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1.
J Food Sci ; 89(5): 2546-2556, 2024 May.
Article in English | MEDLINE | ID: mdl-38578148

ABSTRACT

2'-Fucosyllactose (2'-FL) is postulated to provide health benefits and promote the growth of probiotics. This work was undertaken to study the effects of 2'-FL on the viability of starter cultures and Bifidobacterium strains of human origin in yogurt during refrigerated storage. Yogurts were produced containing 2'-FL (0 or 2 g/L) and Bifidobacterium strains of human origin (Bifidobacterium longum subsp. longum BB536 or Bifidobacterium longum subsp. infantis ATCC 15697) at a concentration of at least 109 CFU/mL. All yogurts were stored at 4°C for 5 weeks. Results showed that 2'-FL was stable in yogurts for at least 5 weeks of cold storage, and the addition of 2'-FL did not significantly alter yogurt fermentation parameters, associated metabolites, and the viability of mixed yogurt starter cultures and Bifidobacterium strains (p > 0.05). The addition of bifidobacteria had a negative impact (p < 0.05) on the survival rate of starter cultures, Streptococcus thermophilus and Lactobacillus delbureckii subsp. bulgaricus. Meanwhile, it is difficult to maintain a high survival rate of bifidobacteria in final yogurt products, and the addition of 2'-FL could not enhance the viability of bifidobacteria. B. longum BB536 survived at a level higher than 106 CFU/g for 28 days, while B. infantis ATCC15697 maintained this level for only 7 days. In summary, this study has shown the impact of 2'-FL and bifidobacterial species on yogurt properties, and results suggest that it is promising to use 2'-FL in yogurt products as a prebiotic. PRACTICAL APPLICATION: Yogurt is known for its beneficial effects on human health and nutrition. This study reported the production of symbiotic yogurt containing bifidobacteria and 2'-fucosyllactose (2'-FL) as a functional food for specified health uses. The viability of yogurt starter cultures and probiotic bifidobacterial strains was analyzed in this study. Moreover, this research demonstrated that 2'-FL could be added to yogurt without affecting the characteristics of yogurt significantly.


Subject(s)
Bifidobacterium , Fermentation , Food Storage , Probiotics , Trisaccharides , Yogurt , Yogurt/microbiology , Trisaccharides/pharmacology , Bifidobacterium/growth & development , Humans , Food Storage/methods , Refrigeration , Streptococcus thermophilus/growth & development , Microbial Viability , Food Microbiology , Colony Count, Microbial
2.
Food Microbiol ; 95: 103672, 2021 May.
Article in English | MEDLINE | ID: mdl-33397607

ABSTRACT

Enumeration and isolation of Streptococcus salivarius subsp. thermophilus from cheese is challenging, due to the relatively high number of species it may host. We describe medium SPY9.3 for the cultivation of S. salivarius subsp. thermophilus from cheese. The medium and related incubation conditions (SPY) was compared with 2 other protocols, M17 and ST: sensitivity was assessed by parallel cultivation of 55 strains of S. salivarius subsp. thermophilus, and selectivity by (i) parallel cultivation of 60 strains belonging to 20 different non-target species and sub-species and (ii) isolating bacteria from 3 raw-milk cheeses. Colony counts were similar on SPY9.3 and M17 (mean difference 0.07 log(cfu/mL), p > 0.001) and significantly higher on ST than on M17 and SPY9.3 (mean differences 0.42 and 0.48 log(cfu/mL), respectively, p < 0.001). SPY was more specific than ST and M17, with respectively 20%, 40%, and 50% of the investigated non-target species able to grow. S. salivarius subsp. thermophilus, Enterococcus spp., and Staphylococcus aureus were indistinguishable using all 3 protocols. Only SPY avoided growth of Lactobacillus delbrueckii subsp. lactis. Finally, ST and SPY displayed higher recoveries of S. salivarius subsp. thermophilus colonies from cheese than M17 (5.6, 5.5, and 3.0 adjusted log(cfu/mL), respectively) and the lowest proportion of non-specific isolates. The protocol described here and based on SPY9.3 presents a promising alternative to existing protocols for the enumeration and isolation of S salivarius subsp. thermophilus from cheese or other complex fermented products.


Subject(s)
Cheese/microbiology , Culture Media/metabolism , Food Microbiology/methods , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , Animals , Cattle , Colony Count, Microbial , Culture Media/chemistry , Food Microbiology/instrumentation , Milk/microbiology , Streptococcus thermophilus/genetics
3.
J Dairy Sci ; 104(2): 1484-1493, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33309375

ABSTRACT

Camel milk, similar to cow milk, contains all of the essential nutrients as well as potentially health-beneficial compounds with anticarcinogenic, antihypertensive, and antioxidant properties. Camel milk has been used for the treatment of allergies to cow milk, diabetes, and autism. Camel milk helps decrease cholesterol levels in blood and improves metabolism. One of the most desirable food tastes is sweetness. However, the excessive ingestion of sugar negatively affects human health. Monk fruit sweetener is a natural, 0-calorie sweetener with many health-beneficial functions. Monk fruit sweetener helps decrease symptoms of asthma and diabetes, prevents oxidation and cancer, protects the liver, regulates immune function, and lowers glucose levels. Monk fruit sweetener is 100 to 250 times sweeter than sucrose. The objective of this study was to examine the influence of different concentrations of monk fruit sweetener on the physicochemical properties and microbiological counts of drinking yogurt made from camel milk. Camel milk drinking yogurt was produced with 0, 0.42, 1.27, and 2.54 g/L of monk fruit sweetener and stored for 42 d. The physicochemical characteristics and microbiological counts of yogurts were measured at d 1, 7, 14, 21, 28, 35, and 42. For the physicochemical characteristics, pH, titratable acidity, viscosity, and color [lightness-darkness (L*), red-green axis (a*), yellow-blue axis (b*), chroma (C*), and hue angle (h*)] values were evaluated. The counts of Streptococcus thermophilus, Lactobacillus bulgaricus, Lactobacillus acidophilus, coliforms, and yeast and mold were determined. Three replications were conducted. The sweetener addition significantly influenced pH, viscosity, and color (a*, b*, C*, and h*) values. Control samples had significantly higher pH values, lower viscosity, lower b* and C* values, and higher h* values than the samples with 1.27 and 2.54 g/L of monk fruit sweetener. Growth of S. thermophilus, L. bulgaricus, and probiotic culture L. acidophilus was not affected by the incorporation of monk fruit sweetener. Monk fruit sweetener can be added in camel milk yogurts as a health-beneficial 0-calorie sweetener.


Subject(s)
Lactobacillus acidophilus/growth & development , Lactobacillus delbrueckii/growth & development , Milk/chemistry , Probiotics , Streptococcus thermophilus/growth & development , Sweetening Agents/chemistry , Yogurt/microbiology , Animals , Antioxidants/analysis , Camelus , Chemical Phenomena , Female , Fermentation , Fruit/chemistry , Yogurt/analysis
4.
Food Chem ; 341(Pt 1): 128163, 2021 Mar 30.
Article in English | MEDLINE | ID: mdl-33035853

ABSTRACT

Lactic acid bacteria fermentation is a safe and green technology that can modify the function of food ingredients (including proteins). In this article, egg yolks were subjected to fermentation with commercial lactic acid bacteria for 0, 3, 6 and 9 h, respectively. After fermentation treatment, the microbial composition has changed obviously (Streptococcus thermophilus increased significantly). The free sulfhydryl group (SH) contents and surface hydrophobicity of egg yolk proteins were significantly reduced. The rheological results indicated that the treated egg yolks possessed a decreased apparent viscosity. Correspondingly, the emulsifying activity of egg yolk was enhanced from 9.07 to 19.55, 23.40 and 24.61 m2/g for 3, 6 and 9 h of fermentation, respectively. And the emulsifying stability reached the maximum after 3 h of fermentation. This study investigated the relationship between structure and properties of yolk proteins, and showed that lactic acid fermentation endued egg yolk with better emulsifying properties.


Subject(s)
Egg Proteins/chemistry , Egg Yolk/chemistry , Egg Yolk/microbiology , Lactobacillales/metabolism , Animals , Egg Proteins/analysis , Egg Proteins/metabolism , Emulsions/chemistry , Fermentation , Food Microbiology , Hydrophobic and Hydrophilic Interactions , Rheology , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , Sulfhydryl Compounds/chemistry , Viscosity
5.
Food Chem ; 343: 128553, 2021 May 01.
Article in English | MEDLINE | ID: mdl-33176956

ABSTRACT

This study investigated the bioconversion and bioaccessibility of soy isoflavones produced in sogurt fermented with S. thermophilus and L. bulgaricus during in vitro digestion. The highest survivability of S. thermophilus (6.49 log cfu/mL) and L. bulgaricus (6.48 log cfu/mL) was in oral phase. In gastric phase, the total aglycones of sogurt (26.73 g/L) increased up to 20 times than control (1.21 g/L), with a significant increase in daidzein (17.05 g/L) and genistein (9.68 g/L). Addition of 8U of ß-glucosidase into soymilk significantly increased the conversion of isoflavone in ENTII (daidzein: 0.46 g/L; genistein: 0.18 g/L) than in ENTI (daidzein: 0.33 g/L; genistein: 0.20 g/L). The particle size analysis and confocal micrographs of digesta also suggest the size of fat and protein in gastric phase to be smaller than in intestinal phase. The results indicate the prospective to develop soy-based fermented products capable of releasing high isoflavone in the digestive system.


Subject(s)
Isoflavones/metabolism , Yogurt/analysis , Chromatography, High Pressure Liquid , Chromatography, Reverse-Phase , Digestion , Genistein/metabolism , Isoflavones/analysis , Lactobacillus delbrueckii/growth & development , Lactobacillus delbrueckii/metabolism , Soy Milk/metabolism , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , beta-Glucosidase/metabolism
6.
Int J Food Microbiol ; 335: 108903, 2020 Dec 16.
Article in English | MEDLINE | ID: mdl-33065381

ABSTRACT

Societal demand for plant-based foods is increasing. In this context, soya products fermented using lactic acid bacteria (LAB) are appealing because of their potential health and nutritional benefits. The thermophilic LAB Streptococcus thermophilus is an essential starter species in the dairy industry. However, while its physiology is well characterized, little is known about its general metabolic activity or its techno-functional properties when it is grown in soya milk. In this study, S. thermophilus LMD-9 growth, sugar production, and lactic acid production in soya milk versus cow's milk were measured. Additionally, the main metabolic pathways used by the bacterium when growing in soya milk were characterized using a proteomic approach. Streptococcus thermophilus LMD-9 growth decreased soya milk pH, from 7.5 to 4.9, in 5 h. During fermentation, acidification thus occurred in tandem with lactate production and increasing population size (final population: 1.0 × 109 CFU/ml). As growth proceeded, sucrose was consumed, and fructose was produced. The proteomic analysis (LC-MS/MS) of the strain's cytosolic and cell envelope-associated proteins revealed that proteins related to amino acid transport and nitrogen metabolism were the most common among the 328 proteins identified (63/328 = 19.2% of total proteins). The cell-wall protease PrtS was present, and an LMD-9 deletion mutant was constructed by interrupting the prtS gene (STER_RS04165 locus). Acidification levels, growth levels, and final population size were lower in the soya milk cultures when the ΔprtS strain versus the wild-type (wt) strain was used. The SDS-PAGE profile of the soluble proteins in the supernatant indicated that soya milk proteins were less hydrolyzed by the ΔprtS strain than by the wt strain. It was discovered that S. thermophilus can grow in soya milk by consuming sucrose, can hydrolyze soya proteins, and can produce acidification levels comparable to those in cow's milk. This study comprehensively examined the proteomics of S. thermophilus grown in soya milk and demonstrated that the cell-wall protease PrtS is involved in the LAB's growth in soya milk and in the proteolysis of soya proteins, which are two novel findings. These results clarify how S. thermophilus adapts to soya milk and can help inform efforts to develop new fermented plant-based foods with better-characterized biochemical and microbiological traits.


Subject(s)
Bacterial Proteins/metabolism , Serine Endopeptidases/metabolism , Soy Milk/metabolism , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , Animals , Fermentation , Lactic Acid/analysis , Lactic Acid/metabolism , Metabolic Networks and Pathways , Milk/chemistry , Milk/metabolism , Milk/microbiology , Milk Proteins/metabolism , Nitrogen/metabolism , Proteomics , Soy Milk/chemistry , Soybean Proteins/metabolism , Streptococcus thermophilus/enzymology , Sucrose/metabolism , Sugars/analysis , Sugars/metabolism
7.
Genes (Basel) ; 11(9)2020 09 19.
Article in English | MEDLINE | ID: mdl-32961685

ABSTRACT

In streptococci, intracellular quorum sensing pathways are based on quorum-sensing systems that are responsible for peptide secretion, maturation, and reimport. These peptides then interact with Rgg or ComR transcriptional regulators in the Rap, Rgg, NprR, PlcR, and PrgX (RRNPP) family, whose members are found in Gram-positive bacteria. Short hydrophobic peptides (SHP) interact with Rgg whereas ComS peptides interact with ComR regulators. To date, in Streptococcus thermophilus, peptide secretion, maturation, and extracellular fate have received little attention, even though this species has several (at least five) genes encoding Rgg regulators and one encoding a ComR regulator. We studied pheromone export in this species, focusing our attention on PptAB, which is an exporter of signaling peptides previously identified in Enterococcus faecalis, pathogenic streptococci and Staphylococcus aureus. In the S. thermophilus strain LMD-9, we showed that PptAB controlled three regulation systems, two SHP/Rgg systems (SHP/Rgg1358 and SHP/Rgg1299), and the ComS/ComR system, while using transcriptional fusions and that PptAB helped to produce and export at least three different mature SHPs (SHP1358, SHP1299, and SHP279) peptides while using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using a deep sequencing approach (RNAseq), we showed that the exporter PptAB, the membrane protease Eep, and the oligopeptide importer Ami controlled the transcription of the genes that were located downstream from the five non-truncated rgg genes as well as few distal genes. This led us to propose that the five non-truncated shp/rgg loci were functional. Only three shp genes were expressed in our experimental condition. Thus, this transcriptome analysis also highlighted the complex interconnected network that exists between SHP/Rgg systems, where a few homologous signaling peptides likely interact with different regulators.


Subject(s)
ATP-Binding Cassette Transporters/metabolism , Bacterial Proteins/metabolism , Peptide Fragments/metabolism , Proteome/analysis , Quorum Sensing , Streptococcus thermophilus/metabolism , Transcriptome , ATP-Binding Cassette Transporters/genetics , Bacterial Proteins/genetics , Chromatography, Liquid , Gene Expression Regulation, Bacterial , Streptococcus thermophilus/genetics , Streptococcus thermophilus/growth & development , Tandem Mass Spectrometry
8.
J Food Sci ; 85(10): 3450-3458, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32901954

ABSTRACT

A plain symbiotic almond yogurt-like product was formulated and developed using a plant-based starter YF-L02 (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus supplemented with Lactobacillus acidophilus, Lactobacillus paracasei, and Bifidobacterium animalis) and inulin; 0.6% polymerized whey protein (PWP), 0.3% pectin, and 0.05% xanthan gum were optimized for the formula of the almond yogurt alternative. Two groups with/without calcium citrate and vitamin D2 were prepared and analyzed for chemical composition, changes in pH, viscosity, and probiotic survivability during storage at 4 °C for 10 weeks. The results showed that (1) over 10 weeks storage, the differences in the pH, viscosity, and probiotic survivability between the control and the fortified samples were not significant (P > 0.05); (2) the pH of both yogurt samples decreased 0.2 units while their viscosity slightly increased during storage; (3) the populations of L. paracasei and B. animalis remained above 106 cfu/g during the storage, whereas the population of L. acidophilus decreased dramatically during the first 4 weeks, especially the control group; (4) the microstructure was examined by scanning electron microscopy, revealing a compact and denser gel structure formed by 0.6% PWP with the presence of 0.3% pectin and 0.05% xanthan gum. In conclusion, PWP might be a proper gelation agent for the formulation of symbiotic almond yogurt alternative. PRACTICAL APPLICATION: In this study, polymerized whey protein was used as a gelation agent to formulate symbiotic almond yogurt alternatives with comparable physical texture and probiotic survivability to dairy yogurt during storage. This technology may be used for the development of plant-based fermented foods.


Subject(s)
Lactobacillus acidophilus/growth & development , Lactobacillus delbrueckii/growth & development , Probiotics/chemistry , Prunus dulcis/chemistry , Streptococcus thermophilus/growth & development , Whey Proteins/chemistry , Yogurt/analysis , Fermentation , Gels/chemistry , Gels/metabolism , Inulin/chemistry , Inulin/metabolism , Lactobacillus acidophilus/metabolism , Lactobacillus delbrueckii/metabolism , Microbial Viability , Pectins/chemistry , Pectins/metabolism , Polymerization , Prunus dulcis/metabolism , Prunus dulcis/microbiology , Streptococcus thermophilus/metabolism , Viscosity , Whey Proteins/metabolism , Yogurt/microbiology
9.
Appl Microbiol Biotechnol ; 104(14): 6315-6323, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32462242

ABSTRACT

ß-galactosidases, commonly referred to as lactases, are used for producing lactose-free dairy products. Lactases are usually purified from microbial sources, which is a costly process. Here, we explored the potential that lies in using whole cells of a food-grade dairy lactic acid bacterium, Streptococcus thermophilus, as a substitute for purified lactase. We found that S. thermophilus cells, when treated with the antimicrobial peptide nisin, were able to hydrolyze lactose efficiently. The rate of hydrolysis increased with temperature; however, above 50 °C, stability was compromised. Different S. thermophilus strains were tested, and the best candidate was able to hydrolyze 80% of the lactose in a 50 g/L solution in 4 h at 50 °C, using only 0.1 g/L cells (dry weight basis). We demonstrated that it was possible to grow the cell catalyst on dairy waste, and furthermore, that a cell-free supernatant of a culture of a nisin-producing Lactococcus lactis strain could be used instead of purified nisin, which reduced cost of use significantly. Finally, we tested the cell catalysts in milk, where lactose also was efficiently hydrolyzed. The method presented is natural and low-cost, and allows for production of clean-label and lactose-free dairy products without using commercial enzymes from recombinant microorganisms. KEY POINTS: • Nisin-permeabilized Streptococcus thermophilus cells can hydrolyze lactose efficiently. • A low-cost and more sustainable alternative to purified lactase enzymes. • Reduction of overall sugar content. • Clean-label production of lactose-free dairy products.


Subject(s)
Lactase/metabolism , Lactobacillales/metabolism , Animals , Cell Membrane Permeability/drug effects , Culture Media , Hydrolysis , Lactobacillales/growth & development , Lactococcus lactis/growth & development , Lactococcus lactis/metabolism , Lactose/analysis , Lactose/metabolism , Milk/chemistry , Milk/microbiology , Nisin/metabolism , Nisin/pharmacology , Streptococcus thermophilus/drug effects , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , Temperature
10.
J Sci Food Agric ; 100(10): 3902-3909, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32323334

ABSTRACT

BACKGROUND: This study aimed to evaluate the protective effect of different biopolymer systems on the viability of two probiotics (Lactobacillus rhamnosus and Streptococcus thermophilus) during storage and in vitro digestion. Methylcellulose (MC), sodium alginate (SA), and whey protein (WP)-based structures were designed and characterized in terms of pH, rheological properties, and visual appearance. RESULTS: The results highlighted that the WP-system ensured probiotic protection during both storage and in vitro digestion. This result was attributed to a combined effect of the physical barrier offered by the protein gel network and whey proteins as a nutrient for microbes. On the other hand, surprisingly, the viscous methylcellulose-based system was able to guarantee good microbial viability during storage. However, this was not confirmed during in vitro digestion. The opposite results were obtained for sodium alginate beads. CONCLUSION: The results suggest that the capacity of a polymeric structure to protect probiotic bacteria is a combination of structural organization and system formulation. © 2020 Society of Chemical Industry.


Subject(s)
Biopolymers/chemistry , Lacticaseibacillus rhamnosus/growth & development , Probiotics/chemistry , Streptococcus thermophilus/growth & development , Alginates/chemistry , Digestion , Lacticaseibacillus rhamnosus/chemistry , Microbial Viability , Models, Biological , Streptococcus thermophilus/chemistry , Whey Proteins
11.
J Dairy Sci ; 103(6): 5030-5042, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32307174

ABSTRACT

In this study, we modified reinforced clostridial medium (RCM) to selectively enumerate and isolate Lactobacillus delbrueckii ssp. bulgaricus, a probiotic and important starter culture in the dairy industry. The disparity in the reported carbohydrate fermentation pattern of L. delbrueckii ssp. bulgaricus was used to develop a growth medium not only selective for L. delbrueckii ssp. bulgaricus but significantly inhibitory to the growth of other lactic acid bacteria. A recently modified RCM (mRCM) was optimized for this study by the addition of 0.5% fructose, 0.5% dextrose, 1% maltose, and 0.25% sodium pyruvate while replacing lactose as a carbohydrate source. The cell recovery and bacterial counts of L. delbrueckii ssp. bulgaricus in tested products (pure L. delbrueckii ssp. bulgaricus strains, starter culture, probiotic supplements, and yogurt) using our mRCM with sodium pyruvate (mRCM-PYR) were significantly higher than in the recently modified RCM and the common de Man, Rogosa, and Sharpe (MRS) culture medium. The growth of other lactic acid bacteria (Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus reuteri) and Bifidobacteria was retarded in this modified medium compared with their growth in MRS and mRCM. This result is a significant improvement in the enumeration and differentiation of L. delbrueckii ssp. bulgaricus in mRCM-PYR compared with the results in MRS and mRCM where the high background growth of similar species interferes with the accuracy of bacterial population counts. Our results thus suggest that mRCM-PYR could be recommended as a reliable alternative growth medium for the selective enumeration and isolation of L. delbrueckii ssp. bulgaricus in a mixed culture.


Subject(s)
Clostridium , Culture Media , Lactobacillus delbrueckii/isolation & purification , Animals , Bifidobacterium/growth & development , Fermentation , Lactobacillales/growth & development , Lactobacillus acidophilus/growth & development , Lactobacillus delbrueckii/growth & development , Limosilactobacillus reuteri/growth & development , Lacticaseibacillus rhamnosus/growth & development , Probiotics , Streptococcus thermophilus/growth & development , Yogurt
12.
Carbohydr Polym ; 236: 116019, 2020 May 15.
Article in English | MEDLINE | ID: mdl-32172839

ABSTRACT

In the dairy industry, exopolysaccharides (EPS) produced in situ from lactic acid bacteria are of great interest because of their contribution to product texture. Some EPS cause ropiness which might be linked to specific physical and chemical EPS properties. EPS show a broad variety of chemical structures and, because analysis is rather complex, it is still a major challenge to establish structure-function relationships. The aim of this study was to produce EPS with different degree of ropiness, perform in-depth structural elucidations and relate this information to their behaviour in aqueous solutions. After cultivation of Streptococcus thermophilus DGCC7919 and Lactococcus lactis LL-2A and subsequent EPS isolation, both EPS showed similar macromolecular properties, but pronounced differences in monosaccharide composition and glycosidic linkages. Our data suggests that mainly the side chains in the EPS from LL-2A might be responsible for a higher ropiness than that observed for EPS from DGCC7919.


Subject(s)
Polysaccharides, Bacterial/chemistry , Bioreactors , Carbohydrate Sequence , Fermentation , Glucose/metabolism , Lactococcus lactis/chemistry , Lactococcus lactis/growth & development , Lactococcus lactis/metabolism , Lactose/metabolism , Polysaccharides, Bacterial/biosynthesis , Streptococcus thermophilus/chemistry , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism
13.
Sci Rep ; 10(1): 807, 2020 01 21.
Article in English | MEDLINE | ID: mdl-31964939

ABSTRACT

Streptococcus thermophilus ASCC 1275 has two chain length determining genes - epsC and epsD- in its eps gene cluster, and produces two times more EPS in sucrose medium than that in glucose and lactose. Hence, we investigated the influence of sugars (glucose, sucrose and lactose), at log phase (5 h) and stationary phase (10 h), on the global proteomics of S. thermophilus 1275 to understand the differentially expressed proteins (DEPs) during EPS production using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. Among 98 DEPs in sucrose medium, most of them were mapped into EPS biosynthesis pathway and other related metabolisms. There was an upregulation of several proteins involved in sugar transport (phosphoenolpyruvate (PEP) phosphotransferase system), EPS assembly (epsG1D) and amino acid metabolism (methionine, cysteine/arginine metabolism) in sucrose medium. This study showed that increased EPS production in S. thermophilus 1275 requires a well-co-ordinated regulation of pathway involved in both EPS assembly and amino acid metabolism along with the availability of sugars. Thus, it provided valuable insights into the biosynthesis and regulation of EPS in S. thermophilus 1275, and potential gene targets for understanding high-EPS strains.


Subject(s)
Bacterial Proteins/metabolism , Polysaccharides, Bacterial/biosynthesis , Streptococcus thermophilus/metabolism , Amino Acids/metabolism , Bacterial Proteins/genetics , Biological Transport , Polysaccharides, Bacterial/genetics , Proteomics/methods , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Streptococcus thermophilus/growth & development , Sugars/metabolism
14.
Food Microbiol ; 87: 103383, 2020 May.
Article in English | MEDLINE | ID: mdl-31948624

ABSTRACT

Thermophilic and mesophilic lactic acid bacteria (LAB), such as Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, and Lactococcus lactis, play a crucial role in the technological and sensory quality of Mozzarella cheese. In this study, the safety (genes encoding virulence factors and antibiotic resistance) and acidifying activity of autochthonous S. thermophilus cultures were evaluated in order to choose the most suitable strain for industrial application. The safe and good acidifying culture was tested in two buffalo Mozzarella cheese batches: Mozzarella cheeses produced with autochthonous culture (SJRP107) and commercial culture (STM5). The cultivable LAB was evaluated by culture-dependent method (plate counting) and the quantification of S. thermophilus cultures (commercial and autochthonous) were evaluated by culture-independent method RealT-qPCR (real-time quantitative polymerase chain reaction). The texture, physicochemical and proteolytic properties of the Mozzarella cheeses were similar for both batches. The nonstarter LAB count was higher during manufacture than in the storage, and the RealT-qPCR indicated the presence of S. thermophilus culture until the end of storage. S. thermophilus SJRP107 presented high potential for safety application in the production of Mozzarella cheese. Furthermore, considering the culture characteristics and their relationship with product quality, further studies could be helpful to determine their effect on the sensory characteristics of the cheese.


Subject(s)
Cheese/microbiology , Milk/microbiology , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , Animals , Buffaloes , Cheese/analysis , Consumer Product Safety , Fermentation , Food Microbiology , Humans , Streptococcus thermophilus/genetics , Streptococcus thermophilus/isolation & purification , Taste
15.
Int J Food Microbiol ; 307: 108277, 2019 Oct 16.
Article in English | MEDLINE | ID: mdl-31404779

ABSTRACT

This work was carried out to pursue a double objective: to improve the hygienic safety of cheeses produced from raw ewes' milk; and to produce a new typology of raw ewes' milk through the application of "Grana" technology for which the name "Gran Ovino" was chosen. With this in mind, raw milk from an individual farm was transformed under controlled conditions at a dairy pilot plant. The production technology included the partial skimming of the evening and morning milk mixture by cream surfacing and the addition of a natural whey starter cultures (NWSC) prepared with four selected Streptococcus thermophilus strains (PON6, PON244, PON261 e PON413). Ten microbial groups were investigated by plate counts from raw milk until ripened cheeses. Lactic acid bacteria (LAB) were in the range 104-105 CFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 106 CFU/g after curd cooking. A rapid pH drop (to 6.05) was registered after almost 3 h from NWSC addition. The levels of members of the Enterobacteriaceae family were at about 103 CFU/ml in raw milk and decreased after curd cooking to 1 log cycle. A similar behavior was shown by the other undesired microbial groups and a complete disappearance of staphylococci was registered. The microbiological counts of 9-month ripened cheeses showed the dominance of LAB and undetectable levels of the undesired bacteria. MiSeq Illumina was applied to better investigate the bacterial composition of ripened cheeses and this technique evidenced that the majority of OTUs belonged to Lactobacillus and Streptococcus genera. The final cheeses were characterized by 67.65% dry matter of which 41.85% of fats and 47.02% of proteins. The main cheese fatty acids were palmitic, oleic and myristic acids and the saturated fatty acids/unsaturated fatty acids ratio was 2.17. Forty-one volatile compounds, including acids, esters, ketones, alcohols, aldehydes, phenols and one terpene were emitted from the cheese. Sensory evaluation showed a general appreciation for the new cheese product by judges.


Subject(s)
Cheese/microbiology , Food Technology/methods , Milk/microbiology , Animals , Cheese/analysis , Fatty Acids/analysis , Female , Lactobacillus/growth & development , Sheep , Streptococcus/growth & development , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism , Volatile Organic Compounds/analysis
16.
Acta Sci Pol Technol Aliment ; 18(2): 153-161, 2019.
Article in English | MEDLINE | ID: mdl-31256543

ABSTRACT

BACKGROUND: Recently, interest in the development of functional foods enriched with bioactive components has increased. Dairy products supplemented with tea extracts known for their health-promoting properties are good examples of such products. However, most of the scientific studies and applications focus on green tea. The present study was established to estimate the effect of Pu-erh tea supplementation on the viability of starter microorganisms and selected physico-chemical and sensory properties of probiotic ABT-yoghurt. METHODS: ABT-yoghurts (Lactobacillus acidophilus La-5, Bifidobacterium animalis ssp. lactis BB-12, Streptococcus thermophilus) were produced from cow’s milk with 0%, 5%, 10% or 15% (v/v) of Pu-erh tea infusion added before the fermentation stage. The products obtained were subjected to the following analyses one day after production (colour profile) and after 7, 14 and 21 days of cold storage: ferric reducing antioxidant power (FRAP) and anti-radical power (ARP) measured against DPPH radical, titratable acidity, pH, texture parameters (back extrusion test), viability of starter cultures and sensory quality (hedonic scale experiment). RESULTS: Pu-erh tea supplementation significantly enhanced the antioxidant potential of probiotic yoghurts as a 3–6.5-fold increase in FRAP and a 10–24-fold increase in ARP values were observed in comparison to plain ABT-yoghurt. Pu-erh tea slightly enhanced the viability of L. acidophilus and reduced the pH of probiotic yoghurts. Higher concentrations of Pu-erh tea caused decreased firmness and consistency while cohesiveness and index of viscosity remained unaffected upon supplementation. The addition of Pu-erh tea infusion modified the colour and sensory properties of the probiotic yoghurts but the sensory quality of the tea yoghurts was rated lower when compared to the plain one. Among all tea yoghurts, the one with 15% Pu-erh tea additive received the highest scores in sensory assessment. CONCLUSIONS: Pu-erh tea may be successfully applied as a functional additive to probiotic yoghurts, signifi- cantly enhancing the antioxidant properties of fermented milk and ensuring a high rate of starter bacteria viability during storage. However, the level of fortification must be carefully chosen as some doses negatively influence texture parameters and sensory quality.


Subject(s)
Antioxidants/pharmacology , Camellia sinensis , Fermentation , Food Handling/methods , Probiotics , Tea , Yogurt/analysis , Animals , Bifidobacterium animalis/growth & development , Cattle , Dietary Supplements , Female , Functional Food , Humans , Hydrogen-Ion Concentration , Lactobacillus acidophilus/growth & development , Microbial Viability , Milk/microbiology , Plant Extracts/pharmacology , Rheology , Streptococcus thermophilus/growth & development , Taste , Viscosity , Yogurt/microbiology
17.
J Microbiol ; 57(9): 769-780, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31201725

ABSTRACT

Understanding global changes of physiological processes at the molecular level during the growth of Streptococcus thermophilus is essential for the rational design of cultivation media and the optimization of bioprocesses. Transcriptomics and proteomics were combined to investigate the global changes at the transcript and protein level during the growth of S. thermophilus. The expression of 1396 genes (FDR ≤ 0.001) and 876 proteins (P < 0.05) changed significantly over time. The most remarkable growth phase dependent changes occurred in the late-lag phase and were related to heterofermentation, glycolysis, peptidoglycan biosynthesis, conversion between amino acids and stress response. The present results could provide theoretical guidance for high-cell-density culture, help design cultivation media, and help attain a high biomass of S. thermophilus.


Subject(s)
Bacterial Proteins/genetics , Streptococcus thermophilus/genetics , Streptococcus thermophilus/metabolism , Transcriptome , Amino Acids/metabolism , Bacterial Proteins/metabolism , Batch Cell Culture Techniques , Fermentation , Gene Expression Profiling , Hydrogen-Ion Concentration , Proteomics , Streptococcus thermophilus/growth & development
18.
Food Chem ; 289: 250-258, 2019 Aug 15.
Article in English | MEDLINE | ID: mdl-30955609

ABSTRACT

Preparation of selenium-enriched probiotics and Se-enrichment of probiotic-fermented blended juices were performed and optimized using orthogonal test. Se content had a significant 13.0-fold increase by the addition of 1% Se-enriched Streptococcus thermophilus starters in juice fermentation. Chemical properties of fermented blended juices were determined. Reducing sugar content decreased significantly after fermentation, and the same downtrend was observed for free amino acids and organic acids, with lactic acid being an exception. Meanwhile, dynamic variation analysis of flavor components during the fermentation, and characteristic aroma-active compounds before and after fermentation were demonstrated by GC-MS and GC-O. Eleven aroma-active substances were identified from juices without fermentation, while 7 characteristic compounds were detected in fermented juices. Furthermore, potential correlations between chemical and flavor characteristics were explored based on multivariate statistical analysis. These results indicate that a potential Se-enriched fermented beverage was established, and the fermentation process led to differences in the chemical substrates and impact odorants.


Subject(s)
Flavoring Agents/analysis , Fruit and Vegetable Juices/analysis , Gas Chromatography-Mass Spectrometry , Selenium/chemistry , Streptococcus thermophilus/growth & development , Amino Acids/analysis , Bioreactors , Citrus sinensis/chemistry , Citrus sinensis/metabolism , Daucus carota/chemistry , Daucus carota/metabolism , Least-Squares Analysis , Malus/chemistry , Malus/metabolism , Odorants/analysis , Principal Component Analysis , Ziziphus/chemistry , Ziziphus/metabolism
19.
J Agric Food Chem ; 67(11): 3198-3208, 2019 Mar 20.
Article in English | MEDLINE | ID: mdl-30838858

ABSTRACT

Electrospun nanofiber membranes are widely investigated in the past few decades as candidates for tissue engineering, which can mimic natural extracellular matrix (ECM) and improve cell adhesion, proliferation, and expression on nanofiber membranes. However, the formation of bacterial biofilms on nanofiber membranes and application of the biofilm-integrated nanofiber membranes remain largely unknown. Here, electrospun cellulose acetate nanofiber membranes are first utilized as scaffold materials for Lactobacillus plantarum ( L. plantarum) biofilm formation. Nanofiber membranes proved to be an excellent scaffold for bacteria biofilm with high stability, where biofilms were interlocked with nanofibers forming a cohesive structure. In comparison with planktonic bacteria, L. plantarum biofilms on nanofiber membranes show excellent gastrointestinal resistance. Instead of decreasing, the number of viable cells increased after 3 h digestion in vitro. The L. plantarum biofilm-integrated nanofiber membranes were used as reusable starter cultures for fermented milk production showing excellent fermentative ability and higher survival of L. plantarum during shelf life. The viable cells in fermented milk remained at 11 log CFU/g throughout the reusable batches, which is far above the required value of 7 log CFU/g in commercial products. In addition, the produced fermented milk possesses shorter fermentation time and higher survival of probiotics during shelf life. The results suggest electrospun nanofiber membranes are ideal scaffold materials for bacteria biofilms immobilization in biotechnology and fermentation engineering, which broaden the potential use of electrospun nanofiber membranes in microbiology and strengthen the application of biofilms in fermentation engineering.


Subject(s)
Food Microbiology/methods , Milk/microbiology , Probiotics/chemistry , Animals , Biofilms , Cattle , Cultured Milk Products/analysis , Cultured Milk Products/microbiology , Fermentation , Food Microbiology/instrumentation , Lactobacillus plantarum/growth & development , Lactobacillus plantarum/metabolism , Nanofibers/chemistry , Streptococcus thermophilus/growth & development , Streptococcus thermophilus/metabolism
20.
Nutrients ; 11(3)2019 Mar 18.
Article in English | MEDLINE | ID: mdl-30889821

ABSTRACT

The current study investigates whether probiotic fermented milk (PFM) and yogurt consumption (YC) are related to both the ingested bacteria taxa and the overall gut microbiota (GM) composition in healthy adults. PFM and YC habits were analyzed in 260 subjects (51% male) by specific questionnaires, and the following groups were considered: (1) PFM groups: nonconsumers (PFM-NC, n = 175) and consumers (PFM, n = 85), divided as follows: Bifidobacterium-containing PFM (Bif-PFM; n = 33), Lactobacillus-containing PFM (Lb-PFM; n = 14), and mixed Bifidobacterium and Lactobacillus-containing PFM (Mixed-PFM; n = 38); (2) PFM-NC were classified as: yogurt nonconsumers (Y-NC; n = 40) and yogurt consumers (n = 135). GM was analyzed through 16S rRNA sequencing. PFM consumers showed higher Bifidobacteria taxa levels compared to NC, from phylum through to species. Specifically, Bif-PFM consumption was related to higher B. animalis levels (p < 0.001), whereas Lb-PFM consumption was associated to higher levels of Bifidobacterium (p < 0.045) and B. longum (p = 0.011). YC was related to higher levels of the yogurt starter Streptococcus thermophilus (p < 0.001). Lactobacilli and the overall GM were not related either to YC or PFM consumption. According to these results, healthy adults might benefit from PFM intake by increasing Bifidobacterium levels.


Subject(s)
Bifidobacterium/growth & development , Gastrointestinal Microbiome , Lactobacillus/growth & development , Milk/microbiology , Probiotics , Streptococcus thermophilus/growth & development , Yogurt/microbiology , Adult , Animals , Female , Fermentation , Humans , Male , RNA, Ribosomal, 16S , Reference Values
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