swiss cheese is not prophage-related.

Lactobacillus helveticus is mainly used as starter in Swiss-type cheeses. Often, lysogenic strains are eliminated because of the risk of early lysis and acidification failure due to phage expression. On the other hand, L. helveticus lysis was shown to positively influence cheese proteolysis during ripening. In order to better assess the relationship between lysis and lysogeny, a prophage-cured derivative of L. helveticus CNRZ 303 was isolated (LH 303-G11) and relysogenised (LH 303-G11R), as demonstrated by hybridisation using the whole phage DNA as probe. The growth, lysis in buffered solutions and lytic activities in zymogram using either Micrococcus luteus or L. helveticus as substrate were identical between the mother strain and its cured derivatives. Only morphological differences were observed by scanning electron microscopy: the cells of the cured derivative were shorter in length. The mother strain and its cured and relysogenised derivatives were assayed in triplicate in experimental Swiss cheeses (scale 1:100). No differences were noted during the cheese making: the three strains exhibited identical kinetics of acidification, leading to similar cheeses at day 1 in terms of gross composition and pH. Phages were detected only in the cheeses made with the mother strain and the relysogenised derivative. The lysis of L. helveticus, estimated by viability decrease and release of the intracellular marker D-lactate deshydrogenase, started early before brining and continued during the cold room ripening. No obvious differences of lysis extent were observed. These results demonstrated for the first time that, in the case of LH 303, the extensive lysis observed in cheese is mainly due to autolysin activity and not to prophage induction.

Bacteriolysis↗
PubMed2002

Quantitative microbial risk assessment exemplified by Staphylococcus aureus in unripened cheese made from raw milk.

This paper discusses some of the developments and problems in the field of quantitative microbial risk assessment, especially exposure assessment and probabilistic risk assessment models. To illustrate some of the topics, an initial risk assessment was presented, in which predictive microbiology and survey data were combined with probabilistic modelling to simulate the level of Staphylococcus aureus in unripened cheese made from raw milk at the time of consumption. Due to limited data and absence of dose-response models, a complete risk assessment was not possible. Instead, the final level of bacteria was used as a proxy for the potential enterotoxin level, and thus the potential for causing illness. The assessment endpoint selected for evaluation was the probability that a cheese contained at least 6 log cfu S. aureus g(-1) at the time of consumption; the probability of an unsatisfactory cheese, P(uc). The initial level of S. aureus, followed by storage temperature had the largest influence on P(uc) at the two pH-values investigated. P(uc) decreased with decreasing pH and was up to a factor of 30 lower in low pH cheeses due to a slower growth rate. Of the model assumptions examined, i.e. the proportion of enterotoxigenic strains, the level of S. aureus in non-detect cheeses, the temperature limit for toxin production, and the magnitude and variability of the threshold for an unsatisfactory cheese, it was the latter that had the greatest impact on P(uc). The uncertainty introduced by this assumption was in most cases less than a factor of 36, the same order of magnitude as the maximum variability due to pH. Several data gaps were identified and suggestions were made to improve the initial risk assessment, which is valid only to the extent that the limited data reflected the true conditions and that the assumptions made were valid. Despite the limitations, a quantitative approach was useful to gain insights and to evaluate several factors that influence the potential risk and to make some inferences with relevance to risk management. For instance, the possible effect of using starter cultures in the cheese making process to improve the safety of these products.

Animals↗
PubMed2003

The use of AFLP to relate cheese-contaminating Penicillium strains to specific points in the production plants.

Amplified fragment length polymorphism (AFLP) analysis was performed on isolates of Penicillium commune and Penicillium palitans originating from cheese and indoor environment in four cheese factories. The AFLP method was found to be a useful tool for identification of P. commune and P. palitans on, as well as below, species level. However, AFLP in combination with M13 fingerprinting described in a previous paper provided better resolution at the intraspecific level than either of the methods alone. Specific P. commune and P. palitans strains were found in the same factories over a period of more than a year and showed that the cheese factories have contaminating strains that are well established. The majority of the P. commune and P. palitans strains were found only within a single factory, but several were found in different cheese factories. The combined fingerprinting data could relate strains isolated from cheese to specific points in the production plants. Several of cheese-contaminating Penicillium strains could be related to air in the wrapping room, which must be considered to be a critical point for contamination of cheese.

Cheese↗
PubMed2000

Influence of milk centrifugation, brining and ripening conditions in preventing gas formation by Clostridium spp. in Gouda cheese.

This study examined milk centrifugation, increased salt concentration, and low ripening temperature as potential strategies to prevent late blowing caused by gas-forming Clostridium spp. in Gouda cheese. The survival of clostridia spores in cheese brine and their ability to enter Gouda cheese during brining was also evaluated. Centrifugation (3000 x g for 30 s) of contaminated milk resulted in > 60% spore reduction, with increased spore reduction at greater centrifugal forces. Low levels of C. tyrobutyricum and C. sporogenes spores survived in saturated (23%, w/v) brine with 2% (v/v) added whey at 15 degrees C for 63 days, while C. beijerinckii and C. butyricum spores were not detectable on days 4 and 35, respectively. Spores of C. tyrobutyricum in brine infiltrated Gouda cheese during 2 h of brining at 13 degrees C resulted in production of small gas holes during ripening. In Gouda cheese slurry stored at 13 degrees C, three C. tyrobutyricum strains plus one of three C. sporogenes strains germinated in the slurry with no added salt. Of three C. tyrobutyricum strains stored at 13 degrees C in slurries with higher water-phase salt concentrations of 2.4 and 3.6%, two strains and one strain germinated, respectively. No germination of spores was detected in any cheese slurry stored at 5 or 8 degrees C. Milk centrifugation, increased percent water-phase salt, absence of spores in brine, and decreased ripening temperature are all potentially important measures against gas production by Clostridium spp. in Gouda cheese.

Animals↗
PubMed2004

90Sr, 238U, 234U, 137Cs, 40K and 239/240Pu in Emmental type cheese produced in different regions of Western Europe.

A method is presented for the determination of (90)Sr and uranium in Emmental type cheese collected in dairy plants from different European countries. Results display a significant correlation (r = 0.708 Student t-test = 6.02) between the (90)Sr content of the cheese and the altitude of grazing. The highest (90)Sr activity is 1.13 Bq kg(-1) of cheese and the lowest is 0.29 Bq kg(-1). Uranium activity is very low with a highest (238)U value of 27 mBq kg(-1). In addition, (234)U/(238)U ratio shows a large enrichment in (234)U for every location. Without any significant indication of the geographic origin of the cheese, this enrichment is believed to be due to the geological features of the pasture, soil and underground water. These results tend to prove that the contamination of milk by uranium originates principally from the water that the cows drink instead of the forage. This finding may have a great importance in models dealing with dairy food contamination by radionuclides following a nuclear accident. Also, the (90)Sr content and to a lesser extent the (234)U/(238)U ratio could be used to trace the authenticity of the origin of the cheese. (137)Cs activity is lower than the detection limit of 0.1 Bq kg(-1) in all the samples collected (n = 20). Based on natural (40)K activity in cheese (15-21 Bq kg(-1)), the decontamination factor for the alkaline cations from milk to cheese is about 20. Plutonium activity stays below the detection limit of 0.3 mBq kg(-1).

Animals↗
PubMed2001

Anaerobic biotransformation and methane generation potential of cheese whey in batch and UASB reactors.

Anaerobic treatability and methane generation potential of cheese whey were determined in batch reactors. Furthermore, the effect of nutrient and trace metal supplementation on the batch anaerobic treatment, and the high-rate anaerobic treatability of cheese whey in upflow anaerobic sludge blanket (UASB) reactors were investigated. To this purpose biochemical methane potential experiments were conducted and single- and two-stage UASB reactors with granular cultures were operated. In UASB experiments significance of process staging, operational parameters such as hydraulic retention time (HRT), influent chemical oxygen demand (COD) concentration and loading rate were also investigated. The results revealed that nutrient and trace metal supplementation is vital for the anaerobic treatment of cheese whey; the anaerobic methane generation for the cheese whey studied was found to be 424 ml CH4/g COD (23.4 1 CH4/l cheese whey); undiluted cheese whey could be treated anaerobically at relatively short HRT values (2.06-4.95 days) without any significant stability problems; HRT values as low as 2-3 days can be used for the anaerobic treatment of cheese whey, with a COD removal efficiency of 95-97% at influent COD concentration of 42 700 +/- 141-55 100 +/- 283 mg/l.

Bacteria, Anaerobic↗
PubMed2000

Autolysis and related proteolysis in Swiss cheese for two Lactobacillus helveticus strains.

Intracellular peptidases of Lactobacillus helveticus may play a major role in the proteolysis of Swiss cheeses, provided that they are released through bacterial lysis. Experimental Swiss cheeses were manufactured on a small scale from thermized and microfiltered milk using as starters (in addition to Streptococcus thermophilus and Propionibacterium freudenreichii) one of two Lb. helveticus strains, ITGLH1 and ITGLH77, which undergo lysis to different extents in vitro. All the cheeses were biochemically identical after pressing. The viability of Lb. helveticus ITGLH1 and ITGLH77 decreased to a similar extent (96-98%) while in the cold room, but the concomitant release of intracellular lactate dehydrogenase in cheeses made with strain ITGLH1 was 5-7-fold that in cheeses made with ITGLH77. Protein profiles and immunoblot detection of the dipeptidase PepD confirmed a greater degree of lysis of the ITGLH1 strain. Free active peptidases were detected in aqueous extracts of cheese for both strains, and proteolysis occurred principally in the warm room. Reversed-phase HPLC revealed a more extensive peptide hydrolysis for ITGLH1, which was confirmed by the greater release of free NH2 groups (+33%) and free amino acids (+75%) compared with ITGLH77. As the intracellular peptidase activities of ITGLH1 and ITGLH77 have previously been shown to be similar, our results indicated that the extent of lysis of Lb. helveticus could have a direct impact on the degree of proteolysis in Swiss cheeses.

Autolysis↗