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PubMed2002

Antibacterial activities of nisin Z encapsulated in liposomes or produced in situ by mixed culture during cheddar cheese ripening.

This study investigated both the activity of nisin Z, either encapsulated in liposomes or produced in situ by a mixed starter, against Listeria innocua, Lactococcus spp., and Lactobacillus casei subsp. casei and the distribution of nisin Z in a Cheddar cheese matrix. Nisin Z molecules were visualized using gold-labeled anti-nisin Z monoclonal antibodies and transmission electron microscopy (immune-TEM). Experimental Cheddar cheeses were made using a nisinogenic mixed starter culture, containing Lactococcus lactis subsp. lactis biovar diacetylactis UL 719 as the nisin producer and two nisin-tolerant lactococcal strains and L. casei subsp. casei as secondary flora, and ripened at 7 degrees C for 6 months. In some trials, L. innocua was added to cheese milk at 10(5) to 10(6) CFU/ml. In 6-month-old cheeses, 90% of the initial activity of encapsulated nisin (280 +/- 14 IU/g) was recovered, in contrast to only 12% for initial nisin activity produced in situ by the nisinogenic starter (300 +/- 15 IU/g). During ripening, immune-TEM observations showed that encapsulated nisin was located mainly at the fat/casein interface and/or embedded in whey pockets while nisin produced by biovar diacetylactis UL 719 was uniformly distributed in the fresh cheese matrix but concentrated in the fat area as the cheeses aged. Cell membrane in lactococci appeared to be the main nisin target, while in L. casei subsp. casei and L. innocua, nisin was more commonly observed in the cytoplasm. Cell wall disruption and digestion and lysis vesicle formation were common observations among strains exposed to nisin. Immune-TEM observations suggest several modes of action for nisin Z, which may be genus and/or species specific and may include intracellular target-specific activity. It was concluded that nisin-containing liposomes can provide a powerful tool to improve nisin stability and availability in the cheese matrix.

Anti-Bacterial Agents↗
PubMed2002

Inhibition of Listeria innocua in cheddar cheese by addition of nisin Z in liposomes or by in situ production in mixed culture.

The effect of addition of purified nisin Z in liposomes to cheese milk and of in situ production of nisin Z by Lactococcus lactis subsp. lactis biovar diacetylactis UL719 in the mixed starter on the inhibition of Listeria innocua in cheddar cheese was evaluated during 6 months of ripening. A cheese mixed starter culture containing Lactococcus lactis subsp. lactis biovar diacetylactis UL719 was selected for high-level nisin Z and acid production. Experimental cheddar cheeses were produced on a pilot scale, using the selected starter culture, from milk with added L. innocua (10(5) to 10(6) CFU/ml). Liposomes with purified nisin Z were prepared from proliposome H and added to cheese milk prior to renneting to give a final concentration of 300 IU/g of cheese. The nisin Z-producing strain and nisin Z-containing liposomes did not significantly affect cheese production and gross chemical composition of the cheeses. Immediately after cheese production, 3- and 1.5-log-unit reductions in viable counts of L. innocua were obtained in cheeses with encapsulated nisin and the nisinogenic starter, respectively. After 6 months, cheeses made with encapsulated nisin contained less than 10 CFU of L. innocua per g and 90% of the initial nisin activity, compared with 10(4) CFU/g and only 12% of initial activity in cheeses made with the nisinogenic starter. This study showed that encapsulation of nisin Z in liposomes can provide a powerful tool to improve nisin stability and inhibitory action in the cheese matrix while protecting the cheese starter from the detrimental action of nisin during cheese production.

Cheese↗
PubMed2006

Identification of a nisI promoter within the nisABCTIP operon that may enable establishment of nisin immunity prior to induction of the operon via signal transduction.

Certain strains of Lactococcus lactis produce the broad-spectrum bacteriocin nisin, which belongs to the lantibiotic class of antimicrobial peptides. The genes encoding nisin are organized in three contiguous operons: nisABTCIP, encoding production and immunity (nisI); nisRK, encoding regulation; and nisFEG, also involved in immunity. Transcription of nisABTCIP and nisFEG requires autoinduction by external nisin via signal transducing by NisRK. This organization poses the intriguing question of how sufficient immunity (NisI) can be expressed when the nisin cluster enters a new cell, before it encounters external nisin. In this study, Northern analysis in both Lactococcus and Enterococcus backgrounds revealed that nisI mRNA was present under conditions when no nisA transcription was occurring, suggesting an internal promoter within the operon. The nisA transcript was significantly more stable than nisI, further substantiating this. Reverse transcriptase PCR analysis revealed that the transcription initiated just upstream from nisI. Fusing this region to a lacZ gene in a promoter probe vector demonstrated that a promoter was present. The transcription start site (TSS) of the nisI promoter was mapped at bp 123 upstream of the nisI translation start codon. Ordered 5' deletions revealed that transcription activation depended on sequences located up to bp -234 from the TSS. The presence of poly(A) tracts and computerized predictions for this region suggested that a high degree of curvature may be required for transcription initiation. The existence of this nisI promoter is likely an evolutionary adaptation of the nisin gene cluster to enable its successful establishment in other cells following horizontal transfer.

Bacterial Proteins↗
PubMed1994

Characterization of the lactococcal temperate phage TP901-1 and its site-specific integration.

The temperate lactococcal phage TP901-1, induced by UV light from Lactococcus lactis subsp. cremoris 901-1, was characterized. The restriction map was found to be circular, and the packaging of TP901-1 DNA was concluded to occur by a headful mechanism. The pac region was localized on the 38.4-kb phage genome. TP901-1 belongs to the class of P335 phages (V. Braun, S. Hertwig, H. Neve, A. Geis, and M. Teuber, J. Gen. Microbiol. 135:2551-2560, 1989). Evidence is presented that the phages TP936-1 (V. Braun, S. Hertwig, H. Neve, A. Geis, and M. Teuber, J. Gen. Microbiol. 135:2551-2560, 1989) and C3-T1 (A. W. Jarvis, V. R. Parker, and M. B. Bianchin, Can. J. Microbiol. 38:398-404, 1992) are very closely related to or are identical to TP901-1. The lytically propagated TP901-1 phages were able to lysogenize both indicator strains Lactococcus cremoris 3107 and Wg2. Lysogenization resulted in site-specific integration of the phage genome into the bacterial chromosome. Only one chromosomal attB site was found in 20 independent lysogens. The attP region of TP901-1 and the attL and attR regions were cloned and sequenced. The results showed a core region of only 5 bp, in which the recombination occurs, followed after a 1-bp mismatch by a 7-bp identical region, TCAAT(T/C)AAGGTAA. This result was further verified by sequencing of the attB region obtained by PCR. An integration vector was constructed with the 6.5-kb EcoRI fragment from TP901-1 containing attP. This vector also functions in the plasmid-free strains, MG1363 and LM0230 with only one specific attB site, strongly indicating a more general use of the TP901-1-based integration vector in lactococci.

Bacteriophages↗
PubMed1995

Distribution of proteins similar to IIIManH and IIIManL of the Streptococcus salivarius phosphoenolpyruvate:mannose-glucose phosphotransferase system among oral and nonoral bacteria.

In Streptococcus salivarius, the phosphoenolpyruvate (PEP):mannose-glucose phosphotransferase system, which concomitantly transports and phosphorylates mannose, glucose, fructose, and 2-deoxyglucose, is composed of the general energy-coupling proteins EI and HPr, the specific membrane-bound IIIMan, and two forms of a protein called IIIMan, with molecular weights of 38,900 (IIIManH) and 35,200 (IIIManL), that are found in the cytoplasm as well as associated with the membrane. Several lines of evidence suggest that IIIManH and/or IIIManL are involved in the control of sugar metabolism. To determine whether other bacteria possess these proteins, we tested for their presence in 28 oral streptococcus strains, 3 nonoral streptococcus strains, 2 lactococcus strains, 2 enterococcus strains, 2 bacillus strains, 1 lactobacillus strain, Staphylococcus aureus, and Escherichia coli. Three approaches were used to determine whether the IIIMan proteins were present in these bacteria: (i) Western blot (immunoblot) analysis of cytoplasmic and membrane proteins, using anti-IIIManH and anti-IIIManH rabbit polyclonal antibodies; (ii) analysis of PEP-dependent phosphoproteins by polyacrylamide gel electrophoresis; and (iii) inhibition by anti-IIIMan antibodies of the PEP-dependent phosphorylation of 2-deoxyglucose (a mannose analog) by crude cellular extracts. Only the species S. salivarius and Streptococcus vestibularis possessed the two forms of IIIMan. Fifteen other streptococcal species possessed one protein with a molecular weight between 35,200 and 38,900 that cross-reacted with both antibodies. In the case of 9 species, a protein possessing the same electrophoretic mobility was phosphorylated at the expense of PEP. No such phosphoprotein, however, could be detected in the other six species. A III(Man)-like protein with a molecular weight of 35,500 was also detected in Lactobacillus casei by Western blot experiments as well as by PEP-dependent phosphoprotein analysis, and a protein with a molecular weight of 38,900 that cross-reacted with anti-III(Man) antibodies was detected in Lactococcus lactis. In several cases, the involvement of these putative III(Man) proteins in the PEP-dependent phosphorylation of 2-deoxyglucose was substantiated by the inhibition of phosphorylation activity of anti-III(Man) antibodies. No proteins cross-reacting with anti-III(Man) antibodies were detected in enterococci, bacilli, and E. coli. In S. aureus, a membrane protein with a molecular weight of 50,000 reacted strongly with the antibodies. This protein, however, was not phosphorylated at the expense of PEP.

Bacillus↗
PubMed2003

Survival of lactococci during passage through mouse digestive tract.

One of the important properties of probiotics is the ability to survive in the intestine. There have been few studies on the probiotic property of lactococci, since they are formally not considered to be natural inhabitants of the intestine. To evaluate lactococci as probiotic bacteria, we investigated their ability to survive during gastric transit by in vitro and in vivo tests. When exposed to an in vitro simulated gastrointestinal environment, such as low pH and bile, only Lactococcus lactis subsp. lactis bv. diacetylactis N7 showed a moderate survival rate among the four strains tested. The tested strains were orally administered to mice, and intestinal passage of the ingested strains was monitored by two methods: antibiotics and PCR. Viable cells of strain N7 were recovered from feces within 24-48 h after administration but not at 72 h. Lactococcus lactis subsp. cremoris ATCC 19257, which had a poor survival rate in vitro test, was also detected at 12 h but not at 24 h. These results indicate that lactococci can reach the mouse intestine alive, but not colonize it. If administered daily, viable strain N7 may exist continuously in the intestine. The effect of strain N7 on intestinal microbial balance and on animal health will be the subject of a further study.

Animals↗
PubMed2001

Isolation, characterization, and influence of native, nonstarter lactic acid bacteria on Cheddar cheese quality.

To determine whether adventitious nonstarter lactic acid bacteria (NSLAB) might affect cheese flavor and quality, we studied a population of NSLAB present in 30 premium quality Cheddar cheeses (3-mo ripened) produced at a commercial facility in the United States. DNA fingerprinting analysis with a sensitive strategy for arbitrary priming polymerase chain reaction showed that 75 isolates corresponded to at least 18 distinct nonstarter organisms. According to ribotype database comparisons of representatives from the 18 groups, 9 matched Lactobacillus (closest to paracasei species), 8 matched Streptococcus thermophilus, and 1 matched to a Lactococcus species. This finding indicated that among the 75 NSLAB isolates, Lactobacillus made up 64%, S. thermophilus 32%, and Lactococcus 4%. Isolates representing 11 NSLAB groups were characterized for protease, peptidase, and diacetyl production. Based on this phenotypic analysis, two Lactobacillus isolates were evaluated as adjuncts in Cheddar cheese. All of the NSLAB identified from the adjunct cheese at 3 mo by DNA fingerprinting consisted of the adjunct lactobacilli, showing that the adjunct strains predominated throughout the early stages of ripening. The impact of adjunct lactobacilli was evident after 6 mo when free amino acids significantly increased and sensory scores improved in adjunct cheese as compared with a control cheese. The largest impact was found in adjunct cheese containing a blend of both lactobacilli strains. These results show that certain adventitious NSLAB positively contribute to flavor development.

Amino Acids↗
PubMed2001

Influence of adjunct cultures on ripening of reduced fat Edam cheeses.

The influence of four adjunct cultures [Brevibacterium linens (BL2), Lactococcus lactis ssp. diacetylactis, Lactobacillus helveticus (LH212), and Lactobacillus reuteri (ATCC 23272)] on chemical and sensory characteristics of reduced fat Edam cheese was studied. The aminopeptidase activity of Lactococcus lactis ssp. diacetylactis was higher than that of Lactobacillus helveticus, Lactobacillus reuteri, and Brevibacterium linens, respectively. Mean percent fat and moisture contents of reduced fat cheese were 20.85 +/- 0.69 and 42.95 +/- 0.43, respectively. Percentage of fat and moisture of full fat control cheese were 30.06 +/- 0.78 and 39.11 +/- 0.60. Titratable acidity increased in all cheese with aging while pH initially decreased but increased in cheese after 6 mo aging at 7 degrees C. Lactic acid bacteria counts were on average one log higher for reduced fat cheeses than for full fat control cheese and counts decreasing with aging. Free amino acids (FAA) in cheeses increased with aging, and were higher in reduced fat cheeses than in the full fat control cheese. Reduced fat cheeses containing L. helveticus exhibited the highest FAA content. Descriptive sensory panelists (n = 9) did not detect differences among cheeses after 3 and 6 mo ripening, but aged/developed flavors (fruity, nutty, brothy, sulfur, free fatty acid) and sweetness increased between 3 and 6 mo. Expert panelists (n = 6) detected differences in texture quality among the cheeses. Reduced fat control cheeses and reduced fat cheeses with L. helveticus and L. reuteri received the highest texture quality scores. Addition of L. helveticus and Lc. lactis ssp. diacetylactis, as adjunct cultures to reduced fat Edam cheeses increased proteolysis, while the addition of L. helveticus and L. reuteri increased texture quality of cheeses.

Amino Acids↗
PubMed2004

Factors affecting calcium lactate and liquid expulsion defects in Cheddar cheese.

This paper summarizes the results of 2 studies designed to investigate the influence of several manufacturing and curing treatments on the appearance of Cheddar cheese defects. Specifically, 2 defects, calcium lactate crystal formation and the expulsion of free liquid (weeping) were monitored in Cheddar cheese. Both studies were conducted at a commercial cheese manufacturing facility that produces Cheddar in 18.14-kg (40-lb) blocks. In the first study we monitored cheese calcium, both total and soluble during manufacture and early curing. In the second study we measured cheese pH from 3 d through 8 mo, as well as some factors that are influenced by cheese pH. Early cheese pH (3 d to 7 d) patterns were used to select vats of cheese for retail packaging. Mild Cheddar packaged at 30 d postmanufacture and sharp Cheddar packaged at 8 mo postmanufacture from the same vats were monitored for the incidence and severity of the defects. Our results indicated that factors measured in early stages of manufacture and curing (less than 7 d) such as cheese pH at mill, lactic acid concentration, nonprotein nitrogen, and calcium (total and soluble) in cheese did not correlate with the appearance of either calcium lactate or expulsion of free liquid in packaged cheeses. Factors including pH, lactic acid concentrations, and soluble calcium measured during curing (greater than 7 d) of cheese were found to be statistically significant in the development of defects and appeared to be associated with use of specific starter culture groups. In the study, 5 different starter culture groups, each consisting of a 4-strain blend of Lactococcus lactis ssp. cremoris and Lactococcus lactis ssp. lactis, were used to manufacture the cheeses. Cheese manufactured with one particular culture group showed no incidence of calcium lactate crystal formation or weeping during curing and shelf-life of cheeses in this study. This starter group also generated the least amount of pH change in cheese during the first month of curing. From these results we conclude that starter culture group, more than any other factor measured, played an important role in the development of calcium lactate and liquid expulsion defects in Cheddar cheese. Starter culture group appeared to strongly influence cheese pH, lactic acid, and soluble calcium concentrations during curing and storage.

Calcium↗
PubMed2005

Use of an alpha-galactosidase gene as a food-grade selection marker for Streptococcus thermophilus.

The alpha-galactosidase gene (aga) of Lactococcus raffinolactis ATCC 43920 was previously shown to be an efficient food-grade selection marker in Lactococcus lactis and Pediococcus acidilactici but not in Streptococcus thermophilus. In this study, we demonstrated that the alpha-galactosidase of L. raffinolactis is thermolabile and inoperative at 42 degrees C, the optimal growth temperature of S. thermophilus. An in vitro assay indicated that the activity of this alpha-galactosidase at 42 degrees C was only 3% of that at 30 degrees C, whereas the enzyme retained 23% of its activity at 37 degrees C. Transformation of Strep. thermophilus RD733 with the shuttle-vector pNZ123 bearing the aga gene of L. raffinolactis (pRAF301) generated transformants that were stable and able to grow on melibiose and raffinose at 37 degrees C or below. The transformed cells possessed 6-fold more alpha-galactosidase activity after growth on melibiose than cells grown on lactose. Slot-blot analyses of aga mRNA indicated that repression by lactose occurred at the transcriptional level. The presence of pRAF301 did not interfere with the lactic acid production when the transformed cells of Strep. thermophilus were grown at the optimal temperature in milk. Using the recombinant plasmid pRAF301, which carries a chloramphenicol resistance gene in addition to aga, we showed that both markers were equally efficient at differentiating transformed from nontransformed cells. The aga gene of L. raffinolactis can be used as a highly efficient selection marker in Strep. thermophilus.

Dairy Products↗
PubMed2005

Influence of starters on chemical, biochemical, and sensory changes in Turkish White-brined cheese during ripening.

Turkish White-brined cheese was manufactured using Lactococcus strains (Lactococcus lactis ssp. lactis NCDO763 plus L. lactis ssp. cremoris SK11 and L. lactis ssp. lactis UC317 plus L. lactis ssp. cremoris HP) or without a starter culture, and ripened for 90 d. It was found that the use of starters significantly influenced the physical, chemical, biochemical, and sensory properties of the cheeses. Chemical composition, pH, and sensory properties of cheeses made with starter were not affected by the different starter bacteria. The levels of soluble nitrogen fractions and urea-PAGE of the pH 4.6-insoluble fractions were found to be significantly different at various stages of ripening. Urea-PAGE patterns of the pH 4.6-insoluble fractions of the cheeses showed that considerable degradation of alpha(s1)-casein occurred and that beta-casein was more resistant to hydrolysis. The use of a starter culture significantly influenced the levels of 12% trichloroacetic acid-soluble nitrogen, 5% phosphotungstic acid-soluble nitrogen, free amino acids, total free fatty acids, and the peptide profiles (reverse phase-HPLC) of 70% (vol/vol) ethanol-soluble and insoluble fractions of the pH 4.6-soluble fraction of the cheeses. The levels of peptides in the cheeses increased during the ripening period. Principal component and hierarchical cluster analyses of electrophoretic and chromatographic results indicated that the cheeses were significantly different in terms of their peptide profiles and they were grouped based on the use and type of starter and stage of ripening. Levels of free amino acid in the cheeses differed; Leu, Glu, Phe, Lys, and Val were the most abundant amino acids. Nitrogen fractions, total free amino acids, total free fatty acids, and the levels of peptides resolved by reverse phase-HPLC increased during ripening. No significant differences were found between the sensory properties of cheeses made using a starter, but the cheese made without starter received lower scores than the cheeses made using a starter. It was found that the cheese made with strains NCDO763 plus SK11 had the best quality during ripening. It was concluded that the use of different starter bacteria caused significant differences in the quality of the cheese, and that each starter culture contributed to proteolysis to a different degree.

Amino Acids↗
PubMed2006

Monitoring the chemical and textural changes during ripening of Iranian White cheese made with different concentrations of starter.

The effect of the concentration of starter inoculated to milk on the composition, free tyrosine-tryptophan content, microstructure, opacity, and fracture stress of Iranian White cheese (IWC) was studied during 50 d of ripening in brine. Three treatments of cheese were made using 1-fold (IWC1S), 2-fold (IWC2S), and 4-fold (IWC4S) concentrations of a direct-to-vat mesophilic mixed culture containing Lactococcus lactis ssp. cremoris and Lactococcus lactis ssp. lactis as starter. As ripening progressed, moisture and protein contents of the treatments continuously decreased, whereas their total ash, salt, and salt in moisture contents increased. Fat content and pH of cheeses remained stable during ripening. The pH of cheese milk at the time of renneting, which decreased by increasing the concentration of starter (6.57, 6.49, and 6.29 for IWC1S, IWC2S, and IWC4S, respectively), significantly affected most of the chemical characteristics and opacity of cheese. Lower pH values at renneting decreased moisture and ash contents, whereas cheese protein content increased. The concentration of free tyrosine-tryptophan in curd increased at first 29 d but decreased between d 29 and 49 of aging. The changes observed in cheese whiteness followed the changes in moisture content of the treatments. As the concentration of starter inoculated to milk increased, the value of fracture stress at a given ripening time significantly decreased, leading to a less resistant body against applied stress. A similar trend was also observed for fracture strain during cheese ripening. The micrographs taken by scanning electron microscopy provided a meaningful explanation for decrease in the value of fracture stress. As the cheese ripening progressed or the concentration of starter increased, the surface area occupied by the protein fraction in cheese microstructure decreased, leading the way to lower the force-bearing component in cheese texture.

Animals↗
PubMed1991

Microbiological analysis and starter culture growth in retentates.

Pasteurized skim milk was concentrated by UF to 2-, 4-, and 5-fold. The retentates were evaluated for microbiological quality, heat treatments to inactivate microorganisms, and lactic acid bacterial starter culture activity. Aerobic mesophilic bacterial counts in raw milk decreased from an initial 1.4 x 10(6) to 3.9 x 10(2) cfu/ml after pasteurization. During UF, counts increased from 3.9 x 10(2) cfu/ml UF, counts increased from 3.9 x 10(2) cfu/ml in pasteurized milk to 1.4 x 10(3), 1.4 x 10(4), and 1.8 x 10(4) cfu/ml in 2-, 4- and 5-fold retentates, respectively. Psychrotrophic bacterial counts decreased from 9.9 x 10(5) cfu/ml in raw milk to 3.7 x 10(1) cfu/ml in pasteurized milk and gradually increased to 1.0 x 10(2), 2.5 x 10(2), and 1.4 x 10(3) cfu/ml in 2-, 4-, and 5-fold retentates, respectively. Thermophilic bacterial counts remained less than 10 cfu/ml in all samples. Skim milk and retentates inoculated with five starter cultures at 1% failed to decrease the pH below 4.6 in (2-, 4- and 5-fold). The 4- and 5-fold retentates inoculated with Lactococcus lactis spp. cremoris or Lactococcus lactis spp. lactis cultures were partially coagulated with pH greater than 5.6. In general, the pH of retentates remained higher than that of skim milk. Clotting of uninoculated samples was observed, and a spore-forming contaminant, tentatively characterized as Bacillus cereus and capable of clotting milk at a pH greater than 6, was isolated from the clotted samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2000

Behavior of Listeria monocytogenes in pasteurized milk during fermentation with lactic acid bacteria.

The behavior of Listeria monocytogenes in pasteurized milk during fermentation with starter and nonstarter lactic acid bacteria was investigated. Pasteurized milk was co-inoculated with approximately 10(4) CFU/ml of L. monocytogenes and 10(6) CFU/ml of Lactococcus lactis, Lactococcus cremoris, Lactobacillus plantarum, Lactobacillus bulgaricus, or Streptococcus thermophilus. Inoculated milks were incubated at 30 degrees C or 37 degrees C for 24 to 72 h. Listeria monocytogenes survived and also grew to some extent during incubation in the presence of all starter cultures; however, inhibition ranged from 83 to 100% based on maximum cell populations. During incubation with L. bulgaricus and L. plantarum, L. monocytogenes was completely inactivated after 20 h and 64 h of incubation at 37 degrees C and 30 degrees C, respectively. The pH of the fermenting milks declined steadily throughout the fermentation periods and was approximately 4.2 at the conclusion of the experimental period regardless both of the starter culture and pathogen combination or the temperature of incubation.

Animals↗
PubMed2002

Esterolytic and lipolytic activities of lactic acid bacteria isolated from ewe's milk and cheese.

In the present work, we report on the esterase and lipase activities of lactic acid bacteria representing the genera Lactococcus, Leuconostoc, Lactobacillus, and Enterococcus isolated from ewe's milk and cheeses. Esterase activity was studied using alpha- and beta-naphthyl derivatives of 2 to 12 carbon atoms and postelectrophoretic detection. The lactic acid bacteria evaluated had intracellular esterase activities, which preferentially degraded the alpha- and beta-naphthyl derivatives of 2 to 6 carbon atoms. By studying postelectrophoretic patterns, it was found that some strains presented more than one esterase. Lactobacillus plantarum O236 showed four enzymes that hydrolyze carboxyl ester linkages with different specificity. Lipase activity was studied in intracellular and extracellular fractions using tributyrin, tricaprylin, triolein, and milk fat as substrates. The intracellular and extracellular fractions of Leuconostoc mesenteroides O257, Lactobacillus plantarum O236, and Lactobacillus acidophilus O177 were able to hydrolyze tributyrin. L. plantarum O186, L. acidophilus O252, Enterococcus faecium O174 and O426, and Enterococcus faecalis Ov409 showed lipase activity associated with the intracellular fraction on tributyrin. Lactococcus lactis O233, L. plantarum O155, and Lactobacillus casei O190 did not hydrolyze triglycerides. Not all strains that showed esterase activity exhibited high activity on triglycerides. Esterase and lipase activities were species- and strain-specific. Wide variations in activity between strains highlight the need for selecting appropriate starters to produce enzyme-modified cheese as well as accelerated ripened cheese.

Animals↗
PubMed2002

Competition of thermally injured listeria monocytogenes with a mesophilic lactic acid starter culture in milk for various heat treatments.

Overnight tryptose broth cultures of three L monocytogenes strains were combined, centrifuged, suspended in 200 ml of tryptose phosphate broth, and heated at 56 degrees C for 20 min and at 64 degrees C for 2 min to obtain low-heat-injured (LHI) and high-heat-injured (HHI) cells, respectively, showing >99.6% injury. Flasks containing 200 ml of raw, low-heat-treated (56 degrees C for 20 min), high-heat-treated (64 degrees C for 2 min), pasteurized, and ultrahigh-temperature (UHT) milk were tempered to 31.1 degrees C and inoculated to contain 10(4) to 10(6) CFU/ml of LHI, HHI, or healthy L. monocytogenes cells and a commercial Lactococcus lactis subsp. lactis-Lactococcus lactis subsp. cremoris starter culture at levels of 0.5, 1.0, and 2.0%. Numbers of healthy and injured L. monocytogenes cells and starter organisms were determined using tryptose phosphate agar with or without 4.0% NaCl at selected intervals during 24 h of incubation at 31.1 degrees C. The presence of L. monocytogenes did not adversely affect the growth of the starter culture at any inoculation level. Overall, L. monocytogenes survived the 24-h fermentation period and grew to some extent. In starter-free controls. 76 to 81% of LHI cells and 59 to 69% of HHI cells were repaired after 8 h of incubation, with the lowest repair rates being observed for raw rather than heat-treated or pasteurized milk. Increased injury was observed for healthy L. monocytogenes cells at the 1.0 and 2.0% starter levels, with less injury seen for LHI and HHI cells. Raw and subpasteurized milk allowed less of a decrease in the percentage of injury and also showed higher numbers of injured cells than did pasteurized and UHT milks. These findings may have important implications for the survival of Listeria spp. in certain cheeses that can be prepared from raw or heat-treated milk.

Animals↗
PubMed2004

Changes in galactose and lactic acid content of sweet whey during storage.

Whey is often stored or transported for a period of time prior to processing. During this time period, galactose and lactic acid concentrations may accumulate, reducing the quality of spray-dried whey powders in regard to stickiness and agglomeration. This study surveyed industry samples of Cheddar and mozzarella cheese whey streams to determine how galactose and lactic acid concentrations changed with storage at appropriate (4 degrees C) and abuse (37.8 degrees C) temperatures. Samples stored at 4 degrees C did not exhibit significant increases in levels of lactic acid or galactose. Mozzarella whey accumulated the greatest amount of galactose and lactic acid with storage at 37.8 degrees C. Whey samples derived from cheese made from single strains of starter culture were also evaluated to determine each culture's contribution to galactose and lactic acid production. Starter cultures evaluated included Streptococcus salivarius ssp. thermophilus. Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, Lactococcus lactis ssp. cremoris, and Lactococcus lactis ssp. lactis. Whey derived from L. helveticus accumulated a significantly greater amount of lactic acid upon storage at 37.8 degrees C as compared with the other cultures. Galactose accumulation was significantly decreased in whey from L. lactis ssp. lactis stored at 37.8 degrees C in comparison with the other cultures. Results from this study indicate that proper storage conditions (4 degrees C) for whey prevent accumulation of galactose and lactic acid while the extent of accumulation during storage at 37.8 degrees C varies depending on the culture(s) used in cheese production.

Cheese↗
PubMedDate not supplied

[Effect of products of alphaS1-casein proteolysis on the activity of angiotensin-converting enzyme].

The influence of alpha s1-casein proteolysis products of cow milk on the activity of angiotensin-converting enzyme (ACE) has been investigated. The peptide fraction has been obtained after incubation of alpha s1-casein with different strains Lactococcus lactis ssp. lactis and pepsin. The peptides with low molecular weight has been obtained with the help of a gel filtration. It is shown, that such peptides received with the help of some strains of Lactococcus lactis ssp. lactis and pepsin are capable to inhibit the activity of ACE. A conclusion about possibility of appearing antihypertensive peptides as a result of proteolitic processes in milk products has been made.

Caseins↗