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Lactic acid bacteria are probiotics in the intestines and have been widely used as natural antioxidants in the food industry. In this study, Enterococcus italicus FM5 with strong antioxidant ability was isolated from fresh milk. The safety evaluation showed that E. italicus FM5 was sensitive to ampicillin, chloramphenicol, erythromycin, vancomycin, rifampicin, and tetracycline, and was not hemolytic. Meanwhile, the whole genome information and biofunctional attributes of this strain were determined and analyzed. Subsequently, E. italicus FM5 was co-cultured with traditional yogurt starters (Streptococcus thermophilus and Lactobacillus bulgaricus) to make fermented milk. The results showed that the addition of E. italicus FM5 could improve the oxygen free radical scavenging ability of the fermented milk, and the scavenging rates of DPPH, ABTS, OH - , and O 2 - radicals reaching up to 95.54 %, 88.35 %, 93.65 %, and 60.29 %, respectively. Furthermore, the addition of E. italicus FM5 reduced the curd time and improved the water holding capacity of the fermented milk. Besides, the growth of Lb. bulgaricus was significantly promoted when co-cultured with E. italicus FM5, thus the survival cells were increased compared with the traditional fermentation processes. Therefore, this study emphasized the potential to manufacture fermented milk by the co-cultivation of E. italicus with traditional yogurt starters., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

For complete utilization of high glucose at ∼100 g/L, a high cell density (HCD) continuous fermentation system was established using Lb. delbrueckii NCIM 2025 for the bioproduction of lactic acid (LA). An integrated membrane cell recycling system coupled with the continuous bioreactor, aided to achieve the highest 34.77 g/L h LA productivity and 0.94-0.98 g/g yield. ∼34 times higher productivity was observed (in comparison to batch fermentation conducted in this study), when the continuous operations were carried out at the maximum dilution rate and wet cell weight i.e. 0.36 h -1 and 230 g/L, respectively. These results show the potential of this method for large-scale lactic acid production because it not only produces high titers but also ensures that glucose is used effectively. The method's superior performance in comparison to earlier studies suggests it as an affordable and sustainable alternative for the production of LA.

Direct molecular methods such as real-time polymerase chain reaction (qPCR) and propidium monoazide (PMA)-qPCR have been successfully used for quantifying viable microorganisms in the food industry. This study attempted to use qPCR and PMA-qPCR for quantifying Lactobacillus delbrueckii spp. bulgaricus sp1.1 physiological states. The qPCR standards of the 16S rRNA gene were employed to calibrate the qPCR assay, which contributed to an amplification efficiency of 98.42%. The number of copies of the 16S rRNA gene was linearly related to cell density, and this linear relationship was used to construct a quantitative curve ( R 2 =0.9981) with a detection limit of 15.1 colony-forming units mL -1 ·reaction -1 . qPCR in combination with an optimal PMA concentration (60 μM) helped in discriminating and quantifying the viable cells, without any interference by heat-killed cells. Compared with the conventional methods, the population heterogeneity of viable, culturable, dormant-like and membrane-permeabilized cells were well identified and quantified using qPCR during L . delbrueckii spp. bulgaricus sp1.1 batch culture. Despite the restriction in the enumeration of lysed cells, qPCR-based methods facilitated reliable identification and quantification of bacterial physiological states and provided additional knowledge on the dynamics of L . delbrueckii spp. bulgaricus sp1.1 physiological states.

The xyloglucanase gene (RmXEG12A) from Rhizomucor miehei CAU432 was successfully expressed in Pichia pastoris. The highest xyloglucanase activity of 25,700 U mL -1 was secreted using high cell density fermentation. RmXEG12A was optimally active at pH 7.0 and 65 °C, respectively. The xyloglucanase exhibited the highest specific activity towards xyloglucan (7915.5 U mg -1 ). RmXEG12A was subjected to hydrolyze tamarind powder to produce xyloglucan oligosaccharides with the degree of polymerization (DP) 7-9. The hydrolysis ratio of xyloglucan in tamarind powder was 89.8%. Moreover, xyloglucan oligosaccharides (2.0%, w/w) improved the water holding capacity (WHC) of yoghurt by 1.1-fold and promoted the growth of Lactobacillus bulgaricus and Streptococcus thermophiles by 2.3 and 1.6-fold, respectively. Therefore, a suitable xyloglucanase for tamarind powder hydrolysis was expressed in P. pastoris at high level and xyloglucan oligosaccharides improved the quality of yoghurt., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Lactobacillus delbrueckii JCM 1002 T grows on highly polymerized inulin-type fructans as its sole carbon source. When it was grown on inulin, a > 10 kb long gene cluster inuABCDEF (Ldb1381-1386) encoding a plausible ABC transporter was suggested to be induced, since a transcriptome analysis revealed that the fourth gene inuD (Ldb1384) was up-regulated most prominently. Although Bacillus subtilis 168 is originally unable to utilize inulin, it became to grow on inulin upon heterologous expression of inuABCDEF. When freshly cultured cells of the recombinant B. subtilis were then densely suspended in buffer containing inulin polymers and incubated, inulin gradually disappeared from the buffer and accumulated in the cells without being degraded, whereas levan-type fructans did not disappear. The results imply that inuABCDEF might encode a novel ABC transporter in L. delbrueckii to "monopolize" inulin polymers selectively, thereby, providing a possible advantage in competition with other concomitant inulin-utilizing bacteria., (© 2021. The Author(s).)

Gene feoA plays an important role in cell growth because of its function of transport Fe 2+ which is a necessary element for cells. In this study, the recombinant plasmid pUC19-feoA-Tet was successfully constructed using the inserted gene inactivation method. Using the homologous recombination technique, the tet gene was used as a resistance screening marker to knock out the feoA gene of Lactobacillus delbrueckii subsp. bulgaricus 34.5 (strain 34.5). Comparative analysis of growth curves revealed the growth changes in the absence of feoA gene in strain 34.5. The results showed that the growth of the bacteria was prolonged by 2 h and could be restored in the stationary phase. To further study whether feoA is related to the cell division of strain 34.5, the qPCR experiment was carried out. The results showed that, compared with the wild-type strain, the expression of genes related to cell division in the mutant strain was up-regulated in the pre-log phase, down-regulated in the late-log phase, and returned to the original level in the stationary phase. These findings provide ideas for Lactobacillus delbrueckii subsp. bulgaricus to control division and cell cycle., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The application of Streptococcus thermophilus S-3 into yogurt production was studied and the structural properties of the generated exopolysaccharides (EPS-S3) were characterized. The proposed structure of EPS-S3 was obtained. EPS-S3 contained a high ratio of N-Acetyl-galactosamine with the Mw of 574 kDa, which was higher than that of AR333 (314 kD) leading to higher apparent viscosity. Streptococcus thermophilus strain S-3 was co-cultured with Lactobacillus delbrueckii for yogut production which highly increased the acidifying rate and post-acidification rate. The quality of the co-cultured yogurts in terms of apparent viscosity, syneresis capacity, water holding capacity and rheological properties were much better than that by using Lactobacillus bulgaricus only. The production mechanism of EPS-S3 from gene regulated level was also discussed which is helpful to facilitate the application of Streptococcus thermophilus strain into milk production., Competing Interests: Declaration of competing interest The authors declare no conflict of interests., (Copyright © 2021. Published by Elsevier B.V.)

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 β-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., (Copyright © 2020. Published by Elsevier Ltd.)

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., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Several species of eukaryotic organisms living in the high mountain areas of Armenia with naturally occurring levels of radiation have high adaptive responses to radiation. We speculate on the role of the gastrointestinal microbiota in this protection against radiation. Therefore, seventeen microorganisms with high antagonistic activities against several multi-drug-resistant pathogens were isolated from the human and animal gut microbiota, as well as from traditional Armenian fermented products. These strains were tested in vivo on Wistar rats to determine their ability to protect the eukaryotic host against radiation damages. The efficiency of the probiotics' application and the dependence on pre- and post-radiation nutrition of rats were described. The effects of Lactobacillus rhamnosus Vahe, isolated from a healthy breastfed infant, and Lactobacillus delbrueckii IAHAHI, isolated from the fermented dairy product matsuni, on the survival of irradiated rats, and their blood leucocyte and glucose levels, were considered to be the most promising, based on this study's results.

Objectives: To produce nattokinase in a food-grade expression system and evaluate its thrombolytic activity in vitro., Results: No nattokinase activity from reconstituted strains was observed in simulated gastric juice, but the enzyme was stable in intestinal fluid, the relative activity of which was found to be 60% after 4 h. Due to the nattokinase being produced intracellularly by recombinant bacterial strains, the persistence of the bacteria in gastric juice ensured transmission of the nattokinase into intestinal juice. Because of subsequent disintegration of the bacteria, the highest nattokinase activity was observed after 3 h at approximately 32%, following its carriage within the recombinant strains to the intestinal fluid., Conclusions: This study demonstrated that nattokinase from recombinant strains exhibited good thrombolytic activity in vitro and may be used by the dairy fermentation industry for the development of novel thrombolytic functional foods.

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 D 2 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 10 6 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., (© 2020 Institute of Food Technologists®.)

Bergamot essential oil (BEO) is well-known for its food preservation activity, as well as anticancer efficacy. However, the poor BEO water solubility and deriving low bioaccessibility have limited its wider applications. The incorporation in nanoemulsions of BEO and its refined fractions was investigated to enhance its dispersibility in water to promote its antimicrobial activity, tested against Escherichia coli, Lactobacillus delbrueckii, and Saccharomyces cerevisiae, and its cytotoxicity already at low concentrations. Different nanoemulsion formulations were tested based on food-grade ingredients, which were characterized in terms of hydrodynamic diameter and polydispersity index, and physical stability. The antimicrobial activity against all the tested micro-organisms was observed to be higher for BEO in its initial composition, than the light fraction, richer in d-limonene, ß-pinene, and γ-terpinene, or the heavy fraction, richer in linalyl acetate and linalool. Remarkably, the use of BEO nanoemulsions notably enhanced the antimicrobial activity for all the tested oils. BEO exhibited also a measurable cytotoxic activity against Caco-2 cells, which was also enhanced by the use of the different nanoemulsions tested, in comparison with free oil, which discourages the direct use of BEO nanoemulsions as a food preservative. Conversely, BEO nanoemulsions might find use in therapeutic applications as anticarcinogenic agents.

To decrease d-lactic acid production cost, sugarcane molasses and soybean meal, low-cost agro-industrial wastes, were selected as feedstock. First, sugarcane molasses was used directly by Lactobacillus delbrueckii S-NL31, and the nutrients were released from soybean meal by protease hydrolysis. Subsequently, to ensure intensive substrate utilization and enhanced d-lactic acid production from sugarcane molasses and soybean meal, adaptation of L. delbrueckii S-NL31 to substrates was performed through adaptive laboratory evolution. After two-phase adaptive laboratory evolution, the evolved strain L. delbrueckii S-NL31-CM3-SBM with improved cell growth and d-lactic acid production on sugarcane molasses and soybean meal was obtained. To decipher the potential reasons for improved fermentation performance, a metabolomics-based approach was developed to profile the differences of intracellular metabolism between initial and evolved strain. The in-depth analysis elucidated how the key factors exerted influence on d-lactic acid biosynthesis. The results revealed that the enhancement of glycolysis pathway and cofactor supply was directly associated with increased lactic acid production, and the reinforcement of pentose phosphate pathway, amino acid metabolism, and oleic acid uptake improved cell survival and growth. These might be the main reasons for significantly improved d-lactic acid production by adaptive laboratory evolution. Finally, fed-batch simultaneous enzymatic hydrolysis of soybean meal and fermentation process by evolved strain resulted in d-lactic acid levels of 112.3 g/L, with an average production efficiency of 2.4 g/(L × h), a yield of 0.98 g/g sugar, and optical purity of 99.6%. The results show the applicability of d-lactic acid production in L. delbrueckii fed on agro-industrial wastes through adaptive laboratory evolution.

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., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Purpose: The goal of this paper is to describe the green conversion of agricultural waste products, such as molasses and corn steep liquor, into large amounts of D(-) lactic acid using a facilitated multipulse fed-batch strategy and affordable pH neutralizer. This is a very low-cost process because there is no need for hydrolysis of the waste products. The fed-batch strategy increases lactic acid productivity by avoiding inhibition caused by a high initial substrate concentration, and the selected controlling agent prevents cell stress that could be caused by high osmotic pressure of the culture media., Methods: The effects of different carbon and nitrogen sources on lactic acid production were investigated, and the best concentrations of the medium components were determined. To optimize the culture conditions of the Lactobacillus delbrueckii strain, the effects of pH control, temperature, neutralizing agent, agitation, and inoculum size in batch cultures were investigated. Fed-batch strategies were also studied to improve production and productivity., Result: A high titer of D(-) lactic acid (162g/liter) was achieved after 48 hours of fermentation. Productivity at this point was 3.37 g/L·h. The optimum conditions were a temperature of 39°C, pH 5.5 controlled by the addition of Ca(OH) 2 , agitation at 150 rpm, and inoculum size of 25% ( v / v )., Conclusion: The production of high optical purity D(-) lactic acid through L . delbrueckii fermentation with molasses and corn steep liquor is a promising economical alternative process that can be performed on the industrial scale., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Susan Michelz Beitel et al.)

Cell growth of lactic acid bacteria is of vital importance in starter culture manufacture in the dairy industry. However, one of the major stumbling blocks in the understanding of bacterial cell growth relates to challenges in the quantification of the cell division or death. To overcome these shortcomings, the intracellular fluorescent cell tracking assay was implemented to monitor Lactobacillus delbrueckii subsp. bulgaricus sp1.1 cell division and death. Optimization of fluorescent dye concentration suggested it could be applied in the tracking of cell division without cytotoxicity. Technical validation of the fluorescent tracking demonstrated this assay was accurate for quantitatively analyzing cell division. Furthermore, the cell death was quantified using the precursor cohort distribution of the time-series fluorescence data in batch culture. The results indicated a dynamic and unbalanced relationship between bacterial cell division and death after exponential phase in the batch culture. These findings suggested that fluorescent dye tracking is a powerful tool for monitoring L. bulgaricus sp1.1 cell division and death and can provide valuable information for bacterial growth behavior in batch culture., (Copyright © 2020. Published by Elsevier B.V.)

Siraitia grosvenorii fruit (SGF) has been used as a natural sweetener and traditional medicine in China for more than two centuries. This study evaluated the effect of SGF extract supplementation (0.5%, 1%, and 2%) on the chemical, microbial and sensory properties of probiotic yogurt. The antioxidant, angiotensin-converting-enzyme inhibitory (ACE-I) and antibacterial bioactivities were determined. SGF extract supplementation improved some of the chemical and physicochemical characteristics. Probiotic yogurt with the fruit extract had significantly more Lactobacillus casei and Lactobacillus bulgaricus, whereas there was no significant effect on the number of Streptococcus thermophiles. The bioactivities were significantly increased by SGF extract supplementation. Probiotic yogurt with 2% SGF extract showed the highest antioxidant, ACE-I, and antibacterial activities, whereas the one with 1% SGF extract conferred the highest sensory attributes score. Overall, SGF extract offers a promising option as a dietary supplement to produce novel dairy products that have high nutritional and bioactivity values., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background and Objective: Hepatorenal syndrome (HRS) is a major public health problem in which both liver and kidney dysfunctions are encountered. The present research aimed to investigate the beneficial use of micro-encapsulated probiotic alone (Bifidobacterium bifidum, Lactobacillus delbrueckii and Streptococcus thermophilus mixture) or with green tea alcohol extract in HRS model in rats., Materials and Methods: Flavonoids content and in vitro antioxidant activity of the extract were assessed. The animal experiment consisted of 4 groups; control healthy, control with HRS and two test groups with HRS and treated with either the encapsulated probiotic mixture alone or with green tea extract. After 3 weeks; urinary creatinine was determined in 24 h rat urine samples. Colonic microbiota was assessed in faeces. Plasma malondialdehyde, nitrite, C-reactive protein, creatinine, uric acid, urea and the activity of transaminases, catalase (CAT) and angiotensin-1 converting enzyme (ACE-1) were determined with calculation of creatinine clearance., Results: Results showed significant increase in all biochemical parameters of HRS control except for ACE-1, CAT and creatinine clearance that experienced significant reduction along with dysbiosis compared to healthy control. Test groups showed improvement in all biochemical parameters with superiority to probiotic-green tea extract combination. Both treatments produced significant increase in fecal B. bifidum, S. thermophilus and L. bulgaricus and reduction of Staphylococci and Coliform. The effect of probiotic-green tea extract combination was more pronounced concerning the last three. Flavonoids and antioxidant activity of the extract were 1.325±0.01 mg quercetin/g and 98±1.66%, respectively., Conclusion: Administration of micro-encapsulated probiotic with or without alcohol green tea extract exerted significant prevention of HRS in rat with superiority to probiotic-green tea extract combination.

Modified reinforced clostridial medium (mRCM) was developed and evaluated for the differential enumeration of Lactobacillus delbrueckii ssp. bulgaricus. Lactobacillus bulgaricus, an important species of lactic acid bacteria with health benefits, is used in the production of yogurt and other fermented foods. Our results showed that supplementing reinforced clostridial medium with 0.025% CaCl 2 , 0.01% uracil, and 0.2% Tween 80 (mRCM) significantly enhanced the growth rate of L. bulgaricus RR and ATCC 11842 strains as measured by the optical densities of these strains after 12 h of incubation at 42°C. The bacterial populations (plate count) of the RR and ATCC 11842 strains were 0.76 and 0.77 log cfu/g higher in mRCM than in de Man, Rogosa, and Sharpe and reinforced clostridial medium media, respectively. Conversely, the population counts for other bacterial species (Bifidobacterium, Lactobacillus rhamnosus, and Lactobacillus reuteri) were significantly inhibited in the mRCM medium. The addition of aniline blue dye to mRCM (mRCM-blue) improved the selectivity of L. bulgaricus in mixed lactic bacterial cultures compared with de Man, Rogosa, and Sharpe medium and lactic agar with regard to colony appearance and morphology. The mRCM-blue performed better than the conventional medium in culturing, enumerating, and differentiating L. bulgaricus. Therefore, mRCM-blue could be used as a selective medium to enhance the growth and differentiation of L. bulgaricus in order to meet the increasing demand for this beneficial species of bacteria., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The contribution of two starter (Lactobacillus delbrueckii and Streptococcus thermophilus) and nine non-starter (Enterococcus casselliflavus, Enterococcus faecalis, Enterococcus durans, Enterococcus gallinarum, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, Pediococcus acidilactici and Pediococcus pentosaceus) species of lactic acid bacteria (LAB) to the volatile organic compounds (VOCs) of Caciocavallo Palermitano cheese was investigated. The strains used in this study were isolated during the production/ripening of the stretched cheese and tested in a cheese-based medium (CBM). The fermented substrates were analyzed for the growth of the single strains and subjected to the head space solid phase micro-extraction (HS-SPME) and gas chromatography - mass spectrometry (GC-MS). The 11 strains tested were all able to increase their numbers in CBM, even though the development of the starter LAB was quite limited. GC-MS analysis registered 43 compounds including seven chemical classes. A lower diversity of VOCs was registered for the unfermented curd based medium (CuBM) analyzed for comparison. The class of ketones represented a consistent percentage of the VOCs for almost all LAB, followed by alcohols and esters. The volatile profile of Pediococcus acidilactici and Lactobacillus delbrueckii was mainly characterized by 2-butanol, butanoic acid and hexanoic acid and their esters, while that of Lactobacillus casei and Lactobacillus rhamnosus was characterized by 2,3-butanedione and 2-butanone, 3-hydroxy. In order to correlate the VOCs produced by Caciocavallo Palermitano cheeses with those generated by individual LAB, the 4-month ripened cheeses resulting from the dairy process monitored during the isolation of LAB were also analyzed for the volatile chemical fraction and the compounds in common were subjected to a multivariate statistical analysis. The canonical analysis indicated that the VOCs of the ripened cheeses were mainly influenced by E. gallinarum, L. paracasei, L. delbrueckii, L. rhamnosus and L. casei and that 1-hexanol, o-xylene and m-xylene were the cheese VOCs highly correlated with LAB., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In a screening for proteolytically active lactic acid bacteria, three strains, Lactobacillus delbrueckii ssp. lactis 92202, Lactobacillus helveticus 92201, and Lactobacillus delbrueckii ssp. bulgaricus 92059, showed the highest activities following growth in milk. All three strains degraded α- and β-casein, but did not hydrolyse κ-casein. HPLC analysis of skim milk fermentation revealed increasing amounts of peptides after 5 and 10 h with Lb. d. ssp. bulgaricus 92059. Hydrolysates obtained with Lb. d. ssp. lactis 92202 and Lb. d. ssp. bulgaricus 92059 revealed the highest angiotensin-converting enzyme-inhibitory effect. The effect was dose dependent. Almost no effect (<10%) was seen for Lb. helveticus 92201. For Lb. d. ssp. bulgaricus 92059, maximal inhibition of approx. 65% was reached after 25 h of fermentation. In an in vitro assay measuring potential immunomodulation, hydrolysates of the three strains yielded anti-inflammatory activities in the presence of TNF-α. However, the effects were more pronounced at lower hydrolysate concentrations. In the absence of TNF-α, slight pro-inflammatory effects were observed. The hydrolysate of Lb. d. ssp. bulgaricus 92059, when purified by means of solid-phase extraction, exhibited pro-inflammatory activity. Sour whey containing Lb. d. ssp. bulgaricus 92059 cells showed pro-inflammatory activity while cell-free sour whey was clearly anti-inflammatory. In the purified hydrolysate, 20 different α- and β-casein (CN)-derived peptides could be identified by LC-MS. Most peptides originated from the central and C-terminal regions of β-casein. Peptide length was between 9 (β-CN(f 59-67)) and 22 amino acids (β-CN(f 117-138)).

Background: The efficacy of Lactobacillus delbrueckii subsp. bulgaricus as starter cultures for the dairy industry depends largely on the number of viable and active cells. Freeze-drying is the most convenient and successful method to preserve the bacterial cells. However, not all strains survived during freeze-drying., Methods: The effects of six substances including NaCl, sorbitol, mannitol, mannose, sodium glutamate, betaine added to the MRS medium on the growth and freeze-drying survival rate and viable counts of Lb. delbrueckii subsp. bulgaricus were studied through a single-factor test and Plackett-Burman design. Subsequently, the optimum freeze-drying conditions of Lb. delbrueckii subsp. bulgaricus were determined., Results: Lb. delbrueckii subsp. bulgaricus survival rates were up to the maximum of 42.7%, 45.4%, 23.6%, while the concentrations of NaCl, sorbitol, sodium glutamate were 0.6%, 0.15%, 0.09%, respectively. In the optimum concentration, the viable counts in broth is 6.1, 6.9, 5.13 (&times;108 CFU/mL), respectively; the viable counts in freeze-drying power are 3.09, 5.2, 2.7 (&times;1010 CFU/g), respectively., Conclusions: Three antifreeze factors including NaCl, sorbitol, sodium glutamate have a positive effect on the growth and freeze-drying of Lb. delbrueckii subsp. bulgaricus. The results are beneficial for developing Lb. delbrueckii subsp. bulgaricus.

The effect of W 1 /O/W 2 emulsion incorporation in set-type yogurt on the acidification process, physicochemical properties, bacterial growth kinetics and structural characteristics was investigated. The W 1 /O/W 2 emulsion was formed by using a two-step homogenisation process and milk as the W 1 and W 2 phases, and stability was monitored with optical microscopy and cryo-SEM. Adding the W 1 /O/W 2 emulsions reduced the acidification rate, viscosity and water retention capacity. Texture (adhesiveness, cohesiveness, hardness, and gumminess) differed in yogurts containing W 1 /O/W 2 emulsion compared to controls during the acidification process, however, trends became stable during storage. The growth of S. thermophilus during the acidification process of yogurt was reduced in the presence of W 1 /O/W 2 emulsion while L. bulgaricus trended higher during storage. This study shows that yogurts containing W 1 /O/W 2 emulsion are feasible subject to processing modification., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Yogurt products fermented with probiotic bacteria are a consumer trend and a challenge for functional food development. So far, limited research has focused on the behavior of the various probiotic strains used in milk fermentation. In the present study, we characterized folic acid production and the sensory and textural characteristics of yogurt products fermented with probiotic bacteria. Yogurt fermented with Lactobacillus plantarum had improved nutrient content and sensory and textural characteristics, but the presence of L. plantarum significantly impaired the growth and survival of Lactobacillus delbrueckii ssp. bulgaricus during refrigerated storage. Overall, L. plantarum was a good candidate for probiotic yogurt fermentation; further studies are needed to understand the major metabolite path of lactic acid bacteria in complex fermentation., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in the yogurt consortium enhances the growth rate and size of each population. In contrast, the independent growth of the two species in milk leads to a slower growth rate and a smaller population size. In this study, we report the first evidence that the urease activity of S. thermophilus increases the intracellular pH of L. delbrueckii in the absence of carbon source. However, in milk, in the presence of lactose the alkalizing effect of urea-derived ammonia was not detectable. Nevertheless, based on glucose consumption and lactic acid production at different pH in , L. delbrueckii showed an optimum of glycolysis and homolactic fermentation at alkaline pH values. In milk, we observed that ammonia provided by urea hydrolysis boosted lactic acid production in S. thermophilus and in L. delbrueckii when the species were grown alone or in combination. Therefore, we propose that urease activity acts as an altruistic cooperative trait, which is costly for urease-positive individuals but provides a local benefit because other individuals can take advantage of urease-dependent ammonia release., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Bovine lactoferrin (bLf) is a natural iron-binding protein and it has been suggested to be a prebiotic agent, but this finding remains inconclusive. This study explores the prebiotic potential of bLf in 14 probiotics. Initially, bLf (1-32 mg/mL) treatment showed occasional and slight prebiotic activity in several probiotics only during the late experimental period (48, 78 h) at 37 °C. We subsequently supposed that bLf exerts stronger prebiotic effects when probiotic growth has been temperately retarded. Therefore, we incubated the probiotics at different temperatures, namely 37 °C, 28 °C, room temperature (approximately 22-24 °C), and 22 °C, to retard or inhibit their growth. As expected, bLf showed more favorable prebiotic activity in several probiotics when their growth was partially retarded at room temperature. Furthermore, at 22 °C, the growth of Bifidobacterium breve, Lactobacillus coryniformis, L. delbrueckii, L. acidophilus, B. angulatum, B. catenulatum, and L. paraplantarum were completely blocked. Notably, these probiotics started regrowing in the presence of bLf (1-32 mg/mL) in a significant and dose-dependent manner. Accordingly, bLf significantly increased the growth of Pediococcus pentosaceus, L. rhamnosus, and L. paracasei (BCRC 17483; a locally isolated strain) when their growth was retarded by incubation at 22 °C. In conclusion, bLf showed inconsistent prebiotic activity in the 14 probiotics at 37 °C, but revealed strong prebiotic activity in 10 probiotic strains at 22 °C. Therefore, this study enables determining additional roles of Lf in probiotic strains, which can facilitate developing novel combinational approaches by simultaneously using Lf and specific probiotics.

Inulin is a readily available feedstock for cost-effective production of biochemicals. To date, several studies have explored the production of bioethanol, high-fructose syrup and fructooligosaccharide, but there are no studies regarding the production of D-lactic acid using inulin as a carbon source. In the present study, chicory-derived inulin was used for D-lactic acid biosynthesis by Lactobacillus bulgaricus CGMCC 1.6970. Compared with separate hydrolysis and fermentation processes, simultaneous saccharification and fermentation (SSF) has demonstrated the best performance of D-lactic acid production. Because it prevents fructose inhibition and promotes the complete hydrolysis of inulin, the highest D-lactic acid concentration (123.6 ± 0.9 g/L) with a yield of 97.9 % was obtained from 120 g/L inulin by SSF. Moreover, SSF by L. bulgaricus CGMCC 1.6970 offered another distinct advantage with respect to the higher optical purity of D-lactic acid (>99.9 %) and reduced number of residual sugars. The excellent performance of D-lactic acid production from inulin by SSF represents a high-yield method for D-lactic acid production from non-food grains.

Background: White mold-ripened cheeses were investigated with the objective of proposing a colorimetric method to monitor the surface growth of Penicillium candidum and to evaluate the influence of the mesophilic (homofermentative (QMO) and heterofermentative (QMLD)) and thermophilic (QT) starter cultures on the physicochemical composition and sensory description., Results: The whiteness index was effective in proving the appearance of superficial mycelium and the stability of white mold growth. The lactic cultures showed significant influence on most of the physicochemical analyses. The cheese made with thermophilic lactic culture had a 1 day gain in the growth of mycelium on the surface; nevertheless, the appearance of this product was potentially not acceptable for consumers. The heterofermentative mesophilic cheese had a better appearance and texture profile. However, the homofermentative mesophilic cheese showed aspects of fresh cheese and was acceptable for a wide range of consumers., Conclusion: The whiteness index was efficient to monitor the surface growth of P. candidum. The highest proteolytic effect was found in the QMLD and QT cultures. However, the cheese elaborated with the QMLD culture showed the best sensory acceptance. © 2015 Society of Chemical Industry., (© 2015 Society of Chemical Industry.)

The objective of this study was to investigate the effect of 2 plant leaf extracts on fermentation mechanisms and health-promoting activities and their potential as a nutraceutical prebiotics ingredient for application in dairy products. The individual active phenolic compounds in the plant extract-supplemented milk and yogurts were also identified. Compared with control fermentation, the plant extracts significantly increased the growth and acidification rate of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. In particular, plant extract components, including monosaccharides, formic acid, and hydroxycinnamic acid, such as neo-chlorogenic, chlorogenic, and caffeic acid, together play a stimulatory role and cause this beneficial effect on the growth of yogurt culture bacteria through fermentation. In addition, supplementation with the plant extracts enhanced antioxidant activities with increased total phenolic contents, especially the highest antioxidant activity was observed in yogurt supplemented with Cudrania tricuspidata leaf extract., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The diverse and dynamic microbiota of the gastrointestinal tract represents a vast source of bioactive substances. These include bacteriocins, which are antimicrobial peptides with the potential to modulate gut populations to impact positively on human health. Although several gut-derived bacteriocins have been isolated, there remain only a few exceptional studies in which their influence on microbial populations within the gut has been investigated. To facilitate such investigations, in vitro faecal fermentation systems can be used to simulate the anaerobic environment of the colon. In this instance, such a system was employed to explore the impact of bactofencin A, a novel broad spectrum class IId bacteriocin produced by gut isolates of Lactobacillus salivarius, on intestinal populations and overall microbial diversity. The study reveals that, although bactofencin A is a broad spectrum bacteriocin, it has a relatively subtle influence on intestinal communities, with a potentially positive impact on anaerobic populations such as Bacteroides, Clostridium and Bifidibacterium spp. The strategy taken is an important first step in investigating the merits of using bactofencin A to manipulate the gut microbiota in a beneficial way for health., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The catabolite control protein A (CcpA) is a kind of multi-effect regulatory protein. In the study, the effect of the inactivation of CcpA and aerobic conditions on the growth, metabolic production, and stress tolerance to heat, oxidative, and cold stresses in Lactobacillus delbrueckii ssp. bulgaricus was investigated. Results showed that inactivation of CcpA distinctly hindered growth. Total lactic acid concentration was significantly lower in aerobiosis for both strains and was lower for the mutant strain than L. bulgaricus. Acetic acid production from the mutant strain was higher than L. bulgaricus in aerobiosis compared with anaerobiosis. Enzyme activities, lactate dehydrogenase (LDH), phosphate fructose kinase (PFK), pyruvate kinase (PK), and pyruvic dehydrogenase (PDH), were significantly lower in the mutant strain than L. bulgaricus. The diameters of inhibition zone were 13.59 ± 0.02 mm and 9.76 ± 0.02 mm for L. bulgaricus in anaerobiosis and aerobiosis, respectively; and 8.12 ± 0.02 mm and 7.38 ± 0.02 mm for the mutant in anaerobiosis and aerobiosis, respectively. For both strains, cells grown under aerobic environment possess more stress tolerance. This is the first study in which the CcpA-negative mutant of L. bulgaricus is constructed and the effect of aerobic growth on stress tolerance of L. bulgaricus is evaluated. Although aerobic cultivation does not significantly improve growth, it does improve stress tolerance., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: The intake of fermented milk products, especially yoghurts, has been systematically increasing for a few decades. The purpose of this work was to obtain milk products fermented with a mix of bacterial cultures (yoghurt bacteria and Lactobacillus acidophillus LA-5) and enriched with selected milk protein preparations. Secondly, the aim of the work was to determine physiochemical and rheological properties of the obtained products., Methods: The following additives were applied in the experiment: whey protein concentrate (WPC 65), whey protein isolate (WPI), demineralised whey powder (SPD), caseinoglycomacropeptide (CGMP), α-lactalbumin (α-la), sodium caseinate (KNa) and calcium caseinate (KCa). Milk was fermented using probiotic strain Lactobacillus acidophillus LA-5 and a typical yoghurt culture. The products were analysed in terms of the survivability of bacterial cells during refrigerated storage, rheological properties and syneresis. Fermented milk products were obtained using blends of bacterial strains: ST-B01:Lb-12 (1:1), ST-B01:Lb-12:LA-5 (1:1:2)., Results: Milk beverages fermented with typical yoghurt bacteria and LA-5 strain showed intensive syneresis. The addition of LA-5 strain caused formation of harder acid gels, comparing to typical yoghurts. Milk products which were prepared from skimmed milk possessed higher values of hardness and consistency coefficient. The increase of concentrations of milk preparations (except of WPI) did not cause significant differences in the hardness of acidic gels obtained by fermentation of mixed culture with a probiotic strain., Conclusions: The applied preparations improved physiochemical properties of the milk beverages which were prepared with a probiotic strain. The increase of protein milk preparations concentration resulted in a gradual decrease of the secreted whey. Among the products that were made of full milk powder and were subjected to three weeks of refrigerated storage the highest survivability of Lb. acidophilus LA-5 was noticed in the samples fortified with 1% WPC.

Background: Although several studies have found probiotics encouraging in prevention of gestational diabetes mellitus (GDM), the evidence for the use of probiotics in diagnosed GDM is largely limited. The aim of this study was to assess the effect of a probiotic supplement capsule containing four bacterial strains on glucose metabolism indices and weight changes in women with newly diagnosed GDM., Methods: Sixty-four pregnant women with GDM were enrolled into a double-blind placebo-controlled randomized clinical trial. They were randomly assigned to receive either a probiotic or placebo capsule along with dietary advice for eight consecutive weeks. The trend of weight gain along with glucose metabolism indices was assayed., Results: During the first 6 weeks of the study, the weight gain trend was similar between the groups. However, in the last 2 weeks of the study, the weight gain in the probiotic group was significantly lower than in the placebo group (p < 0.05). Fasting blood sugar (FBS) decreased in both intervention (from 103.7 to 88.4 mg/dl) and control (from 100.9 to 93.6 mg/dl) groups significantly, and the decrease in the probiotic group was significantly higher than in the placebo group (p < 0.05). Insulin resistance index in the probiotic group had 6.74% reduction over the study period (p < 0.05). In the placebo group, however, there was an increase in insulin resistance index (6.45%), but the observed change in insulin resistance was not statistically significant. Insulin sensitivity index was increased in both groups. The post-intervention insulin sensitivity index in the probiotic group was not significantly different from placebo when adjusted for the baseline levels., Conclusions: The probiotic supplement appeared to affect glucose metabolism and weight gain among pregnant women with GDM. This needs to be confirmed in other settings before a therapeutic value could be approved.

Background: Goat milk production in Shaanxi province is dominant in China, but the product is mainly infant formula and adult milk powder; product homogeneity is serious and has no goat yoghurt with probiotic culture., Methods: The effect of bacteria proportion (1:3:1, 1:2:1, 1:1:1, 2:1:1, 3:1:1) on pH, acidity, and viable counts and sensory evaluation of goat milk fermented by probiotics including L. acidophilus, B. bifidum  or L. casei besides, S. thermophilus and L. bulgaricus for developing AB-goat yoghurt and BC-goat yoghurt was investigated., Results: The optimum bacteria proportion of L. acidophilus : B. bifidum : S. thermophilus and L. bulgaricus for AB-goat yoghurt and B. bifidum : L. casei : S. thermophilus and L. bulgaricus for BC-goat yoghurt were both 2:1:1. The pH, acidity, the viable counts of L. acidophilus and B. bifidum, the total viable counts were respectively 4.60, 7.73 (g/L), 3.50×107 cfu/mL, 3.40×107 cfu/mL and 2.30×109 cfu/mL in AB-goat yoghurt. The pH, acidity, the viable counts of B. bifidum and L. casei, the total viable counts were respectively  4.61, 8.16 (g/L), 7.60×107 cfu/mL, 5.60×107 cfu/mL and 2.04×109 cfu/mL in BC-goat yoghurt., Conclusions: The bacteria proportion had a significant effect on fermentation of AB- and BC-goat yoghurt, the results are beneficial for developing AB-goat yoghurt and BC-goat yoghurt.

Background: Lactobacillus bulgaricus LB6 is a bacterium which was selected in the commercial yoghurt with high angiotensin converting enzyme (ACE) inhibitory activity. Preparation of concentrated starter cultures via freeze drying is of practical importance to dairy and food industries., Methods: We optimized the optimal sugar alcohol and proteins for Lactobacillus bulgaricus LB6 during the process of freeze drying using a Plackett-Burman design. In our initial tests survival rate and the number of viable cells were associated with the type of lyoprotectant used and so our optimization protocol focused on increasing survival rate. Substances that had previously had a protective effect during freeze drying were investigated, for example: mannitol, sorbitol, xylitol, meso-erythritol, lactitol, whey protein isolate 90, bovine serum albumin, and whey protein concentrate 80 and soy protein isolate 70., Results: We found that the optimum sugar alcohol and proteins for survival of Lactobacillus bulgaricus LB6 were whey protein concentrate (p = 0.0040 for survival rate), xylitol (p = 0.0067 for survival rate) and sorbitol (p = 0.0073 for survival rate), they showed positive effect (whey protein concentrate and sorbitol) or negative effect (xylitol)., Conclusions: The effectiveness of three chosen sugar alcohols and protein implied that they could be used as lyoprotectant for Lactobacillus bulgaricus LB6 in the further research, the optimal composition of sugar alcohol and protein for the lyoprotectant use must be established.

Limited availability of resources and increased amounts of waste coupled with an ever-increasing demand for raw materials are typical characteristics of our times. As such, there is an urgent need to accordingly update waste treatment technology. The aim of this study was to determine whether a separate treatment of the liquid and the solid fraction of biowaste could enhance overall efficiency. Liquid fractions obtained from two different separation procedures were fermented at a pH of 5 and uncontrolled pH conditions for 72 h. The fermentation conditions leading to highest lactic acid productivity and yield were evaluated. The substrates gained by both separation procedures showed efficient lactic acid production up to <25 g L(-1). The pH control increased lactic acid concentration by about 27 %. Furthermore, quantitative real-time PCR analyses revealed stronger Lactobacilli growth in these fermentations. As identified via Illumina sequencing Lactobacillus delbrueckii and its closest relatives seemed to drive the fermentation independent of the substrate. These results could help to improve today's resourcing concept by providing a separate treatment of the liquid and solid biowaste fraction.

The present investigation examined the effects of supplementation of milk peptide fractions produced by enzymatic hydrolysis on the fermentation of reconstituted skim milk (RSM). Changes in pH, cell growth, proteolytic activity, and angiotensin-converting enzyme (ACE)-inhibitory activity were monitored during fermentation of RSM by pure cultures of Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, and Streptococcus thermophilus. The study showed that supplementation with peptide fractions of different molecular weights did not significantly affect the bacterial growth in RSM. All bacteria showed an increased proteolytic activity in RSM supplemented with large peptides (>10 kDa), and L. helveticus in general exhibited the highest proteolytic activity among the bacteria studied. The ACE-inhibitory activity was observed to be the maximum in RSM supplemented with larger peptides (>10 kDa) for all bacteria. The results suggest that proteolysis by bacteria leads to increased production of ACE-inhibitory peptides compared to the supplemented peptides produced by enzymatic hydrolysis.

The gut microbiota (GM) consists of resident commensals and transient microbes conveyed by the diet but little is known about the role of the latter on GM homeostasis. Here we show, by a conjunction of quantitative metagenomics, in silico genome reconstruction and metabolic modeling, that consumption of a fermented milk product containing dairy starters and Bifidobacterium animalis potentiates colonic short chain fatty acids production and decreases abundance of a pathobiont Bilophila wadsworthia compared to a milk product in subjects with irritable bowel syndrome (IBS, n = 28). The GM changes parallel improvement of IBS state, suggesting a role of the fermented milk bacteria in gut homeostasis. Our data challenge the view that microbes ingested with food have little impact on the human GM functioning and rather provide support for beneficial health effects.

Proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus is one of the key factors that determine the fermentation process and final quality of yoghurt. In this study, the interaction between different proteolytic strains of S. thermophilus and L. delbrueckii subsp. bulgaricus was investigated in terms of microbial growth, acidification and changes in the biochemical composition of milk during set-yoghurt fermentation. A complementary metabolomics approach was applied for global characterization of volatile and non-volatile polar metabolite profiles of yoghurt associated with proteolytic activity of the individual strains in the starter cultures. The results demonstrated that only non-proteolytic S. thermophilus (Prt-) strain performed proto-cooperation with L. delbrueckii subsp. bulgaricus. The proto-cooperation resulted in significant higher populations of the two species, faster milk acidification, significant abundance of aroma volatiles and non-volatile metabolites desirable for a good organoleptic quality of yoghurt. Headspace SPME-GC/MS and (1)H NMR resulted in the identification of 35 volatiles and 43 non-volatile polar metabolites, respectively. Furthermore, multivariate statistical analysis allows discriminating set-yoghurts fermented by different types of starter cultures according to their metabolite profiles. Our finding underlines that selection of suitable strain combinations in yoghurt starters is important for achieving the best technological performance regarding the quality of product., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: A lot of interesting research has been undertaken to enhance the yield of exopolysaccharides (EPS) produced by lactic acid bacteria (LAB). The objective of this study was to determine the influence of casein hydrolysates (CH) with molecular weight less than 3 kDa on cell viability, EPS synthesis and the enzyme activity involved in EPS synthesis during the co-culturing of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus in MRS broth for 72 h at 37 ± 0.1 °C., Results: The highest EPS yield (150.1 mg L⁻¹) was obtained on CH prepared with papain (CHP) at 48 h. At 24 h, EPS were composed of galactose, glucose and rhamnose in a molar ratio of 1.0:2.4:1.5. The monosaccharide composition changed with extension of the fermentation time. The activities of α-phosphoglucomutase, uridine 5'-diphosphate (UDP)-glucose pyrophosphorylase and UDP-galactose 4-epimerase were associated with EPS synthesis. Moreover, the activities of β-phosphoglucomutase and deoxythymadine 5'-diphosphate (dTDP)-glucose pyrophosphorylase involved in rhamnose synthesis were very low at the exponential growth phase and could not be detected during other given periods., Conclusion: The influence of different CH (<3 kDa) on LAB viability, EPS production, EPS monomeric composition and activity levels of key metabolic enzymes was distinct. Besides, their influence was related to the distribution of amino acids., (© 2013 Society of Chemical Industry.)

Poly-lactic acid (PLA) derived from renewable resources is considered to be a good substitute for petroleum-based plastics. The number of poly L-lactic acid applications is increased by the introduction of a stereocomplex PLA, which consists of both poly-L and D-lactic acid and has a higher melting temperature. To date, several studies have explored the production of L-lactic acid, but information on biosynthesis of D-lactic acid is limited. Pulp and corn stover are abundant, renewable lignocellulosic materials that can be hydrolyzed to sugars and used in biosynthesis of D-lactic acid. In our study, saccharification of pulp and corn stover was done by cellulase CTec2 and sugars generated from hydrolysis were converted to D-lactic acid by a homofermentative strain, L. delbrueckii, through a sequential hydrolysis and fermentation process (SHF) and a simultaneous saccharification and fermentation process (SSF). 36.3 g L(-1) of D-lactic acid with 99.8 % optical purity was obtained in the batch fermentation of pulp and attained highest yield and productivity of 0.83 g g(-1) and 1.01 g L(-1) h(-1), respectively. Luedeking-Piret model described the mixed growth-associated production of D-lactic acid with a maximum specific growth rate 0.2 h(-1) and product formation rate 0.026 h(-1), obtained for this strain. The efficient synthesis of D-lactic acid having high optical purity and melting point will lead to unique stereocomplex PLA with innovative applications in polymer industry.

Human milk oligosaccharides (HMOS) are not digested in the proximal intestine. In distal intestine, HMOS collectively modify the microbiota, but the response of individual bacteria to individual components of the HMOS is not well defined. Here, each of 25 major isolates of the human intestinal microbiota was fed individual major fucosylated and sialylated HMOS in anaerobic culture. This allowed for an assessment of the influence of specific HMOS on the growth and metabolic products of individual microbiota bacteria. Most Bifidobacteria spp. and Bacteroides spp. grew, induced α-L-fucosidase activity, and produced abundant lactate or short-chain fatty acids (SCFAs) when fed 2'-fucosyllactose (2'-FL), 3-FL, and lactodifucotetraose (LDFT). Lactobacillus delbrueckii ATCC7830, Enterococcus faecalis ATCC19433, and Streptococcus thermophilus ATCC19258 exhibited slight growth, pH reduction, and lactate production when supplemented with 2'-FL or 3-FL, but not LDFT. Supplementation with 3'-sialyllactose (3'-SL) and 6'-SL promoted moderate growth of Bifidobacterium longum JCM7007, 7009, 7010, 7011, 1272, 11347, ATCC15708, Bacteroides vulgatus ATCC8482, and B. thetaiotaomicron ATCC29148; accordingly, these bacteria exhibited greater neuraminidase activity and produced copious lactate, SCFA, or both. Lactobacillus delbrueckii ATCC7830 also consumed 6'-SL. In contrast, Clostridium spp., L. rhamnosus ATCC53103, E. faecalis ATCC29200, Staphylococcus spp., Enterobacter spp., and Escherichia coli K12 did not consume milk oligosaccharides nor produce appreciable acidic fermentation products. Specific Bifidobacteria and Bacteroides differentially digest specific individual HMOS, with the major fucosylated milk oligosaccharides most strongly stimulating key species of mutualist symbionts. This suggests strategies for treating dysbiosis of the microbiota and associated inflammatory disorders.

Mixed populations of Saccharomyces cerevisiae yeasts and lactic acid bacteria occur in many dairy, food, and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus, two microorganisms that co-occur in kefir fermentations, were studied during anaerobic growth on lactose. By combining physiological and transcriptome analysis of the two strains in the cocultures, five mechanisms of interaction were identified. (i) Lb. delbrueckii subsp. bulgaricus hydrolyzes lactose, which cannot be metabolized by S. cerevisiae, to galactose and glucose. Subsequently, galactose, which cannot be metabolized by Lb. delbrueckii subsp. bulgaricus, is excreted and provides a carbon source for yeast. (ii) In pure cultures, Lb. delbrueckii subsp. bulgaricus grows only in the presence of increased CO2 concentrations. In anaerobic mixed cultures, the yeast provides this CO2 via alcoholic fermentation. (iii) Analysis of amino acid consumption from the defined medium indicated that S. cerevisiae supplied alanine to the bacterium. (iv) A mild but significant low-iron response in the yeast transcriptome, identified by DNA microarray analysis, was consistent with the chelation of iron by the lactate produced by Lb. delbrueckii subsp. bulgaricus. (v) Transcriptome analysis of Lb. delbrueckii subsp. bulgaricus in mixed cultures showed an overrepresentation of transcripts involved in lipid metabolism, suggesting either a competition of the two microorganisms for fatty acids or a response to the ethanol produced by S. cerevisiae. This study demonstrates that chemostat-based transcriptome analysis is a powerful tool to investigate microbial interactions in mixed populations.

The mechanisms of cellular damage that lactic acid bacteria incur during freeze-thaw processes have not been elucidated to date. Fourier transform infrared spectroscopy was used to investigate in situ the lipid phase transition behavior of the membrane of Lactobacillus delbrueckii ssp. bulgaricus CFL1 cells during the freeze-thaw process. Our objective was to relate the lipid membrane behavior to membrane integrity losses during freezing and to cell-freezing resistance. Cells were produced by using 2 different culture media: de Man, Rogosa, and Sharpe (MRS) broth (complex medium) or mild whey-based medium (minimal medium commonly used in the dairy industry), to obtain different membrane lipid compositions corresponding to different recovery rates of cell viability and functionality after freezing. The lipid membrane behavior studied by Fourier transform infrared spectroscopy was found to be different according to the cell lipid composition and cryotolerance. Freeze-resistant cells, exhibiting a higher content of unsaturated and cyclic fatty acids, presented a lower lipid phase transition temperature (Ts) during freezing (Ts=-8°C), occurring within the same temperature range as the ice nucleation, than freeze-sensitive cells (Ts=+22°C). A subzero value of lipid phase transition allowed the maintenance of the cell membrane in a relatively fluid state during freezing, thus facilitating water flux from the cell and the concomitant volume reduction following ice formation in the extracellular medium. In addition, the lipid phase transition of freeze-resistant cells occurred within a short temperature range, which could be ascribed to a reduced number of fatty acids, representing more than 80% of the total. This short lipid phase transition could be associated with a limited phenomenon of lateral phase separation and membrane permeabilization. This work highlights that membrane phase transitions occurring during freeze-thawing play a fundamental role in the cryotolerance of Lb. delbrueckii ssp. bulgaricus CFL1 cells., (Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Pulsed electric field (PEF) processing involves the application of pulses of voltage for less than 1 s to fluid products placed between 2 electrodes. The effect of mild PEF on beneficial characteristics of probiotic bacteria Lactobacillus acidophilus and Lactobacillus delbrueckii ssp. bulgaricus is not clearly understood. The objective of this study was to determine the influence of mild PEF conditions on acid tolerance, growth, and protease activity of Lb. acidophilus LA-K and Lactobacillus delbrueckii ssp. bulgaricus LB-12. A pilot plant PEF system (OSU-4M; The Ohio State University, Columbus) was used. The PEF treatments were positive square unipolar pulse width of 3 µs, pulse period of 0.5s, electric field strength of 1 kV/cm, delay time of 20 µs, flow rate of 60 mL/min, and 40.5°C PEF treatment temperature. Both Lb. acidophilus LA-K and Lb. bulgaricus LB-12 subjected to mild PEF conditions were acid tolerant until the end of the 120 min of incubation, unlike the Lb. bulgaricus control, which was not acid tolerant after 30 min. The mild PEF-treated Lb. acidophilus LA-K and Lb. bulgaricus LB-12 reached the logarithmic phase of growth an hour earlier than the control. Mild PEF conditions studied significantly improved acid tolerance, exponential growth, and protease activity of both Lb. acidophilus LA-K and Lb. bulgaricus LB-12 compared with the control. The mild PEF conditions studied can be recommended for pretreating cultures to enhance these desirable attributes., (Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Three probiotic Lactobacillus strains, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus delbrueckii, were tested for their ability to assimilate and metabolize glycerol. Biodiesel-derived glycerol was used as the main carbon and energy source in batch microaerobic growth. Here, we show that the tested strains were able to assimilate glycerol, consuming between 38 and 48 % in approximately 24 h. L. acidophilus and L. delbrueckii showed a similar growth, higher than L. plantarum. The highest biomass reached was 2.11 g L⁻¹ for L. acidophilus, with a cell mass yield (Y (X/S)) of 0.37 g g⁻¹. L. delbrueckii and L. plantarum reached a biomass of 2.06 and 1.36 g L⁻¹. All strains catabolize glycerol mainly through glycerol kinase (EC 2.7.1.30). For these lactobacillus species, kinetic parameters for glycerol kinase showed Michaelis-Menten constant (K(m)) ranging from 1.2 to 3.8 mM. The specific activities for glycerol kinase in these strains were in the range of 0.18 to 0.58 U mg protein⁻¹, with L. acidophilus ATCC 4356 showing the maximum specific activity after 24 h of cultivation. Glycerol dehydrogenase activity was also detected in all strains studied but only for the reduction of glyceraldehyde with NADPH (K(m) for DL-glyceraldehyde ranging from 12.8 to 32.3 mM). This enzyme shows a very low oxidative activity with glycerol and NADP+ and, most likely, under physiological conditions, the oxidative reaction does not occur, supporting the assumption that the main metabolic flux concerning glycerol metabolism is through the glycerol kinase pathway.

Unlabelled: Continued acid production by Lactobacillus delbrueckii subsp. bulgaricus during the chilled storage of yogurt is the major cause of postacidification, resulting in a short shelf life. Two H(+) -ATPase defective variants of L. delbrueckii subsp. bulgaricus were successfully isolated and their H(+) -ATPase activities were reduced by 51.3% and 34.3%, respectively. It was shown that growth and acid production of variants were remarkably inhibited. The variants were more sensitive to acidic condition and had a significant rate for inactivation of H(+) -ATPase by N, N-dicyclohexylcarbodiimide (DCCD), along with a low H(+) -extrusion, suggesting that H(+) -ATPase is direct response for H(+) -extrusion. In addition, the variants were also more sensitive to NaCl, while H(+) -ATPase activities of variants and parent strain were significantly enhanced by NaCl stress. Obviously, H(+) -ATPase might be involved in Na(+) transportation. Furthermore, variants were inoculated in fermented milk to ferment yogurt. There was no significant difference in flavor, whereas the postacidification of yogurt during chilled storage was remarkably inhibited. It is suggested that application of L. delbrueckii subsp. bulgaricus with reduced H(+) -ATPase activity in yogurt fermentation is one of effect, economic and simple avenues of inhibiting postacidification of yogurt during refrigerated storage, giving a longer shelf life., Practical Application: During yogurt fermentation, continued acid production by Lactobacillus delbrueckii subsp. bulgaricus during the chilled storage of yogurt leads to milk fermentation with high postacidification, resulting in a short shelf life. In this work, 2 acid-sensitive variant strains of L. delbrueckii subsp. bulgaricus were isolated. The characteristics related to H(+) -ATPase were compared and it was observed that milk fermented by the variants had lower postacidification, giving a longer shelf life. Application of L. delbrueckii subsp. bulgaricus with reduced H(+) -ATPase activity in yogurt fermentation might be one of effect, economic and simple avenues of inhibiting yogurt postacidification during chilled storage, giving a longer shelf life., (© 2013 Institute of Food Technologists®)

This study examined the influence of galactooligosaccharides (GOS) and modified waxy maize starch (MWMS) addition on the growth of starter cultures, and syneresis and firmness of low-fat yogurt during storage for 28 d at 4 °C. The control yogurt (CY) was prepared without any prebiotics. Incorporation of 2.0% (w/v) GOS improved the growth of L. delbrueckii ssp. bulgaricus ATCC 11842 resulting in a shorter fermentation time. There was a significant (P < 0.05) increase in proteolysis in yogurt made with GOS (GOSY) as measured by absorbance value (0.728). Addition of GOS resulted in higher (P < 0.05) concentration of lactic and acetic acids in comparison with that of MWMSY and the CY up to day 14, thereafter, the product showed a decrease in lactic acid content in all 3 batches until the end of storage. The level of syneresis was the lowest (2.14%) in MWMSY as compared with that of GOSY (2.35%) and CY (2.53%). There was no statistically significant (P > 0.05) difference in the firmness among the 3 types of yogurt., (© 2012 Institute of Food Technologists®)

Lactobacillus delbrueckii subsp. bulgaricus 2038 (L. bulgaricus 2038) is a bacterium that is used as a starter for dairy products by Meiji Co., Ltd of Japan. Culturing L. bulgaricus 2038 with whey as the sole nitrogen source results in a shorter lag phase than other milk proteins under the same conditions (carbon source, minerals, and vitamins). Microarray results of gene expression revealed characteristics of amino acid anabolism with whey as the nitrogen source and established a model of proteolysis and amino acid biosynthesis for L. bulgaricus. Whey peptides and free amino acids are readily metabolized, enabling rapid entry into the logarithmic growth phase. The oligopeptide transport system is the primary pathway for obtaining amino acids. Amino acid biosynthesis maintains the balance between amino acids required for cell growth and the amount obtained from environment. The interconversion of amino acids is also important for L. bulgaricus 2038 growth.