Survival of Lactobacillus casei and functional characteristics of reduced sugar red beetroot yoghurt with natural sugar substitutes

In this study, probiotic gel fermentation and functional characteristics of reduced sugar yoghurt with red beetroot bioactive components and steviol glycosides from Stevia rebaudiana extracts as natural source of noncaloric sweeteners were investigated. A significant increase in Lactobacillus casei viability and fermented gel network and a decrease in syneresis were determined in yoghurt sweetened with steviol glycosides and high content of red beetroot bioactive components such as total phenolics, anthocyanins and total antioxidant capacity (DPPH and FRAP). The survival of L. casei was maintained within probiotic and therapeutic levels (>9 log₁₀ CFU/g), due to the possible prebiotic effect of steviol glycosides and fermentable fibres and phenolic compounds of red beetroot during 28 days of cold storage of the probiotic yoghurts. Colour properties (L*, a*, b*, ΔE*, C*, h*) and such sensory properties as smoothness, sweetness astringency were significantly affected. The highest organic acid content was found in red beetroot enriched yoghurt with stevia, while the most dominant organic acids were butyric, lactic, malic and citric. As a result, it was determined that fermented milk products, which are supported by phytochemical and steviol glycoside interactions, have adequate nutritional effects, high probiotic viability and acceptable sensory properties     

Antimicrobial and antidiabetic potential of synbiotic fermented milk: A functional dairy product

The current study was based on evaluation of the therapeutic potential (antimicrobial and antidiabetic) of a synbiotic fermented milk product developed by optimising levels of the prebiotic ingredients (fructooligosaccharide and isomaltooligosaccharide). The maximum growth rate (0.27 ± 0.21a cfu/mL) of Lactobacillus acidophilus ATCC^® 4357? was observed with 2.345–2.445% of fructooligosaccharide and 2.53–2.62% of isomaltooligosaccharide. Significant antimicrobial potential of the synbiotic fermented milk was observed against Escherichia coli and Staphylococcus aureus due to the addition of prebiotics. Similarly, the final product also showed 62.9%, 71.5% and 57.0% reduction in blood glucose, urea and creatinine levels, respectively, of diabetic rabbits, when supplemented with 6% of the synbiotic fermented milk. A synbiotic fermented milk product with significant therapeutic potential could be developed using fructooligosaccharide and isomaltooligosaccharide as prebiotic ingredients.     

The effects of ratio of cow's milk to soymilk,probiotic strain and fruit concentrate on qualitative aspects of probiotic flavoured fermented drinks

The effects of proportion of cow's milk to soymilk (100:0, 75:25, 50:50, 25:75, 0:100), probiotic bacteria (Lactobacillus acidophilus LA‐5 or Lactobacillus casei L‐01) and natural fruit concentrates (strawberry, apricot, peach and pear) on quality characteristics of soy‐based probiotic drink were investigated. The parameters were analysed at the end of fermentation and during 21 days of storage at 5 ^°C. The highest viability was observed when the equal proportion of cow's milk and soymilk and L. casei was used (50:50‐CY). During chilled storage, the flavouring apricot had the highest stimulatory effect on the survival of L. casei in 50:50 treatment. In general, the treatment 50:50‐CY was realised as the best one overall.     

Lactobacillus casei starter culture improves vitamin content,increases acidity and decreases nitrite concentration during sauerkraut fermentation

To investigate the effects of Lactobacillus casei on Chinese sauerkraut fermentation, L. casei 11MZ‐5‐1 was inoculated into Chinese sauerkraut. Physicochemical indexes were measured, and the microbial dynamics during the fermentation were analysed by quantitative real‐time PCR (qRT‐PCR) and polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE). The result showed that inoculation with L. casei 11MZ‐5‐1 lowered the pH of the fermentation system more rapidly than in the control model (CK). The content of vitamin C (V_C) (44.64 ± 2.12 mg kg^?1) was higher and nitrite (under 0.76 mg kg^?1) was lower than CK (P < 0.05). The numbers of 16S rRNA gene copies in the experimental model (BA) were lower than in CK at the end fermentation time. According to the PCR‐DGGE analyses, 22 and 17 specific bands were detected in CK and BA, respectively. L. paracasei and L. casei were predominant during the fermentation in BA. The relative abundance and diversity indices of bacteria in BA were 8.23 ± 0.25 and 2.01 ± 0.06, respectively, lower than in the CK (P < 0.05). So, the L. casei 11MZ‐5‐1 inoculations could effectively inhibit the microbial diversity in the fermentation system. The fermented cabbage with L. casei 11MZ‐5‐1 was more favourably estimated by consumers in terms of colour, crispness, sourness, aroma, bitterness, stink, stale flavour and overall acceptability.     

Effects of nitrates and nitrites on Lactobacillus helveticus and Lactobacillus casei dairy cultures

 The aim of this work was to observe the effects of nitrites and nitrates on the titratable acidity of milk after addition of Lactobacillus helveticus (TX 121) and L. casei (CAD 154) dairy cultures and on the levels of nitrate and nitrite. After adding L. helveticus and nitrites the increasing concentrations of the latter brought about a marked decrease in titratable acidity. In milk containing 100 mg·kg^–1 nitrite the resulting concentration was 39.33 mg·kg^–1 NaNO₂. Nitrates caused a less obvious decrease in titratable acidity, giving 13.27 g·l^–1 lactic acid. In milk containing 100 mg·kg^–1 NaNO₃ the resulting concentration was 24.99 mg·kg^–1 NaNO₃. Experiments with L. casei and a nitrite additive revealed a decrease in titratable acidity to 8.89 g·l^–1 lactic acid. After incubation, nitrite levels were reduced from 100 mg·kg^–1 NaNO₂ to 37.81 mg·kg^–1 NaNO₂. Nitrates were also stated to inhibit the titratable acidity of the sample, which decreased to 11.42 g·l^–1 lactic acid. Nitrates were reduced to 46.99 mg·kg^–1 NaNO₃. The present study shows that nitrites, more than nitrates decrease the titratable acidity of milk after addition of L. helveticus and L. casei dairy cultures. A reduction of nitrates and nitrites in milk samples by Lactobacillus was also found. The results of this study can be used in the dairy industry in the production of several types of hard cheese as well as fermented milk products that use L. helveticus and L. casei dairy cultures. Received: 31 March 1999 / Revised version: 6 September 1999     

Antioxidant activity of prebiotic ginseng polysaccharides combined with potential probiotic Lactobacillus plantarum C88

The potential prebiotic properties of Panax ginseng polysaccharides were studied using ten strains of Lactobacillus plantarum isolated from traditional Chinese fermented foods. These probiotics showed different growth characteristics depending on the extract and strain specificity. L. plantarum C88 showed higher cell densities and growth rate when cultured on P. ginseng polysaccharides. In the in vitro antioxidant assay, P. ginseng polysaccharides combined with L. plantarum C88 were found to possess significant DPPH, ABTS and superoxide anion radicals scavenging activities, and acidic polysaccharides showed better antioxidant activity than neutral polysaccharides. Furthermore, we evaluated the antioxidant effect of acidic P. ginseng polysaccharide combined with L. plantarum C88 strain in natural ageing mice in vivo. Acidic P. ginseng polysaccharide and L. plantarum C88 together inhibited the formation of malondialdehyde (MDA) and increased the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px), catalase (CAT) and total antioxidant capacities (T‐AOC) in a dose‐dependent manner.     

Effect of nisin‐producing Lactococcus lactis starter cultures on the inhibition of two pathogens in ripened cheeses

The effect of Lactococcus lactis nisin‐producing strains, isolated from Italian fermented foods, on the survival of two foodborne pathogens namely Listeria monocytogenes and Staphylococcus aureus was investigated in experimental cheese production. One of the three Lactobacillus lactis nisin innoculated as starters, Lactobacillus lactis 41FL1 lowered S. aureus count by 1.73 log colony‐forming units (cfu)/g within the first 3 days, reaching the highest reduction, 3.54 log cfu/g, by the end of ripening period of 60 days. There was no effect on L. monocytogenes. The application of L. lactis 41FL1 as bioprotective culture in controlling S. aureus shows considerable promise.     

Effects of fermentation conditions on the potential anti‐hypertensive peptides released from yogurt fermented by Lactobacillus helveticus and Flavourzyme®

This study investigates the effects of fermentation conditions on the production of angiotensin‐converting enzyme inhibitory (ACE‐I) peptides in yogurt by Lactobacillus helveticus 881315 (L. helveticus) in the presence or absence of Flavourzyme^®, which is derived from a mould, Aspergillus oryzae and used for protein hydrolysis in various industrial applications. Optimal conditions for peptides with the highest ACE‐I activity were 4% (v/w) inoculum size for 8 h without Flavourzyme^® supplementation, and 1% inoculum size for 12 h when combined with Flavourzyme^®. The yogurt fermented by L. helveticus resulted in IC₅₀ values (concentration of inhibitor required to inhibit 50% of ACE activity under the assayed conditions) of 1.47 ± 0.04 and 16.91 ± 0.25 mg mL^?1 with and without Flavourzyme^® respectively. Seven fractions of ACE‐I peptides from the yogurt incorporated with L. helveticus and Flavourzyme^® were separated using the preparative high‐performance liquid chromatography. Fraction (F3) showed the highest ACE‐I activity with an IC₅₀ of 35.75 ± 5.48 μg mL^?1. This study indicates that yogurt may be a valuable source of ACE‐I peptides, which may explain the outcomes observed in the experimental and clinical studies and foresee the application of fermented milk proteins into functional foods or dietary supplements.     

Anti‐inflammatory and cytotoxic effects of ginseng extract bioconverted by Leuconostoc mesenteroides KCCM 12010P isolated from kimchi

A bioconversion technique using microorganisms has been applied to ginseng to increase content of bioactive ginsenoside and biofunctionality such as anticancer, anti‐obesity and antioxidant activities. The objective of this study was to screen lactic acid bacteria for bioconversion of ginsenosides and to evaluate anti‐inflammatory and cytotoxic effects of bioconverted ginseng extract. Strains isolated from kimchi were screened for their β‐glucosidase activities using esculin agar. Selected strain was identified based on 16S rRNA sequencing and carbohydrate fermentation. During ginseng fermentation, viable cell number and pH were determined. Bioconverted ginsenosides were analysed by HPLC. Anti‐inflammatory effects were evaluated using RAW 264.7 cells, and cytotoxic effects were determined by MTT assay. Among 166 isolates screened, Leuconostoc mesenteroides was selected for ginseng bioconversion, as it showed a higher β‐glucosidase activity and viable cell number than any of the other tested strains. After fermentation for 2 days, viable cell number was 8.8 log CFU mL^?1 and final pH was 4.8. Ginsenoside Rb₂ was bioconverted into ginsenoside Rg₃ (Rb₂ → Rd → Rg₃) by L. mesenteroides. The nitric oxide contents of 2‐day‐fermented extract decreased by as much as 25%, compared to a non‐fermented extract. The cell viabilities of HepG2, HT‐29, HeLa and LoVo treated with fermented ginseng extract also decreased by 49.7%, 20.2%, 21.0% and 8.7%, respectively, compared to those of control cells treated with non‐fermented extract. Ginseng extract bioconverted by L. mesenteroides showed anti‐inflammatory and anticancer effects. Therefore, bioconverted ginseng extract might have applications in the pharmaceutical and/or functional food industry.     

Production,mode of action and sequencing of the corresponding gene of a bacteriocin produced by Lactococcus lactis 19.3

Lactococcus lactis 19.3 produces a bacteriocin with a wide inhibitory spectrum, including the food pathogen Listeria monocytogenes. The producing strain was able to grow and produce similar amounts of bacteriocin in MRS medium, cow's milk and soya milk, respectively. The mode of action of this bacteriocin was further investigated using two indicator strains, Lactobacillus delbrueckii subsp. bulgaricus LMG 6901^T and Listeria monocytogenes ATCC 1911‐1. The bacteriocin displayed a bactericidal effect, causing a rapid decrease of the cell viability. Bacteriocin treatment of the sensitive strains resulted in major morphological changes, including pore formation, as shown by scanning electron microscopy. Finally, the bacteriocin‐encoding gene was identified by sequencing. The presence of nisin gene was confirmed. Based on all our data gathered so far, L. lactis 19.3 is a good candidate for a starter or protective culture in the manufacturing of both fermented dairy and vegetarian food products.     

Potato juice fermented with Lactobacillus casei as a probiotic functional beverage

This study was conducted to evaluate probiotic potato juice as a potential substrate for the production of Lactobacillus casei, and the change in the functionality of potato juice was monitored during fermentation. L. casei grew well in potato juice without nutrient supplementation, and lactic acid bacteria of fermented ‘Haryoung’ juice reached 1.7×10⁹ CFU/mL after a 48 h fermentation. DPPH radical scavenging activities of the potato juices decreased after a 72 h fermentation, but fermented colored potato juice still maintained >50% radical scavenging activity. The survival rate of L. casei fermented in’ Haryoung’ juice was 89.0% after exposure to an acidic condition, and L. casei in all fermented potato juice samples showed the ability (50–85%) to survive in the presence of bile. These results suggest that fermented potato juice might serve as a probiotic functional beverage for vegetarians or consumers who are allergic to dairy products.     

The biological activity of fermented milk produced by Lactobacillus casei ATCC 393 during cold storage

Lactobacillus casei ATCC 393 is an important probiotic strain widely known in dairy technology. However, its capability to produce bioactive peptides from milk proteins has not been studied. The viability of the Lb. casei ATCC 393 strain and some physicochemical properties in fermented milk throughout storage for 21 days at 4 °C was evaluated; biological activity, i.e., antioxidant, angiotensin converting enzyme inhibitory and anticancer activities of water soluble extract and its filtrate (< 2 kDa; F1) were determined. Lb. casei counts remained over 9 log cfu g⁻¹ during the storage period in fermented milk. These bioactivities were increased significantly (P < 0.01) during storage. F1 of fermented milk after three weeks of storage showed the highest bioactivity impact. De novo sequencing assay for peptide identification was applied to the mass spectrum of F1. The promising capability of Lb. casei ATCC 393 to release bioactive peptides from milk proteins was demonstrated.     

Production of propionic acid in a fermented dairy beverage

A dairy beverage containing propionic acid was produced using the adjunct starter cultures with Lactobacillus acidophilus and Propionibacterium freudenreichii ssp. shermanii. The impacts of temperature (30, 35 and 40 °C) and inoculation ratio (1:2, 1:4 and 1:8) on propionic acid production and viability of micro‐organisms were studied. The incubation temperature had a significant adverse (P < 0.05) effect on the viability of P. freudenreichii and propionic acid production. Maximum amount of produced propionic acid (in 1:4–30 °C treatment) was 0.77%w/w. P. freudenreichii and L. acidophilus counts remained high (9 × 10⁶ and 1.5 × 10⁶ cfu/mL, respectively) after cold storage.     

Proteolysis in dry‐aged loins manufactured with sonicated pork and inoculated with Lactobacillus casei ŁOCK 0900 probiotic strain

Selected parameters related to proteolysis were evaluated in dry‐aged loins manufactured with sonicated pork and inoculated with Lactobacillus casei ?OCK 0900 probiotic strain. Dehydration, pH, water activity and total nitrogen content were not affected by ultrasound treatment. Neither sonication nor inoculation has an influence on L* and a* colour parameters. Sonication significantly influenced the pattern of proteolysis providing higher nonprotein nitrogen content (NPN) and proteolysis index (PI). The highest intensity of proteolysis, as assessed by NPN and PI, occurred in an inoculated sample. Neither sonication nor inoculation with L. casei had a significant effect on total viable counts. Loins inoculated with L. casei had the highest lactic acid bacteria count, followed by the sonicated and nonsonicated control. This research shows that sonication followed by inoculation with L. casei ?OCK 0900 could be used as an effective measure to speed up the proteolytic changes in dry‐aged meat cuts.     

Effect of inulin on growth and bacteriocin production by Lactobacillus plantarum in stationary and shaken cultures

The prebiotic effect of inulin added to MRS medium on growth and bacteriocin production by L. plantarum ST16 Pa was investigated in stationary cultures in anaerobic jars with medium containing 0.025% sodium thioglycolate or in flasks shaken at 100 rpm. In the presence of 1% inulin in anaerobic stationary cultures, this strain produced lactic acid at a level that was 36.5% higher than in the absence of the polysaccharide. In shaken cultures without inulin, cell count was 54% higher than in the stationary ones. Under stationary conditions in anaerobic jars, the addition of inulin increased the maximum specific growth rate from 0.37 to 0.49 h^?1 and reduced the generation time from 1.85 h to 1.40 h. Consequently, the exponential phase was shortened from 12 to 9 h when the cells were grown in stationary cultures with the oxygen scavenger. Despite this effect of inulin on growth rate, stationary cultures without inulin displayed higher antimicrobial activity against Listeria monocytogenes L104 (3200 AU/mL) than cultures with inulin (1600 AU/mL); therefore, inulin behaved as a compound able to accelerate growth rather than to stimulate bacteriocin production. The results presented in this study are very promising, as L. monocytogenes is a well‐known foodborne pathogenic microorganism. Moreover, L. plantarum ST16 Pa has proven to be a potential producer of a natural food preservative at an industrial level.     

In vitro comparison of probiotic properties of Lactobacillus casei Zhang, a potential new probiotic, with selected probiotic strains

Since 2004, our research group has isolated 240 Lactobacillus strains from Koumiss, a traditional fermented alcoholic beverage prepared from mare's milk in Inner Mongolia, Xinjiang of China and Mongolia. Among these Lactobacillus strains a novel strain with potential probiotic properties, Lactobacillus casei Zhang, was screened out and studied extensively for its probiotic properties, health-promoting effects and fermentation characteristics. In vitro tests indicated that L. casei Zhang had high tolerance to simulated gastric, intestine juices and bile salts, similar to commercial probiotic strains such as Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus GG, L. casei Shirota and Bifidobacterium animalis Bb12. Higher acid-production activity and proteolytic activity was observed in the fermented milk inoculated with L. casei Zhang during the refrigerated storage than in the samples inoculated with the selected commercial probiotics. The yogurt samples fermented with L. casei Zhang exhibited similarer viable count (1.0 × 10⁹ cfu/mL) as the other samples after 28 d of refrigerated storage. The results suggested that L. casei Zhang showed good potential for application in functional foods and health-related products.     

Raffinose family oligosaccharides in seed of Glycine max cv. Chiang Mai60 and potential source of prebiotic substances

Raffinose family oligosaccharides (RFOs) content in Glycine max seed of cultivar Chiang Mai60, a local soybean of Thailand, was investigated. RFOs and other low molecular weight sugars were extracted by 50% (v/v) ethanol and quantified by high‐performance liquid chromatography (HPLC). The prebiotic property of this extract was subsequently studied by in vitro method. The results showed that the concentrations of raffinose, stachyose and verbascose were 6.74 ± 1.62, 145.32 ± 18.74 and 1.60 ± 0.52 mg g^?1 dry seed, respectively, while glucose and sucrose were detected at 10.73 ± 1.35 and 13.28 ± 2.16 mg g^?1 dry seed, respectively. The growth of four Lactobacilli probiotics were increased significantly in a basal liquid medium supplemented with this ethanolic extract as carbon source compared to glucose supplementation. Subsequently, defined mixed culture was studied and it was found that growth stimulation of total Lactobacilli by extracted sugars resulted in the suppression of Escherichia coli and Salmonella enterica serovar Typhimurium growth. It could be concluded that this cultivar showed the RFOs‐rich content and a potential to be a source of an effective prebiotic substance for food application.     

In vitro assessment of antimicrobial efficacy of the D-tagatose and lactobacilli-based synbiotic preparations against the pathogenic Escherichia coli and Salmonella typhimurium

The study aimed to investigate the utilisation of prebiotic D-tagatose by probiotic lactobacilli (L. rhamnosus GG, L. casei, L. acidophilus and L. fermentum) and enteric pathogens (E. coli and S. typhimurium) to determine synbiotic potential of D-tagatose and lactobacilli combination. The antimicrobial efficacy of the synbiotic preparations (D-tagatose with lactobacillus) was assessed against selected pathogens in co-culture assays. Evidently, D-tagatose supported growth of selected lactobacilli, especially the L. rhamnosus GG and L. casei, but not the enteric pathogens. The tested synbiotic preparations completely inhibited growth of both the pathogens, even in the presence of D-glucose in co-cultures. The well-diffusion assay demonstrated the presence of antimicrobial activity as recorded by drop of culture pH. The study substantiated that D-tagatose could be an effective prebiotic component for formulation of potential synbiotic combinations with L. rhamnosus GG or L. casei that can be used as an alternative to antibiotics against enteric pathogens.     

Functionality of freeze-dried L. casei cells immobilized on wheat grains

Lactobacillus casei cells were immobilized on wheat grains and the effect of nine cryoprotectants during freeze-drying was investigated. Survival and fermentative activity of the freeze-dried immobilized biocatalysts was studied by monitoring pH, lactic acid and lactose content in successive fermentations batches of both synthetic lactose medium and milk. Freeze-dried L. casei cells immobilized on wheat grains without using cryoprotectants resulted in high cell survival and metabolic activity. The same biocatalysts were stored at room temperature for 9 months and at 4 °C and −18 °C for 12 months. Reactivation of the stored biocatalysts was carried out in synthetic lactose medium. Storage at room and low temperatures (4 °C and −18 °C) resulted in about 5.11, 4.9 and 4.3 final pH respectively during fermentations, indicating the suitability of the immobilized biocatalysts for the production of mild and low pH dairy products. The immobilization of a probiotic microorganism, such as L. casei, on boiled wheat which contains prebiotic compounds might provide a potential synbiotic preparation.     

Cranberry seed fibre: a promising prebiotic fibre and its fermentation by the probiotic Bacillus coagulans MTCC 5856

Cranberry, a versatile fruit, is known for nutritional as well as medicinal properties due to the presence of bioactive compounds. However, cranberry fruit has not been fully explored for its prebiotic potential. Therefore, this study was conducted to evaluate prebiotic potential of cranberry seed fibre (CSF) and also evaluate its fermentability by the probiotic strain Bacillus coagulans MTCC 5856. The resistance to gastric acid and porcine pancreatic enzymatic (PPE) hydrolysis of CSF was investigated using an in vitro model. It was found that CSF was resistant to gastric acid and also nondigestible to PPE hydrolysis. CSF as sole nutrition source was evaluated for the fermentability by B. coagulans MTCC 5856. A significant amount of short‐chain fatty acids was produced by the B. coagulans MTCC 5856 while fermenting cranberry fibres anaerobically. CSF supported the growth of B. coagulans MTCC 5856 and also inhibited the growth of E. coli ATCC 25922 when cocultured in an anaerobic environment. CSF from the cranberry fruit exhibited prebiotic potential and also found to be fermentable by B. coagulans MTCC 5856. This study provided the scientific evidence of CSF as a prebiotic fibre and also its suitability with the probiotic B. coagulans MTCC 5856 for an ideal synbiotic preparation.