• Journal of Microbiology and Biotechnology
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• v.31 no.6
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• pp.840-846
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• 2021

To improve the bile salt and acid tolerance of probiotics against gastrointestinal stresses, we investigated the effects of soybean lecithin and whey protein concentrate (WPC) 80 on the bile salt tolerance of Lacticaseibacillus paracasei L9 using a single-factor methodology, which was optimized using response surface methodology (RSM). The survival rate of L. paracasei L9 treated with 0.3% (w/v) bile salt for 2.5 h, and combined with soybean lecithin or WPC 80, was lower than 1%. After optimization, the survival rate of L. paracasei L9 incubated in 0.3% bile salt for 2.5 h reached 52.5% at a ratio of 0.74% soybean lecithin and 2.54% WPC 80. Moreover, this optimized method improved the survival rate of L. paracasei L9 in low pH condition and can be applied to other lactic acid bacteria (LAB) strains. Conclusively, the combination of soybean lecithin and WPC 80 significantly improved the bile salt and acid tolerance of LAB. Our study provides a novel approach for enhancing the gastrointestinal tolerance of LAB by combining food-derived components that have different properties.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.3
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• pp.433-440
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• 1998

In order to select probiotics with a high survival rate in gut and the growth inhibition of virulent pathogens to human beings or animals, we have examined a variety of microorganisms to assess the acid, bile, and pancreatic tolerance and the growth inhibition of E. colt O8 and Salmonella choleraesuis ATCC 8391. Lactobacillus acidophilus KCTC 3155 was shown to inhibit the growth of E. coli and Salmonella dramatically within 24 h of incubation, although it was vulnerable to the exposure of bile acids. Bacillus polyfermenticus showed a good growth inhibition against E. coli, with a moderate acid and bile tolerance, while Clostridium butyricum KCTC 1786 inhibited the growth of E. coli and Salmonella slightly with a good bile tolerance. However, Saccharomyces cerevisiae KCTC 7928 and Aspergillus oryzae KCTC 6075 did not inhibit the growth of E. coli and Salmonella, suggesting that these microorganisms can be used as the sources of nutritional suppliment rather than as probiotics itself.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.513-518
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• 1995

To evaluate bile salt-tolerance of lactic acid bacteria (LAB, Lactobacillus acidophilus ATCC 4356, Lactobacillus casei IFO 3533, Streptococcus thermnophilus KCTC 2185, Lactobacillus lactis ATCC 4797, and Lactobacillus bulgaricus ATCC 11842), We investigated the survivals, acid production and $\beta $-galactosidase activity of LAB under anaerobic broth system. Cellular permeability of LAB and their cellular retention of $\beta $-galactosidase were also examined in the same system. Although the growth of LAB was slightly suppressed by 0.3% bile salt, they showed normal growth curve. Streptococcus thermophilus KCTC 2185 was significantly more resistant to bile salt than the others. The $\beta $-galactosidase activity from Streptococcus thermophilus KCTC 2185 and Lactobacillus bulgaricus ATCC 11842 and their cellular retention of $\beta $-galactosidase decreased by 0.3% bile salt. The cellular permeability of LAB in the presence of bile salt increased significantly.

• Journal of Radiation Industry
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• v.6 no.2
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• pp.177-180
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• 2012

The acid and bile tolerance changes of 5 different lactic acid bacteria (LAB; Lactobacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus debruekii) with gamma irradiation were evaluated. The abilities of LAB to survive in the acidic conditions at the stomach and the bile acidic conditions at the beginning of the small intestine are the key functions for applying LAB to probiotics. In the results, all of LAB survived more than 50% after incubation in PBS (pH 2.5) for 2 hr, which indicated more than half of LAB are possible to pass through the stomach. However, gamma irradiation decreased the acid tolerances of LAB. The bile tolerances of all bacteria except Lactobacillus acidophilus were observed to survive at a 3% oxgall concentration in MRS, and 1 kGy of gamma irradiation to LAB did not affect any bile tolerances changes. But gamma irradiated Lactobacillus casei and Lactobacillus casei (3 kGy) showed decreasing survival rate with oxgall added MAS agar. In conclusion, gamma irradiation should be applied to yogurt or fermented foods with care because LAB could be changes their properties on acid and bile tolerances.

• Food Science of Animal Resources
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• v.36 no.1
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• pp.37-43
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• 2016

Cardiovascular and cerebrovascular diseases seriously harm human health, and Bifidobacterium is the most beneficial probiotic in the gastrointestinal tract of humans. This work aimed to screen cholesterol-lowering Bifidobacterium from Guizhou Xiang Pig and evaluate its tolerance to oxygen, acid, and bile. Twenty-seven aerotolerant strains with similar colony to Bifidobacterium were isolated through incubation at 37℃ in 20% (v/v) CO₂-80% (v/v) atmospheric air by using Mupirocin lithium modified MRS agar medium, modified PTYG with added CaCO₃, and modified PTYG supplemented with X-gal. Ten strains with cholesterol-lowering rates above 20% (w/w) were used for further screening. The selected strains’ tolerance to acid and bile was then determined. A combination of colony and cell morphology, physiological, and biochemical experiments, as well as 16S rRNA gene-sequence analysis, was performed. Results suggested that BZ25 with excellent characteristics of high cholesterol-removal rate of 36.32% (w/w), as well as tolerance to acid and bile, was identified as Bifidobacterium animalis subsp. lactis. To further evaluate Bifidobacterium BZ25’s growth characteristic and tolerance to oxygen, culture experiments were performed in liquid medium and an agar plate. Findings suggested that BZ25 grew well both in environmental 20% (v/v) CO₂-80% (v/v) atmospheric air and in 100% atmospheric air because BZ25 reached an absorbance of 1.185 at 600 nm in 100% atmospheric air. Moreover, BZ25 was aerotolerant and can grow in an agar medium under the environmental condition of 100% atmospheric air. This study can lay a preliminary foundation for the potential industrial applications of BZ25.

• Food Science of Animal Resources
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• v.32 no.4
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• pp.409-413
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• 2012

This study aimed to compare the probiotic characteristics of twelve strains of Lactobacillus acidophilus including cholesterol assimilation and conjugated linoleic acid (CLA) production. Cholesterol assimilation exhibited some variation among L. acidophilus strains, which could be classified into three groups based on their assimilation levels (p<0.05). The high cholesterol assimilation group exhibited a significantly higher tolerance to 0.3 and 0.5% bile acid than the low cholesterol assimilation group (p<0.05). Cholesterol assimilation showed positive correlation with 0.5% bile tolerance, and a negative correlation with acid tolerance (p<0.01). Glycocholate deconjugation activity showed no relationship with cholesterol assimilation, whereas taurocholate deconjugation activity was shown to have negative correlation with cholesterol assimilation (p<0.05). CLA production by L. acidophilus strains exhibited a wide variation, ranging from 2.69 to 5.04 mg/g fat. CLA production of L. acidophilus GP1B was the highest among the tested strains, but there was no evidence for differences in CLA production in strain specificity. Based on these results, the cholesterol assimilation of L. acidophilus strains may not be related to deconjugation activity, but may in-fact be attributed to their bile-tolerance.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.726-731
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• 2003

The aim of this research was to identify candidate probiotic lactic bacteria among indigenous dadih lactic isolates. Dadih is an Indonesian traditional fermented milk of West Sumatra which is fermented naturally. Viability of the strain is critical in determining the capacity of lactic bacteria to induce immune stimulation as well as to colonize in the intestinal tract. Therefore, LAB are proposed to exert health promoting or probiotic effects in human, such as inhibition of pathogenic microflora, antimutagenic, and the reduction of cholesterol levels. This manuscript reports in vitro probiotic properties of indigenous dadih lactic bacteria, especially some important colonization factors in GI tract, such as lysozyme, acid and bile tolerance. Bile Salt Hydrolase (BSH) activity, spectrum of bacteriocin, and antimutagenic activity of bacterial cells were also assessed. Twenty dadih lactic isolates were screened further for their tolerance to low pH, at pH 2 and 3 as well as their bile tolerance. There were ten isolates classified as acid and bile acid tolerant, and further screened for lysozyme tolerance, BSH activity. The spectrum of bacteriocin activity of isolates was assayed using cell-free neutralized supernatants by agar spot test against variety of pathogens. Lc. lactis subsp. lactis IS-10285, IS-7386, IS-16183, IS-11857 and IS-29862, L. brevis IS-27560, IS-26958 and IS-23427, Leu.mesen.mesenteroides IS-27526, and L. casei IS-7257 each has good survival rate at low pH values and in the presence of lysozyme, and short lag time in the presence of 0.3 % oxgall. Lc. lactis subsp. lactis IS-11857 and IS-29862 each has high BHS activity, Lc. lactis subsp. lactis IS-10285 and IS-16183 each had a positive spectrum of bacteriocin activity against E. coli 3301 and Lysteria monocytogenes ATCC 19112, while L. brevis IS-26958 has high BHS activity as well as positive spectrum of bacteriocin against E. coli 3301, Lysteria monocytogenes ATCC 19112, and S. aureus IFO 3060. All of the ten dadih lactic strains performed in vitro acid and bile tolerance, indicating a possibility to reach the intestine alive, and display probiotic activities.

• Journal of Dairy Science and Biotechnology
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• v.37 no.1
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• pp.40-48
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• 2019

Bifidobacteria are a prototype probiotic, which normally inhabit the intestinal tract of humans. In the present study, four species of Bifidobacterium isolated from the feces of infants were characterized. The tolerance for acid or bile salt, autoaggregation, and antibiotic resistance of the bacteria were examined. The four species were resistant to low pH, bile salts, and up to 3% bile acid. Autoaggregation rates were as high as 90%. The bacteria were consistently resistant to gentamicin, kanamycin, streptomycin, ciprofloxacin, and nalidixic acid. Due to their tolerance to environmental factors like acid and bile salts, B. longum MG723, B. breve MG729, B. bifidum MG731, and B. animalis subsp. lactis MG741 are potentially valuable as probiotics and may be useful for industrial application.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.349-352
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• 2004

Twelve strains of Lactobacillus acidophilus isolated from feces of human or animal sources were tested for probiotic properties such as cholesterol assimilation, bile and acid tolerances, and CLA production. Although the cultures showed some variation with respect to each test, the 12 strains could be classified into 3 groups based on their ability to assimilate cholesterol. The cholesterol assimilation showed positive correlation with bile tolerance and negative correlation with acid tolerance. The cholesterol assimilation of L. acidophilus strains may not be related to the deconjugation activity, but may in fact be attributed to its bile tolerance. CLA production by lactic acid bacteria (LAB) exhibited a wide variation that ranged from 2.69 to 7.64 mg/g fat. CLA production of Bifidobacterium longum ATCC 15707 was the highest among the LAB tested, but there was no evidence for differences in CLA production between genus and species.

• Korean Journal of Microbiology
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• v.47 no.3
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• pp.231-240
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• 2011

The objective of this study was to evaluate the acid and bile tolerance, bile salt hydrolase (BSH) activity, and cholesterol assimilation ability of lactic acid bacteria isolated from mustard leaf kimchi. MLK11, MLK22, MLK27, MLK41, and MLK67 were relatively acid- and bile-tolerant strains, with more than $10^5$ CFU/ml after incubation in simulated gastric juice and intestinal fluid, while MLK53 was the most sensitive strain to acid and bile. Strains MLK22 and MLK67 deconjugated the highest level of sodium glycocholate with more than 3.5 mM of cholic acid released, while deconjugation was lowest by strains MLK13 and MLK41 which released only 1.35 mM and 1.16 mM, respectively. Specially, strains MLK22 and MLK67 showed higher deconjugation of sodium glycocholate compared to sodium taurocholate and conjugated bile mixture. Although strains MLK22 and MLK67 exhibited maximal BSH activity at the stationary phase, MLK22 had somewhat higher total BSH activity compared to MLK67 towards both sodium glycocholate and sodium taurocholate. Meanwhile, cholesterol removal varied among tested strains (p<0.05) and ranged from 5.22 to 39.16 ${\mu}g$/ml. Especially, MLK67 strain assimilated the highest level of cholesterol in media supplemented with 0.3% oxgall, cholic acid, and taurocholic acid (p<0.05). According to physiological and biological characteristics, pattern of carbohydrate fermentation, and 16S rDNA sequence, strain MLK67 that may be considered as probiotic strain due to acid and bile tolerance and cholesterol-lowering effects was identified as Pediococcus pentosaceus MLK67.