Effect of exopolysaccharide-producing starter cultures and post-fermentation mechanical treatment on textural properties and microstructure of low fat yoghurt

The microstructure and texture of yoghurts produced by four different exopolysaccharide (EPS)-producing starter cultures and mechanically treated post-fermentation at four levels of intensity (applied back-pressure) were studied. Two Lactobacillus delbrueckii ssp. bulgaricus (LB) strains were used in combination with two Streptococcus thermophilus (ST) strains and yoghurts were formulated by pairwise combining one LB and one ST strain. The choice of ST strain was the major determinant for the rheological properties of the yoghurts, since one of the ST strains conferred a ropy texture and resulted in yoghurts with decreased water holding capacity and an open microstructure. In addition, one of the LB strains used produced both aggregated and threadlike EPS and improved water holding. When combined with an ST strain that produced neglible amounts of EPS this LB strain resulted in yoghurt where a moderate mechanical treatment post-fermentation was able to further improve the water holding capacity.     

Isolation and identification of high viscosity-producing lactic acid bacteria from a traditional fermented milk in Xinjiang and its role in fermentation process

Forty-four lactic acid bacteria (LAB) isolated from traditional Sayram ropy fermented milk (SRFM) in southern Xinjiang of China. Further two strains were selected based on their high viscosity-producing activity. Based on the API 50 CHL strip and 16S rDNA sequence analysis, strain MB 2-1 was a Gram-positive, rod-shaped Lactobacillus helveticus and strain MB 5-1 was identified as Streptococcus thermophilus. Both the two LAB strains were grown in the milk fermentations for pure and mixed cultures and were evaluated for their growth, acidification properties, EPS production, and ability to increase the apparent viscosity of fermented milk, respectively. L. helveticus MB 2-1 displayed a high increasing in viable cell counts and the acidifying capacity in pure cultures, whereas the relatively high EPS production and viscosity-producing capacity detected in pure culture with S. thermophilus MB 5-1 as starter culture. In addition, the mixed culture of the two strains showed a higher cell growth, EPS production, and high viscosity-producing capacity at 37?°C. Values of apparent viscosity were 4.03- and 2.41-fold higher in mixed culture than for pure cultures of L. helveticus MB 2-1 and S. thermophilus MB 5-1, respectively. There was a positive correlation between the viscosity and high molecular weight EPS production with pure and mixed cultures. Our data showed two strains combination, with high viscosity-producing and acidifying capacity, can be used in mixed cultures for the manufacture of fermented milk with improved functional properties.     

Evaluation of the yield,molar mass of exopolysaccharides,and rheological properties of gels formed during fermentation of milk by Streptococcus thermophilus strains St-143 and ST-10255y

The yield and chemical structures of exopolysaccharides (EPS) produced by many strains of Streptococcus thermophilus have been characterized. However, the kinetics (or production profile) for EPS during milk fermentation is not clear. In this study, we investigated whether any differences existed in the yield and molar mass of EPS when milk was fermented at the same acidification rate by 2 strains of S. thermophilus (St-143 and ST-10255y). The type of EPS produced by these 2 strains is different. Milk samples were analyzed for EPS concentration every 30 min during a fermentation period of 270 min (final pH 4.5) by using a modified quantification method, which was faster and validated for its recovery of added EPS. Rheological properties of milks during fermentation were also analyzed using small-strain dynamic oscillatory rheology. For the determination of molar mass, EPS extracts were isolated by ultrafiltration of whey obtained during fermentation of milk to pH values 5.2, 4.9, 4.7, and 4.5, and molar mass was analyzed using size-exclusion chromatography–multi-angle laser light scattering. During fermentation, both strains appeared to start producing significant amounts of EPS after about ～150 min, which corresponded to pH ～5.3, which was close to the point of gelation. During the remainder of the fermentation process (150–270 min), the EPS concentration from strains St-143 and ST-10255y significantly increased from 30 to 72 mg/L and from 26 to 56 mg/L, respectively. The quantity of EPS recovered by our modified method was estimated to represent ～60% of the total EPS added to milk. The molar mass of EPS produced by both strains appeared to slightly decrease during fermentation. At pH 5.2, EPS from St-143 and ST-10255y had molar masses of 2.9 × 10⁶ and 1.4 × 10⁶ g/mol, respectively, which decreased to 1.6 × 10⁶ and 0.8 × 10⁶ g/mol, respectively, when the pH of milk was 4.5. Distinct differences were apparent in the rheological properties of gels fermented by the 2 strains. At the end of fermentation, St-143 fermented milk had weaker gels with storage modulus (G′) value at pH 4.6 of 26 Pa, whereas gels made with ST-10255y were stiffer with a G′ value at pH 4.6 of 82 Pa. For St-143 gels, maximum loss tangent (LT_max) values were higher (0.50) and occurred earlier (at a higher pH value) than the LT_max values (0.46) for gels from ST-10255y strain. Because the fermentation conditions were identical for both strains, the observed changes in rheological properties could be due to the differences in chemical structures and molar mass of the EPS produced by these 2 S. thermophilus strains.     

Effect of fermentation temperature on the properties of exopolysaccharides and the acid gelation behavior for milk fermented by Streptococcus thermophilus strains DGCC7785 and St-143

We investigated the effect of fermenting milk with 2 strains (DGCC7785 and St-143) of Streptococcus thermophilus, which are known to produce different types of exopolysaccharide (EPS) structures. The yields and physical properties of these ropy EPS were monitored during the fermentation of milk at different temperatures. We wanted to understand how these types of EPS properties affected yogurt gelation. Reconstituted skim milk was fermented at 33, 39, or 45°C until pH values reached 5.2, 4.9, 4.7, and 4.5. Molar mass of ropy EPS samples was determined using size exclusion chromatography coupled with multiangle laser light scattering. Rheological properties of fermented milk gels were analyzed using small-strain dynamic oscillatory measurements. In both strains, concentrations of ropy EPS increased during fermentation and at all temperatures. Fermentation times, by both strains, were shortest at 45°C and longest at 33°C. For both strains, molar mass of ropy EPS ranged from 2 to 4 × 10⁶ g/mol during fermentation. A major proteinaceous contaminant that was co-isolated with the ropy EPS fraction by our isolation method was identified as a milk-derived phosphoglycoprotein PP3. Increase in fermentation temperature from 33 to 45°C significantly decreased the storage modulus values (from 170 to 41 Pa) for milk gelled by strain DGCC7785, whereas the gels made with St-143 had very low storage modulus values (11–17 Pa) regardless of fermentation temperatures. For both strains, the values of maximum loss tangent in the milk gels increased with fermentation temperature; the maximum loss tangent occurred at higher pH values when milk was fermented by strain DGCC7785. The specific type of EPS produced appeared to be responsible for the differences in yogurt texture rather than the concentration or molar mass of the EPS.     

Effects of exopolysaccharide-producing strains of Streptococcus thermophilus on technological and rheological properties of set-type yoghurt

This study investigated the effects of two Streptococcus thermophilus strains, ST 285 and ST 1275, on selected technological and rheological characteristics of set-type yoghurt. The strains were selected for their capability to produce distinctly different exopolysaccharides (EPS) and were thus coded as capsular (ST 285) or ropy-capsular (ST 1275). The culture performance and physico-chemical properties of yoghurt were assessed in relation to different fermentation temperatures (30, 37 or 42 °C) and prolonged storage (up to 30 days) at low temperature (4 °C). ST 1275 showed faster growth and acidification rates, resulting in yoghurt with lower syneresis and higher-flow behaviour index, than ST 285. EPS production appeared to be growth associated with the maximum given at growth temperatures of 37 and 42 °C for ST 285 and ST 1275, respectively; however, EPS concentration declined considerably during storage. Prolonged cold storage increased several rheological characteristics of yoghurt including G′, consistency index and hysteresis loop area. A weak correlation between EPS concentration and textural properties of yoghurt was observed.     

Exopolysaccharide-producing Streptococcus thermophilus strains as functional starter cultures in the production of fermented milks

Relationships between exopolysaccharide (EPS) production (amount, molecular mass and sugar composition of the EPS) by different Streptococcus thermophilus strains as a functional starter culture, and textural characteristics (viscosity) of fermented milk and yoghurt have been studied. Five interesting heteropolysaccharide-producing strains have been tested. Both S. thermophilus LY03 and S. thermophilus CH101 produced the highest amounts of EPS and also displayed the highest apparent viscosities in fermented milk. S. thermophilus ST 111 and S. thermophilus STD differed considerably in EPS yields, but not in apparent viscosities of fermented milk. In addition, S. thermophilus ST 111 displayed a high variability in EPS amounts when cultivated in milk. In milk medium, S. thermophilus LY03 produced two heteropolysaccharides, a high-molecular-mass (HMM) EPS and a low-molecular-mass (LMM) EPS of the same composition (Gal/Glu/GalNAc=3.4:1.4:1.0). S. thermophilus ST 111 produced only a HMM-EPS (Gal/Rha=2.5:1.0), while S. thermophilus CH 101 (Gal/Glu=1.0:1.0), S. thermophilus ST 113 (Gal/Glu/Rha/GalNAc=1.7:3.9:1.5:1.0) and S. thermophilus STD (Gal/Glu/Rha/GalNAc=3.5:6.2:1.2:1.0) produced only LMM-EPS. Both HMM-EPS and LMM-EPS solutions (S. thermophilus LY03) demonstrated a pseudoplastic character; HMM-EPS solutions of 0.2% (m/v) displayed a high consistency as well. Although its production of high EPS amounts, S. thermophilus LY03 resulted in relatively thin yoghurts, so that texture values did not directly correlate with EPS production capacity. Once structure/function relationships are known, one can determine the molecular properties of the isolated and purified EPS (molecular size, structural characteristics) from candidate strains to predict their potential in texture formation. For a final selection of interesting EPS-producing starter strains one should test the EPS production under yoghurt manufacturing conditions.     

Fermentation of salmon fillets with a variety of lactic acid bacteria

The influence of five strains of lactic acid bacteria (four Lactobacillus and one Carnobacterium) on the quality of fermented salmon fillets was studied. Best starter growth (increase of more than 1 log in 3 days) and acidification of muscle (e.g. pH reduction of approximately 0.7 units in 5 days) were achieved with the two commercial strains L. sake LAD and L. alimentarius BJ33. pH reduction was consistently lower (e.g. reduction of 0.2 units in 5 days) with C. piscicola 85. Protein breakdown as observed on SDS-PAGE gels was similar for all strains. In contrast, the starter strain did influence texture and colour changes. Fast acidifying strains L. sake LAD and L. alimentarius BJ33 brought about a firmer overall texture and a lighter colour, while softening of flesh occurred in samples processed with C. piscicola 85. Sensory evaluations indicated that samples processed with fast acidifying strains were preferred. L. sake LAD and L. alimentarius BJ33 are regarded as suitable starters for fermentation of salmon fillets.     

Production of exopolysaccharides by Lactobacillus and Bifidobacterium strains of human origin, and metabolic activity of the producing bacteria in milk

This work reports on the physicochemical characterization of 21 exopolysaccharides (EPS) produced by Lactobacillus and Bifidobacterium strains isolated from human intestinal microbiota, as well as the growth and metabolic activity of the EPS-producing strains in milk. The strains belong to the species Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus vaginalis, Bifidobacterium animalis, Bifidobacterium longum, and Bifidobacterium pseudocatenulatum. The molar mass distribution of EPS fractions showed 2 peaks of different sizes, which is a feature shared with some EPS from bacteria of food origin. In general, we detected an association between the EPS size distribution and the EPS-producing species, although because of the low numbers of human bacterial EPS tested, we could not conclusively establish a correlation. The main monosaccharide components of the EPS under study were glucose, galactose, and rhamnose, which are the same as those found in food polymers; however, the rhamnose and glucose ratios was generally higher than the galactose ratio in our human bacterial EPS. All EPS-producing strains were able to grow and acidify milk; most lactobacilli produced lactic acid as the main metabolite. The lactic acid-to-acetic acid ratio in bifidobacteria was 0.7, close to the theoretical ratio, indicating that the EPS-producing strains did not produce an excessive amount of acetic acid, which could adversely affect the sensory properties of fermented milks. With respect to their viscosity-intensifying ability, L. plantarum H2 and L. rhamnosus E41 and E43R were able to increase the viscosity of stirred, fermented milks to a similar extent as the EPS-producing Streptococcus thermophilus strain used as a positive control. Therefore, these human EPS-producing bacteria could be used as adjuncts in mixed cultures for the formulation of functional foods if probiotic characteristics could be demonstrated. This is the first article reporting the physicochemical characteristics of EPS isolated from human intestinal microbiota.     

Nonstarter Lactobacillus strains as adjunct cultures for cheese making: in vitro characterization and performance in two model cheeses

Nonstarter lactic acid bacteria are the main uncontrolled factor in today's industrial cheese making and may be the cause of quality inconsistencies and defects in cheeses. In this context, adjunct cultures of selected lactobacilli from nonstarter lactic acid bacteria origin appear as the best alternative to indirectly control cheese biota. The objective of the present work was to study the technological properties of Lactobacillus strains isolated from cheese by in vitro and in situ assays. Milk acidification kinetics and proteolytic and acidifying activities were assessed, and peptide mapping of trichloroacetic acid 8% soluble fraction of milk cultures was performed by liquid chromatography. In addition, the tolerance to salts (NaCl and KCl) and the phage-resistance were investigated. Four strains were selected for testing as adjunct cultures in cheese making experiments at pilot plant scale. In in vitro assays, most strains acidified milk slowly and showed weak to moderate proteolytic activity. Fast strains decreased milk pH to 4.5 in 8 h, and continued acidification to 3.5 in 12 h or more. This group consisted mostly of Lactobacillus plantarum and Lactobacillus rhamnosus strains. Approximately one-third of the slow strains, which comprised mainly Lactobacillus casei, Lactobacillus fermentum, and Lactobacillus curvatus, were capable to grow when milk was supplemented with glucose and casein hydrolysate. Peptide maps were similar to those of lactic acid bacteria considered to have a moderate proteolytic activity. Most strains showed salt tolerance and resistance to specific phages. The Lactobacillus strains selected as adjunct cultures for cheese making experiments reached 10⁸ cfu/g in soft cheeses at 7 d of ripening, whereas they reached 10⁹ cfu/g in semihard cheeses after 15 d of ripening. In both cheese varieties, the adjunct culture population remained at high counts during all ripening, in some cases overcoming or equaling primary starter. Overall, proximate composition of cheeses with and without added lactobacilli did not differ; however, some of the tested strains continued acidifying during ripening, which was mainly noticed in soft cheeses and affected overall quality of the products. The lactobacilli strains with low acidifying activity showed appropriate technological characteristics for their use as adjunct cultures in soft and semihard cheeses.     

Milk fermentation by functional mixed culture producing nisin Z and exopolysaccharides in a fresh cheese model

A nisin Z-producing strain, Lactococcus lactis subsp. lactis biovar. diacetylactis UL719 and two nisin-sensitive cultures, Lactobacillus rhamnosus RW-9595 M producing exopolysaccharide (EPS), and Lc. lactis subsp. cremoris for acidification, were tested in pure and mixed cultures during milk fermentation. The mixed culture of the three strains showed a higher acidifying capacity at 34°C and 38°C, even though populations of Lc. cremoris were largely reduced compared with pure cultures. Bacteriocin production was 3.1–4.6-fold higher in mixed cultures than for pure cultures of Lc. diacetylactis UL719. These data can be explained by commensalism behavior relying on high proteolytic activity of Lc. cremoris and autolysis and nisin Z-induced lysis. In mixed culture, EPS production was 3-fold lower than for Lb. rhamnosus RW-9595 M pure culture. Our data showed that this strain combination, with nisin-producing and sensitive strains, can be used in mixed cultures for manufacture of fresh cheese with improved functional properties.     

Exopolysaccharide produced by Weissella confusa: Chemical characterisation,rheology and bioactivity

The purpose of this study was to characterise the production of exopolysaccharide (EPS) by indigenous lactic acid bacteria (LAB) isolated from traditional Algerian dairy products and to evaluate their possible use in agri-foods. Among the collection of isolated strains, the strain Weissella confusa (W4) was selected for its ability to produce EPS once exposed to a sucrose culture medium. EPS produced were first isolated with a standardised method and further characterised in terms of molecular size, antioxidant activity, and rheological properties. Its direct implication in the texture and syneresis of acid milk gel was evaluated offering interesting industrial applications for its use during processes dealing with dairy products.     

Application of ruthenium red and colloidal gold-labeled lectin for the visualization of bacterial exopolysaccharides in Cheddar cheese matrix using transmission electron microscopy

The objective of the present study was to develop a methodology for direct observation of capsular and ropy strains and their exopolysaccharides (EPS) in a Cheddar cheese matrix. Cheddar cheeses with 50% reduced fat were made from milk containing 1.7% fat using mixed starter culture containing either capsule-forming Lactococcus lactis subsp. cremoris (SMQ-461) or ropy L. lactis subsp. cremoris (JRF-1) strains. Control cheese was made using the EPS-negative L. lactis subsp. cremoris (RBL132) strain. Following cheese pressing, samples were taken from each cheese treatment and examined by transmission electron microscopy (TEM). Samples were divided into two series: the first was prepared following the conventional methods (involving fixation, post fixation, dehydration and embedding in resin) and the second with added ruthenium red at 0.15% (w/v) during the fixation, post fixation and washing procedures. Gold-labeled lectin was also used for the visualization and localization of EPS in cheese matrix. Electron micrographs showed that ruthenium red makes it possible to visualize and enhance the resolution of the EPS in a Cheddar matrix compared with the conventional method. The EPS layer of the capsular strain appeared regular and evenly distributed around the cell, whereas the cell-associated EPS layer produced by the ropy strain was longer, more irregular (having a filamentous structure) and unevenly surrounded the cell. EPS released from the ropy strain appeared to form a network-like structure located principally in whey pockets and appeared to interact with the casein matrix and fat globule membrane. Labeling EPS by lectin conjugated to colloidal gold could only be performed with conventional preparation of cheese samples and appeared to react only with the cell surface rather than with liberated EPS. Besides their ability to bind water and increase cheese yield, capsular and ropy strains used in this study appear to have potential autolytic characteristics, which may have an impact on cheese proteolysis, texture and flavor quality.     

Short communication: impact of pH and temperature on the acidifying activity of Carnobacterium maltaromaticum

The acidifying activity of Carnobacterium maltaromaticum LMA28, a strain isolated from French soft cheese, was studied in trypticase soy broth with yeast extract (TSB-YE) medium and in milk. In TSB-YE supplemented with lactose, glucose, or galactose, lactose and glucose were metabolized with a maximum growth rate of 0.32 h⁻¹ and galactose was not metabolized. During hydrolysis of lactose, the galactose moiety was not excreted. The major product was l(+) lactic acid, with no significant difference in the lactic acid yield. Glucose was not completely metabolized because cell growth stopped when pH values reached an average of 5.0. In sterilized UHT milk, the addition of 1 g/L of YE enhanced its coagulation. Compared with commercial starter lactic acid bacteria such as Lactococcus lactis DSMZ 20481 or Streptococcus thermophilus INRA 302, Carnobacterium maltaromaticum LMA 28 was shown to be a slow acidifying strain. However, in spite of this weak acidifying ability, C. maltaromaticum LMA 28 can sustain low pH values in coculture with Lc. lactis DSMZ 20481 or S. thermophilus INRA 302. The individual and interactive effects of initial pH values (5.2 to 8.0) and incubation temperatures (23 to 37°C) on acidifying activity were studied by response surface methodology. The 3 strains displayed different behaviors depending on pH and temperature. The psychrotrophic lactic acid strain C. maltaromaticum LMA 28 was able to grow at alkaline pH values and during storage conditions. It could be used as a potential ripening flora in soft cheese.     

Performance of <Emphasis Type="Italic">Lactobacillus sanfranciscensis</Emphasis> TMW 1.392 and its levansucrase deletion mutant in wheat dough and comparison of their impact on bread quality

The performance of Lactobacillus sanfranciscensis TMW 1.392 and its levansucrase deletion mutant TMW 1.392 Δlev in wheat dough was compared. The effects of both strains on dough and bread characteristics were determined in order to find benchmarks for in situ production of exopolysaccharides (EPS). Growth and acidification were lower in doughs prepared with the Δlev mutant than in those employing the wild type. Extensogram resistance of the dough was reduced and extensibility increased with the addition of L. sanfranciscensis levan. Added EPS positively influenced water absorption, bread volume and firming of the crumb. In situ production of EPS was not sufficient to achieve the same positive effects of EPS, as they partially overlapped with effects resulting from enhanced acidification. Control doughs were made to separate effects of predough, EPS and different metabolism/acidification. High acetic acid levels decreased extensibility and volume. High lactic acid levels negatively influenced crumb hardness and firming kinetics. The use of knock out mutants proved helpful to judge overall performance of a strain, although the interpretation of specific effects must consider all changes in its metabolism.     

Effect of exopolysaccharides and proteolytic activity of Lactococcus lactis subsp. cremoris strains on the viscosity and structure of fermented milks

Two exopolysaccharide (EPS)-producing Lactococcus lactis subsp. cremoris strains (B35 and B891) were used to study the effect of the kinetics of EPS production and bacterial proteolytic activity on the structure of milk gels and the viscosity of stirred milk gels. Strains were grown at 20 °C in milk containing either yeast extract or casitone and at 30 °C in either milk alone or milk containing casitone. Lactococcal counts, pH decrease and production and molecular characteristics (molar mass and radius of gyration) of both EPSs were followed during milk fermentation. The level of proteolysis in the fermented milks was determined after 24 h of incubation. The results obtained showed that the yield of EPS and the timing of EPS production during milk-gel formation were the most important factors that influenced the structure of the milk gels and the viscosity of the stirred product. The proteolytic activity of the strains did not seem to play any significant role.     

Use of microparticulated whey protein concentrate,exopolysaccharide-producing Streptococcus thermophilus, and adjunct cultures for making low-fat Italian Caciotta-type cheese

Low-fat Caciotta-type cheeses were manufactured with partially skim milk (fat content of ~0.3%) alone (LFC); with the supplementation of 0.5% (wt/vol) microparticulated whey protein concentrate (MWPC) (LFC-MWPC); with MWPC and exopolysaccharides (EPS)-producing Streptococcus thermophilus ST446 (LFC-MWPC-EPS); and with MWPC, EPS-producing strain ST446, and Lactobacillus plantarum LP and Lactobacillus rhamnosus LRA as adjunct cultures (LFC-MWPC-EPS-A). The non-EPS-producing isogenic variant Streptococcus thermophilus ST042 was used for making full-fat Caciotta-type cheese (FFC), LFC, and LFC-MWPC. Cheeses were characterized based on compositional, microbiological, biochemical, texture, volatile components (purge and trap, and solid-phase microextraction coupled with gas chromatography-mass spectrometry), and sensory analyses. Compared with FFC and LFC (51.6 ± 0.7 to 53.0 ± 0.9%), the other cheese variants retained higher levels of moisture (60.5 ± 1.1 to 67.5 ± 0.5%). The MWPC mainly contributed to moisture retention. Overall, all LFC had approximately one-fourth (22.6 ± 0.8%) of the fat of FFC. Hardness of cheeses slightly varied over 7 d of ripening. Microbial EPS positively affected cheese texture, and the texture of LFC without MWPC or microbial EPS was excessively firm. Free amino acids were at the highest levels in LFC treatments (2,705.8 ± 122 to 3,070.4 ± 123 mg/kg) due to the addition of MWPC and the peptidase activity of adjunct cultures. Aldehydes, alcohols, ketones, sulfur compounds, and short- to medium-chain carboxylic acids differentiated LFC variants and FFC. The sensory attributes pleasant to taste, intensity of flavor, overall acceptability, and pleasant to chew variously described LFC-MWPC-EPS and LFC-MWPC-EPS-A. Based on the technology options used, low-fat Caciotta-type cheese (especially ripened for 14 d) has promising features to be further exploited as a suitable alternative to the full-fat variant.     

Enzymatic activity of lactic acid bacteria (with antimicrobial properties) isolated from a traditional Spanish cheese

Twenty-four strains of lactic acid bacteria (LAB) isolated from a traditional Spanish cheese (Genestoso cheese) were evaluated for their enzymatic activities (acidifying and proteolytic abilities and carboxypeptidase, aminopeptidase, dipeptidase, caseinolytic and esterase activities), in order to select indigenous strains of technical interest for the manufacture of cheese. These strains were selected on the basis of their antimicrobial activity relative to five reference strains and were identified as Lactococcus lactis subsp. lactis (thirteen strains), Leuconostoc mesenteroides (two strains), Leuconostoc pseudomesenteroides (one strain), Lactobacillus paracasei (two strains), Lactobacillus plantarum (one strain) and Enterococcus faecalis (five strains).     

Gelation and resistance to shearing of fermented milk: Role of exopolysaccharides

Milk was fermented with the exopolysaccharide-producing (EPS⁺) strains Lactococcus lactis subsp. cremoris, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and with the non-EPS-producing strain (EPS⁻) L. lactis subsp. cremoris. The kinetics of gelation and the behaviour of set fermented milk during and after shearing were studied using rheometry and confocal scanning laser microscopy. The time of gelation of milk depended on the kinetics of acidification of strains whereas the pH of gelation depended mostly on the presence of exopolysaccharides (EPS). In set fermented milk with EPS⁺ strains, bacteria were observed in protein-free areas likely filled with EPS. Phase-separated EPS and caseins contributed to induce the gelation of fermented milk at pH 5.6. The high resistance to shearing of milk fermented with the EPS⁺ strain L. lactis subsp. cremoris might be due to the negative charge of the exopolysaccharide allowing an attractive interaction with caseins.     

Selection and properties of Streptococcus thermophilus mutants deficient in urease

Natural variations of the urea content of milk have a detrimental effect on the regularity of acidification by Streptococcus thermophilus strains used in dairy processes. The aim of the present study was to select urease-deficient mutants of S. thermophilus and to investigate their properties. Using an improved screening medium on agar plates, mutants were selected from 4 different parent strains after mutagen treatment and by spontaneous mutation. Most mutants were stable and had a phage sensitivity profile similar to that of their parent strain. Some of them contained detrimental secondary mutations, as their acidifying activity was lower than that of the parent strain cultivated in the presence of the urease inhibitor flurofamide. The proportion of this type of mutant was much lower among spontaneous mutants than among mutants selected after mutagen treatment. Utilization of urease-deficient mutants in dairy processes may have several advantages, such as an increase in acidification, an improved regularity of acidification, and a lower production of ammonia in whey.