Evaluation of proteolytic and ACE-inhibitory activity of Lactobacillus acidophilus in soy whey growth medium via response surface methodology

Soy whey is a rich by-product of tofu-manufacturing industries. We have previously optimized the growth of Lactobacillus acidophilus FTCC 0291 in soy whey upon supplementation of meat extract, vegetable extract and peptone using response surface methodology (RSM). The present study evaluated the proteolytic and angiotensin-I converting enzyme (ACE)-inhibitory activities of L. acidophilus FTCC 0291 in the optimized soy whey medium. The probiotic-fermented soy whey exhibited growth-associated proteolysis and ACE-inhibitory activity. Proteolysis was highly correlated with ACE-inhibitory activity, indicating that peptides liberated via fermentation may have exerted in vitro antihypertensive properties. Of the three nitrogen sources studied, peptone was found to have the highest influence on growth performance and ACE-inhibitory activity. Our results strongly indicated that probiotic-fermented soy whey produced in vitro antihypertensive bioactivity, and hence could be further developed into a carrier for probiotics with enhanced functional properties.     

Effect of prebiotics on viability and growth characteristics of probiotics in soymilk

BACKGROUND: Soy products have attracted much attention lately as carriers for probiotics. This study was aimed at enhancing the growth of probiotics in soymilk via supplementation with prebiotics. RESULTS: Lactobacillus sp. FTDC 2113, Lactobacillus acidophilus FTDC 8033, Lactobacillus acidophilus ATCC 4356, Lactobacillus casei ATCC 393, Bifidobacterium FTDC 8943 and Bifidobacterium longum FTDC 8643 were evaluated for their viability and growth characteristics in prebiotic‐supplemented soymilk. In the presence of fructooligosaccharides (FOS), inulin, mannitol, maltodextrin and pectin, all strains showed viability exceeding 7 log₁₀ colony‐forming units mL^?1 after 24 h. Their growth was significantly (P < 0.05) increased on supplementation with maltodextrin, pectin, mannitol and FOS. Additionally, supplementation with FOS, mannitol and maltodextrin increased (P < 0.05) the production of lactic acid. Supplementation with FOS and maltodextrin also increased the α‐galactosidase activity of probiotics, leading to enhanced hydrolysis and utilisation of soy oligosaccharides. Finally, prebiotic supplementation enhanced the utilisation of simpler sugars such as fructose and glucose in soymilk. CONCLUSION: Supplementation with prebiotics enhances the potential of soymilk as a carrier for probiotics. Copyright © 2009 Society of Chemical Industry     

Effect of Synbiotic Interaction of Fructooligosaccharide and Probiotics on the Acidification Profile, Textural and Rheological Characteristics of Fermented Soy Milk

Preparation of yoghurt-like product from non-dairy raw material, such as soy with probiotic and prebiotic is a novel development in the field of fermented functional foods. This research work aimed at finding the new combinations of probiotics Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus, which can give good product characteristics to fermented soy milk. Fructooligosaccharide was added in an attempt to reduce the after-taste of soymilk, improve acidification rates and growth of probiotics. Acidification rate was enhanced with L. acidophilus–L. plantarum and L. acidophilus–L. plantarum–L. rhamnosus, resulting in a shorter time to reach pH 4.5. Hardness was significantly (P?<?0.05) higher for soy yoghurt fermented by binary co-culture followed by mixed cultures. All the samples showed higher G′ (1,279.70–1405 Pa) and lower tan δ (0.273–0.346) values which signifies firmer and solid-like character of the gel formed by probiotic bacteria. Soy yoghurt made with L. acidophilus–L. plantarum resulted in improved product characteristics with shorter t _pH 4.5 (4.28 h). Soy yoghurt fermented with L. acidophilus–L. plantarum showed more than 9 log cfu/ml count which is required for probiotic functional food.     

Growth characteristics of agrowaste‐immobilised lactobacilli in soymilk during refrigerated storage

We aimed at evaluating agricultural wastes as solid supports for the immobilisation of lactobacilli in a liquid media, namely soymilk. Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733 and L. bulgaricus FTCC 0411 were immobilised on solid supports produced from durian (Durio zibethinus), cempedak (Artocarpus champeden) and mangosteen (Garcinia mangostana). The immobilised cells were inoculated into soymilk and stored at 4 °C over 168 h. Soymilk inoculated with non‐immobilised cells was used as the control. Immobilised cells showed higher survivability over 168 h compared to the control, accompanied by higher reduction of simple sugars and oligosaccharides in soymilk. Higher growth and higher utilisation of substrates also led to the higher production of lactic and acetic acids, which lowered the pH in soymilk compared to the control. Our results illustrated that agrowastes could be used as immobilisers to enhance the growth of lactobacilli in a liquid medium.     

Enhanced Growth of Lactobacilli in Soymilk upon Immobilization on Agrowastes

ABSTRACT: Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. The objective of this study was to evaluate the effects of agrowastes from durian (Durio zibethinus), cempedak (Artocarpus champeden), and mangosteen (Garcinia mangostana) as immobilizers for lactobacilli grown in soymilk. Rinds from the agrowastes were separated from the skin, dried, and ground (150 μm) to form powders and used as immobilizers. Scanning electron microscopy revealed that lactobacilli cells were attached and bound to the surface of the immobilizers. Immobilized cells of Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and L. bulgaricus FTCC 0411 were inoculated into soymilk, stored at room temperature (25 °C) and growth properties were evaluated over 168 h. Soymilk inoculated with nonimmobilized cells was used as the control. Utilization of substrates, concentrations of lactic and acetic acids, and changes in pH were evaluated in soymilk over 186 h. Immobilized lactobacilli showed significantly better growth (P < 0.05) compared to the control, accompanied by higher production of lactic and acetic acids in soymilk. Soymilk containing immobilized cells showed greater reduction of soy sugars such as stachyose, raffinose, sucrose, fructose, and glucose compared to the control (P < 0.05).     

Probiotic Strains as Starter Cultures Improve Angiotensin-converting Enzyme Inhibitory Activity in Soy Yogurt

Suitability of soy yogurt as a system for delivering probiotics and other bioactive compounds was assessed by fermenting soy milk using starter culture containing Lactobacillus delbrueckii ssp. bulgaricus Lb1466, Streptococcus thermophilus St1342, and probiotic organisms (Lactobacillus acidophilus LAFTI® L10, Bifidobacterium lactis LAFTI® B94, and Lactobacillus paracasei LAFTI® L26). Fermentations were terminated at different pH of 4.50, 4.55, and 4.60 and metabolic patterns of cultures (viability, proteolytic activity, organic acids production, angiotensin‐converting enzyme (ACE) inhibitory activity) were investigated during 28 d of storage at 4 °C. The presence of probiotics enhanced the growth of L. delbrueckii ssp. bulgaricus Lb1466 and S. thermophilus St134 in soy yogurt in comparison to the control produced by sole yogurt culture. In general, different termination pH had no effect (P > 0.05) on the viability of probiotic organisms that maintained good viability in soy yogurt during cold storage. Higher levels of essential growth factors in the form of peptides and amino acids in soy yogurts may have promoted the growth of L. acidophilus LAFTI® L10, B. lactis LAFTI® B94, and L. paracasei LAFTI® L26. The use of probiotic strains as a part of starter culture in soy yogurt resulted in a substantial increase in in vitro ACE inhibitory activity compared with the control produced by yogurt culture only. This improvement of ACE inhibition in soy yogurt is partly due to higher proteolytic activity of probiotics.     

Survival of freeze-dried microcapsules of α-galactosidase producing probiotics in a soy bar matrix

Soy oligosaccharides, mainly α-galactosides, are prevalently present in soy protein products, and can result in unfavorable digestive effects when consumed. The aim of this research was to investigate the efficiency of α-galactoside reduction by probiotic bacterial hydrolysis and if such bacteria could be maintained in a high number in a soy protein product in a microencapsulated and freeze-dried form. The probiotic bacterium, Lactobacillus acidophilus LA-2, when induced by raffinose, exhibited a high level of α-galactosidase activity at 5.0 U/mg. To preserve probiotics with high viability, cells were microencapsulated and freeze-dried. Optimization of microencapsulation presented that a combination of κ-carrageenan and inulin at a proportion of 1.9:0.1 (w:w) as capsule wall materials, significantly retained the viability of the probiotics through freeze-drying (P ≤ 0.05). Scanning electron microscopic images confirmed that the morphology of the microcapsules was well preserved after freeze-drying. Upon incorporation into soy protein bars, the freeze-dried microcapsules of L. acidophilus LA-2 remained in high numbers throughout 14 weeks of storage at 4 °C. Results of this work with the support of other studies on microencapsulation benefits indicate a promising use of freeze-dried α-galactosidase positive microencapsulated probiotics in a soy food.     

Dried Tomato‐Flavored Probiotic Cream Cheese with Lactobacillus paracasei

Abstract: A dried tomato‐flavored probiotic cream cheese (P) containing Lactobacillus paracasei Lpc‐37 was developed for the purpose of this study. The same product, but without probiotic addition (C) was used as control. Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris were used as lactic starter cultures. Chemical composition analyses and sensory tests were performed on days 1 and 7, respectively. Titratable acidity, pH value and L. paracasei population were determined every 7 d during the refrigerated storage (21 d) of the cream cheeses. The experiment and analyses were performed in triplicate, using standard methods. Probiotic population remained greater than 10⁷ CFU/g throughout the storage period, thereby characterizing the product as potentially probiotic. Cream cheeses C and P did not differ on the sensory tests, both obtaining good overall acceptance by the consumers, of which 82.6% stated that they certainly or probably would buy the product. Practical Application: Lactobacillus paracasei Lpc‐37 is a probiotic bacterium and clinical studies have shown that this microorganism beneficially affects its host. In general, dried tomato‐flavored products and cream cheese are products with good acceptance by the consumers. Thus, regular consumption of the probiotic cream cheese developed in this study may have positive effects on health and well being of people if incorporated into their diet.     

A traditional cheese from Greece to Turkey: Armola

In the present study, red capia pepper, broccoli, pumpkin, carrot purees and a probiotic culture containing Lactobacillus acidophilus were added to the conventionally produced Armola cheese to improve the functional properties of the cheese. The analyses showed that there were significant differences between the physicochemical and functional properties of the cheese samples. The samples containing broccoli had the highest antioxidant activity. Except for the control sample, L. acidophilus counts in Armola cheese samples were found to be 10⁶ log cfu/g on day 30 and the samples were found to maintain their probiotic properties until the end of the storage period.     

Probiotic lactobacilli in a semi-soft cheese survive in the simulated human gastrointestinal tract

In this study, probiotics Lactobacillus acidophilus NCFM and Lactobacillus rhamnosus HN001 in cheese were studied using models simulating the human gastrointestinal tract with the aim of investigating whether the cheese matrix affected the survival and metabolic properties of these probiotic strains. Probiotics in cheese survived in the simulated upper gastrointestinal tract model, and numbers of L. acidophilus, L. rhamnosus and total lactobacilli were increased in the colonic fermentation simulations of the probiotic cheese when compared with the non-probiotic cheese used as a control. The cheese matrix also beneficially affected cyclooxygenase-gene expression of colonocytes in a cell culture model. Freeze-dried probiotics, which were also analysed in the colonic simulator, showed similar changes in Lactobacillus numbers, although gave a stronger increase and also affected other microbial groups. These results indicate that the probiotic microbes in cheese survive in the gastrointestinal tract and that the cheese matrix does not seem to affect the probiotic survival.     

Inulin and probiotic concentration effects on fatty and linoleic conjugated acids in cream cheeses

The aim of this study was to evaluate the changes in the lipid profile, especially in conjugated linoleic acid (CLA), in 12 cream cheese batches with different prebiotic (inulin) and probiotic (Bifidobacterium animalis and Lactobacillus acidophilus) concentrations stored for 45 days at 8 ± 0.5 °C and analyzed every 15 days. A Central Rotational Composite Design was used to establish the prebiotic and probiotic concentrations. The effect of the pre- and probiotic concentrations on CLA contents (mg g⁻¹ of fat) was verified by plotting the response surface. The linear, quadratic, and interaction coefficients of the multiple linear regression for both prebiotic and probiotics were not significant (p > 0.05). No significant changes were observed neither in the lipid profile nor in the CLA content among the different batches, in which fatty acid composition and CLA concentrations were similar to the milk used in the preparation of the cream cheese and also to the control batch.     

Sensory evaluation of probiotic Minas fresh cheese with Lactobacillus acidophilus added solely or in co-culture with a thermophilic starter culture

Sensory acceptance of formulations of probiotic Minas fresh cheese was investigated. Cheeses were prepared and supplemented with Lactobacillus acidophilus (T1 – probiotic), Lactobacillus acidophilus + Streptococcus thermophilus (T2 – probiotic + starter) or produced with no addition of cultures (T3 – control). Sensory acceptance tests were performed after 7 and 14 days of storage at 5 °C, using a 9‐point hedonic scale (1 = dislike extremely; 9 = like extremely). After 7 days, no significant difference was detected among cheeses T1, T2 and T3 (P > 0.05). After 14 days, cheeses T1 and T2 presented higher acceptance and differed significantly from cheeses T3. Cheeses T3 presented significant difference between 7 and 14 days of storage (P < 0.05), whereas probiotic cheeses T1 and T2 were stable in the same period (P > 0.05). The addition of L. acidophilus, either solely or in co‐culture with a thermophilic starter culture, resulted in good acceptance of Minas fresh cheese, improving sensory performance of the product during storage.     

Optimisation of probiotic yoghurt production containing Zedo gum

A Box‐Behnken design was applied to optimise the viability of Lactobacillus acidophilus and Bifidobacterium bifidum in probiotic yoghurt containing a novel exudative Zedo gum. The effect of incubation temperature, probiotic inoculation rate, storage time and Zedo gum concentration on quality indices of the yoghurt were explored. With respect to probiotics viability, probiotic inoculation rate was the most important factor followed by the storage time. Zedo gum did not show any significant effect on probiotics viability. The optimum conditions of probiotic yoghurt production were as follows: probiotic inoculation level, 12.8 g/100 kg of milk; incubation temperature, 41.6 °C; and Zedo gum concentration, 0.13%.     

Shelf life and quality of probiotic yogurt produced with Lactobacillus acidophilus and Gobdin

This study evaluated the effect of dry white mulberry and walnut paste (Gobdin, a traditional Turkish food) in probiotic yogurt on the survival of Lactobacillus acidophilus and yogurt properties. Six different yogurts were produced with 0%, 5% and 10% Gobdin using Lactobacillus bulgaricus + Streptococcus thermophilus and with 0%, 5% and 10% Gobdin using L. bulgaricus + S. thermophilus + L. acidophilus. The physical, chemical, microbiological and sensorial properties of the yogurts were evaluated based on storage at 4 ± 1 °C. Probiotic shelf life and the most suitable combinations were determined. The highest L. acidophilus count (8.65 log cfu g^?1) was found in the 5% Gobdin‐supplemented yogurt on the 7th day of storage, while the lowest count (8.11 log cfu g^?1) was found in the probiotic control yogurt on the 21st day. Although the L. acidophilus counts in the probiotic yogurts declined during storage, all values found throughout the 21‐day storage period were >8 log cfu g^?1. This is above the level necessary to provide the desired therapeutic effect in probiotic products (10⁶–10⁷ cfu g^?1). The highest overall acceptability score was obtained on the first day from the yogurt with 5% Gobdin. However, all yogurt samples had general acceptability scores between 7 and 8 points from a 9‐point maximum. Thus, this study determined that a new functional yogurt can be produced using L. acidophilus with 5% Gobdin.     

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.     

Influence of probiotic bacteria on the proteolysis profile of a semi-hard cheese

Two probiotic strains, Lactobacillus acidophilus and Lactobacillus paracasei subsp. paracasei, were used as adjunct cultures in semi-hard cheesemaking experiments, in order to study their influence on proteolysis during ripening. Cheeses with and without probiotic bacteria were manufactured. The population of probiotics remained above 10⁷ cfu g⁻¹ during all ripening, and they did not influence primary proteolysis. However, L. acidophilus produced a significant increase in the level of low molecular weight nitrogen compounds and individual free amino acids; the amino acid profiles were also different. Multivariate analysis of peptide profiles showed that samples were grouped mainly by ripening time, although the impact of probiotics was also noticeable. L. acidophilus showed a clear influence on secondary proteolysis, while a minor effect of L. paracasei was evidenced at the end of the ripening. These results showed that the tested strains influenced distinctly proteolysis of cheeses, probably as a consequence of their different proteolytic systems and their activity via the alimentary matrix (cheese).     

Improving the viability of Bifidobacterium bifidum BB-12 and Lactobacillus acidophilus LA-5 in white-brined cheese by microencapsulation

The viability of Bifidobacterium bifidum BB-12 and Lactobacillus acidophilus LA-5 microencapsulated by either an extrusion or an emulsion technique and used in white-brined cheese was monitored. Both microencapsulation techniques were effective in keeping the numbers of probiotic bacteria higher than the level of the therapeutic minimum (>10⁷ cfu g^?1). While the counts of probiotic bacteria decreased approximately 3 log in the control cheese in which probiotics were used as free cells, the decrease was more limited in the cheeses containing microencapsulated cells (approximately 1 log). Medium- and long-chain free fatty acid contents of the cheeses with immobilized probiotics were much higher than in the control cheese. Similarly, cheeses made with immobilized probiotics contained higher acetaldehyde and diacetyl levels than the control. Experimental cheeses containing microencapsulated probiotics were not different from the control cheese in terms of sensory properties.     

Effects of 4 Probiotic Strains in Coculture with Traditional Starters on the Flavor Profile of Yogurt

To study the influence of probiotics on the flavor profile of yogurt, 4 probiotics, including Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus casei, were cofermented with traditional starters. The changes of bacterial growth, acid contents and volatile compounds of yogurt were investigated during fermentation and refrigerated storage. The strains that exhibited a low growth rate in milk did not significantly affect the bacterial population dynamics, acidity, or organic acid content during fermentation and storage. However, high viability and enhancement of postacidification were clearly observed in the samples that contained strains with a high growth rate in milk, particularly L. casei. A total of 45 volatile compounds, detected in most samples, were identified by headspace solid‐phase micro‐extraction followed by gas chromatography‐mass spectrometry. Among these compounds, ketones and aldehydes were the most abundant. The presence of either L. rhamnosus or L. plantarum did not significantly affect the major volatile compounds, while contributions of L. casei and L. acidophilus were found in the formation of minor volatile metabolites. Electronic nose measurements exhibited a good discrimination of samples that contained different probiotics during refrigerated storage.     

Removal of cholesterol by lactobacilli via incorporation and conversion to coprostanol

Fifteen strains of Lactobacillus and Bifidobacterium were screened based on their ability to adhere to hydrocarbons via the determination of cellular hydrophobicity. Lactobacillus acidophilus ATCC 314, L. acidophilus FTCC 0291, Lactobacillus bulgaricus FTCC 0411, L. bulgaricus FTDC 1311, and L. casei ATCC 393 showed greater hydrophobicity and, thus, were selected for examination of cholesterol-removal properties. All selected strains showed changes in cellular fatty acid compositions, especially total fatty acids and saturated and unsaturated fatty acids in the presence of cholesterol compared with those grown in the absence of cholesterol. In addition, we found that cells grown in media containing cholesterol were more resistant to sonication and enzymatic lysis compared with those grown without cholesterol. We further evaluated the location of the incorporated cholesterol via the insertion of fluorescence probes into the cellular membrane. In general, enrichment of cholesterol was found in the regions of the phospholipid tails, upper phospholipids, and polar heads of the cellular membrane phospholipid bilayer. Our results also showed that lactobacilli were able to reduce cholesterol via conversion of cholesterol to coprostanol, aided by the ability of strains to produce cholesterol reductase. Our results provided experimental evidence to strengthen the hypothesis that probiotics could remove cholesterol via the incorporation of cholesterol into the cellular membrane and conversion of cholesterol to coprostanol. The strains studied may be potential health adjunct cultures in fermented dairy products with possible in vivo hypocholesterolemic effects.     

Influence of probiotic strains added to cottage cheese on generation of potentially antioxidant peptides,anti-listerial activity,and survival of probiotic microorganisms in simulated gastrointestinal conditions

Our objective was to evaluate the viability of probiotic microorganisms added to cottage cheese under simulated gastrointestinal conditions, the release of potentially-antioxidant peptides, and their antimicrobial effect on Listeria monocytogenes. Cottage cheeses were prepared in triplicate, incorporating Lactobacillus casei, Lactobacillus rhamnosus GG, the commercial mix YO-MIX™ 205, or a control without probiotic addition. The probiotic population remained at >10⁶ cfu g⁻¹ during 28 days of storage at 8 °C. Cheeses made with added probiotics showed an increased metabolic activity with higher levels of lactic and acetic acids. Higher numbers of potentially bioactive peptides were observed in cheeses added with probiotics. L. monocytogenes population was reduced by about one log cycle after 20 days of storage, in cheeses with probiotics added. Our results indicate that cottage cheese is a good vehicle for probiotic bacteria.