• Title/Summary/Keyword: bile resistance

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Development of Bile Salt-Resistant Leuconostoc citreum by Expression of Bile Salt Hydrolase Gene

  • Cho, Seung Kee;Lee, Soo Jin;Shin, So-Yeon;Moon, Jin Seok;Li, Ling;Joo, Wooha;Kang, Dae-Kyung;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2100-2105
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    • 2015
  • Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile salt-resistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.

Crosstalk between FXR and TGR5 controls glucagon-like peptide 1 secretion to maintain glycemic homeostasis

  • Kim, Hyeonhui;Fang, Sungsoon
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.140-146
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    • 2018
  • Though bile acids have been well known as digestive juice, recent studies have demonstrated that bile acids bind to their endogenous receptors, including Farnesoid X receptor (FXR) and G protein-coupled bile acid receptor 1 (GPBAR1; TGR5) and serve as hormone to control various biological processes, including cholesterol/bile acid metabolism, glucose/lipid metabolism, immune responses, and energy metabolism. Deficiency of those bile acid receptors has been reported to induce diverse metabolic syndromes such as obesity, hyperlipidemia, hyperglycemia, and insulin resistance. As consistent, numerous studies have reported alteration of bile acid signaling pathways in type II diabetes patients. Interestingly, bile acids have shown to activate TGR5 in intestinal L cells and enhance secretion of glucagon-like peptide 1 (GLP-1) to potentiate insulin secretion in response to glucose. Moreover, FXR has been shown to crosstalk with TGR5 to control GLP-1 secretion. Altogether, bile acid receptors, FXR and TGR5 are potent therapeutic targets for the treatment of metabolic diseases, including type II diabetes.

Probiotic Properties of Bifidobacteria Isolated from Feces of Infants (유아 분변에서 분리한 비피도박테리아의 프로바이오틱스 기능성 연구)

  • Kang, Chang-Ho;Kim, YongGyeong;Han, Seul Hwa;Jeong, Yulah;Park, HyeMin;Paek, Nam-Soo
    • 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.

Antibacterial Activity and Probiotic Properties of Lactic Acid Bacteria from Korean Intestine Origin (한국인 장관에서 분리한 유산균의 항균활성 및 프로바이오틱스 특성 연구)

  • Kang, Chang-Ho;Kim, Yong Gyeong;Han, Seul Hwa;Jeong, Yulah;Paek, Nam-Soo
    • KSBB Journal
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    • v.32 no.2
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    • pp.153-159
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    • 2017
  • The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from human feces. The properties were tested on the basis of guideline for probiotic selection protocol such as tolerance for acid or bile salt, autoaggregation, antibiotic resistance, and antimicrobial activity. Total 25 lactic acid bacteria were isolated from human feces, and their antibacterial activity was tested against Staphylococcus aureus, Escherichia coli, E. coli O157:H7, Vibrio parahaemolyticus, V. alginolyticus using an agar diffusion assay. Among them, 4 selected strains were identified by analysis of their 16S rRNA, as Lactobacillus rhamnosus MG316, L. acidophilus MG501, L. reuteri MG505, and L. gasseri MG570. Results show that resistance to low pH and bile salts. Also, the selected strains were resistant to bile acid up to 3% and their autoaggregation rates were as high as 60%. All strains tested were resistance to nalidixic acid and kanamycin.

Isolation and Identification of Lactobacillus plantarum CIB 001 with Bile Salt Deconjugation Activity from Kimchi (김치로부터 담즙산 분해능이 우수한 Lactobacillus plantarum CIB 001의 분리 및 동정)

  • Cha, Sang-Do;Kim, Tae-Woon;Lee, Dong-Hee
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.222-226
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    • 2010
  • This study was carried out to isolate and characterize the Lactobacillus plantarum with bile salt deconjugation activity that was isolated from Kimchi. Some isolates were selected and identified as L. plantarum by 16S rRNA gene sequence and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of whole cell protein patterns. They were assayed to determine their capacities to express bile salt hydrolase (BSH) activity. Among the identified strains, L. plantarum CIB 001 showed the highest level of BSH activity. Then, resistance to gastric acidity and bile condition were analyzed for further characterization. This strain was able to maintain viability for 1h at pH 2.0 and to survive in a MRS (deMan, Rogosa, and Sharpe) broth with 1.0% of bile acids. L. plantarum CIB 001 would potentially be useful in the food industry as probiotics.

Proteomic Analysis of Proteins of Weissella confusa 31 Affected by Bile Salts

  • Lee, Kang Wook;Lee, Seung-Gyu;Han, Nam Soo;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1432-1440
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    • 2012
  • Weissella confusa 31, an isolate from human feces, possesses desirable properties as a probiotic strain, including bile salt resistance. W. confusa 31 is not inhibited by bile salts up to 0.3% concentration. Proteins affected by bile salts (0.05%) were examined by 2-D gel electrophoresis. Our proteomic analyses revealed that the intensities of 29 spots were changed, where 17 increased (including 2 spots observed only under the bile salts stress conditions) and 12 decreased. Proteins were identified by MALDI-TOF mass spectrometry. Proteins increased in the band intensities included adenylate kinase (12.75-fold increase), Clp-like ATP-dependent protease (11.91-fold), 6-phosphogluconate dehydrogenase (10.35-fold), and HSP 70 (5.07-fold). Some of the increased or decreased proteins are also known to be involved in other types of stress responses.

In vitro selection of lactic acid bacteria for probiotic use in pigs (양돈용 생균제 개발을 위한 유산균주 선발)

  • Byun, Jae-won;Kim, Gyung-tae;Bae, Hyoung-suk;Baek, Voung-jin;Lee, Wan-kyu
    • Korean Journal of Veterinary Research
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    • v.40 no.4
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    • pp.701-706
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    • 2000
  • This study was carried out to select the lactic acid bacteria(Lactobacillus, Streptococcus and Bifidobacterium) and yeast for probiotic use in pigs. Acid-tolerant 536 strains were isolated from the feces of 30 pigs. To select useful strains, the first screened strains were treated with strong acid solution(pH 2.5 to 3.0) for 3 hours and subsequentely treated with the anaerobic diluent solution containing 0.15% Oxgall for 3 hours. Among these strains, 151 strains showed strong tolerance to both acid and bile. Lactobacillus and Streptococcus tolerant to the acid and bile were treated with heat at $80^{\circ}C$ for 15 min, and at $70^{\circ}C$ for 5 min in Bifidobacterium and yeast. As a result of heat treatment, 38 strains were obtained as heat-tolerant strains. All of heat-tolerant strains were tested for antibiotic resistance against virginiamycin, sulfathiazole, aureomycin, neomycin, linsmycin, tiamulin and ASP250 which were used as feed additives for growth promotion in pigs. Finally, one strain each from Lactobacillus, Streptococcus, Bifidobacterium and yeast that showed resistance to acid, bile, heat and antibiotics was selected for probiotic use in pigs.

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Some Probiotic Properties of Some Lactic Acid Bacteria and Yeasts Isolated from Jeot-gal. (젓갈 프로바이오틱 생균의 내산성 및 내담즙 특성)

  • 이나경;김현욱;최신양;백현동
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.297-300
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    • 2003
  • 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 screened lactic acid bacteria and yeasts from Jeot-gal to assess resistance against the artificial gastric acid and bile juice. Lactic acid bacteria and yeasts isolated were incubated for 24 h in artificial bile juice after incubation for 2 h in artificial gastric acid. Especially, strain HW 161 and strain NK 181 showed the higher survival for 2 h incubation in artificial gastric acid. All of 3 strains of lactic acid bacteria and 2 strains of yeast were showed higher growth rate than the control in artificial bile. The antimicrobial activity of lactic acid bacteria and yeasts was also investigated to prove efficacy as probiotic organisms. Lactic acid bacteria were shown the inhibition of Gram positive and negative bacteria, but yeasts narrow inhibition.

김치에서 분리한 유산균의 인공위액과 담즙에서의 생존특성과 항균성

  • 이신호;노명자
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.617-622
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    • 1997
  • This studies were carried out to investigate the physiological characteristics of lactic acid bacteria related to kimchi fermentation. 157 strains of lactic acid bacteria were isolated from home-made kimchi of Taegu district. 144 strains of isolated lactic bacteria showed resistance to artifical gastric juice and bile. Among isolated strains, fifteen strains were selected according to the growth characteristics. The selected strains survived for 24 hours in artifical bile after incubation for 3 hours in artificial gastric jucie (pH 2.5). Especially, three strains such as B-10, K-2, and F-3, showed little viable cell change for 3 hrs incubation in artifical gastric juice. The selected strains showed low viability in artificial gastric juice but high in artifical bile. The selected strains also showed significantly antimicrobial activities to pathogens such as Listeria monocytogenes, Escherichia coli, Bacillus subtili, Staphylococcus aureus and Vibrio parahaemolyticus. The fifteen selected strains were identified as Lactobacillus plantarum, Lactobacillus sake and Leuconostoc mesenteroides.

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Changing Patterns of Serum and Bile Antibodies in Re-infected Rats with Clonorchis sinensis

  • Zhang, Hong-Man;Chung, Byung-Suk;Li, Shun-Yu;Choi, Min-Ho;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.46 no.1
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    • pp.17-22
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    • 2008
  • Rats develop strong resistance to re-infection and super-infection by Clonorchis sinensis. The present study investigated the antibodies present in the sera and bile juice of rats that were primary infected and re-infected with C. sinensis. The serum level of specific IgG antibodies, which were elevated 2 wk of the primary infection, peaked at 4 wk and subsequently remained unchanged even during re-infection. The total IgE level in serum increased slowly from 388 ng / ml to 3,426 ng / ml beginning 2 wk after the primary infection, and remained high up to 8 wk but dropped to a normal level (259 ng / ml) after treatment. In resistant re-infected rats, the serum IgE level increased rapidly and peaked within 1 wk, whereas no increase was observed in immunosuppressed rats. The serum level of specific IgA antibodies was elevated beginning 1 wk after infection, and decreased 4 wk after treatment. The total bile IgA level unchanged during the primary infection but increased in treated and re-infected rats. The elevated levels of serum IgE and bile IgA indicate that these immunoglobulins may be correlated with the development of resistance to re-infection by C. sinensis in rats.