• Title/Summary/Keyword: Bile and Acid Tolerance

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Changes in Cell Membrane Fatty Acid Composition of Streptococcus thermophilus in Response to Gradually Increasing Heat Temperature

  • Min, Bonggyu;Kim, Kkotnim;Li, Vladimir;Cho, Seoae;Kim, Heebal
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.739-748
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    • 2020
  • In this study, a method of heat adaptation was implemented in an attempt to increase the upper thermal threshold of two Streptococcus thermophilus found in South Korea and identified the alterations in membrane fatty acid composition to adaptive response to heat. In order to develop heat tolerant lactic acid bacteria, heat treatment was continuously applied to bacteria by increasing temperature from 60℃ until the point that no surviving cell was detected. Our results indicated significant increase in heat tolerance of heat-adapted strains compared to the wild type (WT) strains. In particular, the survival ratio of basically low heat-tolerant strain increased even more. In addition, the strains with improved heat tolerance acquired cross protection, which improved their survival ratio in acid, bile salts and osmotic conditions. A relation between heat tolerance and membrane fatty acid composition was identified. As a result of heat adaptation, the ratio of unsaturated to saturated fatty acids (UFA/SFA) and C18:1 relative concentration were decreased. C6:0 in only heat-adapted strains and C22:0 in only the naturally high heat tolerant strain were detected. These results support the hypothesis, that the consequent increase of SFA ratio is a cellular response to environmental stresses such as high temperatures, and it is able to protect the cells from acid, bile salts and osmotic conditions via cross protection. This study demonstrated that the increase in heat tolerance can be utilized as a mean to improve bacterial tolerance against various environmental stresses.

Characterization of Selected Lactobacillus Strains for Use as Probiotics

  • Song, Minyu;Yun, Bohyun;Moon, Jae-Hak;Park, Dong-June;Lim, Kwangsei;Oh, Sejong
    • Food Science of Animal Resources
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    • v.35 no.4
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    • pp.551-556
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    • 2015
  • The aim of this study was to evaluate the functional properties of lactic acid bacteria from various sources and to identify strains for use as probiotics. Ten Lactobacillus strains were selected and their properties such as bile tolerance, acid resistance, cholesterol assimilation activity, and adherence to HT-29 cells were assessed to determine their potential as probiotics. Lactobacillus sp. JNU 8829, L. casei MB3, L. sakei MA9, L. sakei CH8, and L. acidophilus M23 were found to show full tolerance to the 0.3% bile acid. All strains without L. acidophilus M23 were the most acid-tolerant strains. After incubating the strains at pH 2.5 for 2 h, their viability decreased by 3 Log cells. Some strains survived at pH 2.5 in the presence of pepsin and 0.3% bile acid. Lactobacillus sp. JNU 8829, L. acidophilus KU41, L. acidophilus M23, L. fermentum NS2, L. plantarum M13, and L. plantarum NS3 were found to reduce cholesterol levels by >50% in vitro. In the adhesion assay, Lactobacillus sp. JNU 8829, L. casei MB3, L. sakei MA9, and L. sakei CH8 showed higher adhesion activities after 2 h of co-incubation with the intestinal cells. The results of this comprehensive analysis shows that this new probiotic strain named, Lactobacillus sp. JNU 8829 could be a promising candidate for dairy products.

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

  • Ryu, Ji-Sook;Han, Sun-Kyung;Shin, Myeong-Soo;Lee, Wan-Kyu
    • Korean Journal of Veterinary Service
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    • v.32 no.1
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    • pp.33-41
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    • 2009
  • In order to develop probiotic strain for pigs, Lactobacillus spp. (527 isolates), Streptococcus spp. (95 isolates) and Bifidobacterium spp. (25 isolates) were isolated from the feces of 35 pigs. These isolates were tested through in vitro experiment such as acid tolerance at pH 2.0 (Lactobacillus spp. and Streptococcus spp.) or pH 3.0 (Bifidobacterium spp.), bile tolerance in MRS broth containing 0.3% (w/v) Oxgall, heat resistance at $70^{\circ}C$ and $80^{\circ}C$ for 5 min, antibiotic resistance, antimicrobial activity against pathogenic bacteria and Caco-2 cell adherence assay. Finally ten most superior strain (5 Lactobacillus spp. strain, 3 Bifidobacterium spp. strain and 2 Streptococcus spp. strain) were selected as potential candidate for probiotic use in pig industry. It could be used as an alternative to antibiotics in feed additives.

Characterization of Lactobacillus cellobiosus D37 Isolated from Soybean Paste as a Probiotic with Anti-Cancer and Antimicrobial Properties

  • Lim, Sung-Mee;Lee, Goon-Ja;Park, Sun-Mee;Ahn, Dong-Hyun;Im, Dong-Soon
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.792-798
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    • 2006
  • The probiotic characteristics of a total of 137 lactic acid bacterial strains isolated from soybean paste were investigated. Among those tested, the D37 strain was selected as a probiotic bacteria due to its acid and bile tolerance, and its strong anti-cancer and antibacterial activities. The D37 strain showed highly stable viability at acidic pH for 2 hr, and was very stable in 10% bovine bile. The viability of human colon cancer HT-29 cells was inhibited more than 60% at a $200\;{\mu}/mL$ concentration of D37 cell-free culture supernatant, and the degree of inhibition was concentration-dependent. The D37 strain showed a wide range of antibacterial activities against food-borne pathogenic bacteria such as Escherichia coli O157, Listeria spp., Vibrio spp., Salmonella spp., and Staphylococcus aureus. According to phenotypic characteristics and the utilization of various sugars, the D37 strain was identified as Lactobacillus cellobiosus.

Functional Properties of Yogurt Containing Specific Peptides derived from Whey Proteins

  • Won, Ji-Young;Kim, Hong-Soek;Jang, Jin-Ah;Kim, Cheol-Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.4
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    • pp.249-254
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    • 2017
  • The purpose of this study was to investigate the acid tolerance, bile acid tolerance, and fermentation activity of lactic acid bacteria isolated from Kimchi in the presence of hydrolysates of whey protein concentrate. Kimchi isolates DK109, DK119, DK121, DK128, DK211, DK212, and DK215, which were identified as Lactobacillus sp., and L. casei DK128 showed the highest acid and bile acid tolerance. To produce whey hydrolysates, enzymes were added to a 10% (w/v) whey protein concentrate (WPC) solution at 1:50 (w/v, protein). The viabilities of the DK strains were determined in the presence of low pH and bile salts. Then, yogurt was produced via fermentation with L. casei DK128, an isolate from Kimchi, in the presence of the following additives: CPP, WPC, and WPC hydrolysates (WPCH) generated by alcalase (A) or neutrase (N). The produced yogurts were subjected to various analyses, including viable cell counts (CFU/mL), pH, titratable activity, and sensory testing. After 8 h of fermentation, the pH and titratable activity values of all test samples were 4.2 and 0.9, respectively. The viable counts of LAB were $3.49{\times}10^8$, $5.72{\times}10^8$, $7.01{\times}10^8$, and $6.97{\times}10^8$, for the Control, CPP, A, and N samples, respectively. These results suggest that whey proteins have potential as dietary supplements in functional foods and that WPCH could be used in yogurt as a low-cost alternative to CPP.

Characterization of Lactobacillus acidophilus Isolated from Piglets and Chicken

  • Ahn, Y.T.;Lim, K.L.;Ryu, J.C.;Kang, D.K.;Ham, J.S.;Jang, Y.H.;Kim, H.U.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1790-1797
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    • 2002
  • Lactic acid bacteria were isolated from piglets and chicken and characterized. Lactic acid bacteria showing resistance to low pH and bile, adhesion to intestinal epithelium cells, and the inhibition of Escherichia coli and Salmonella spp. were identified as Lactobacillus acidophilus. L. acidophilus PF01 survived for 2 h in MRS broth adjusted to pH 2. L. acidophilus CF07 was less resistant than L. acidophilus PF01 to pH 2, but survived at pH 2.5 for 2 h. Both of isolates were able to grow in MRS broth containing 0.3% (w/v) bile, with L. acidophilus CF07 being more tolerant to bile than L. acidophilus PF01. L. acidophilus PF01 and CF07 adhered specifically to the duodenal and jejunal epithelium cells of piglet, and the cecal and duodenal epithelium cells of chicken, respectively. Both of isolates did not adhere to the epithelium cells of the various animal intestines from which they were isolated. When L. acidophilus was cultured with E. coli and Salmonella spp. in MRS broth, MRS broth containing 2% skim milk powder or modified tryptic soy broth at $37^{\circ}C$, L. acidophilus PF01 and CF07 inhibited the growths of E. coli K88 and K99, and S. enteritidis and S. typhimurium, respectively. Both of isolates were found to possess the essential characteristics of probiotic lactic acid bacteria for piglet and chicken.

Biochemical Characteristics of Lactobacillus acidophilus Isolated from a Breast-Fed Infant (모유 섭취 신생아 유래 Lactobacillus acidophilus의 생리적 특성)

  • Hong, Sung-Moon;So, Byung-Chun;Yoon, Seumg-Won;Kim, Cheol-Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.1
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    • pp.45-53
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    • 2012
  • Lactobacillus acidophilus isolated from the feces of a 7-day-old breast-fed infant was characterized to examine the scope of its commercial use. Forty-three Lactobacillus strains, which could grow at pH 5.5, were isolated. From these Lactobacillus isolates, 14 Lactobacillus strains were selected, which demonstrated more than 80% viability and homofermentative lactic fermentation. Finally, 9 L. acidophilus strains (NB 201~NB 209) were identified as candidate strains based upon biochemical properties, carbohydrate utilization, and cellular fatty acid composition. L. acidophilus isolates demonstrated a survival rate of more than 80% when exposed to pH 2.5 for 2 h. In particular, L. acidophilus NB 204 showed a strong acid tolerance, with a 71% survival rate even at pH 2.0. L. acidophilus isolates also manifested high bile acid tolerance; more than 87% of the cells survived on agar containing 1% bile extract, except for L. acidophilus NB 206, which showed a 73% survival rate. All L. acidophilus isolates were confirmed to have proteolytic activity; L. acidophilus NB 204 and NB 209 yielded higher levels of TCA-soluble peptides and free amino acids. The ${\beta}$-galactosidase activity of the L. acidophilus isolates was in the range of 1.97~2.45 units/mL.

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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.

Characteristics of Paenibacillus sp. BCNU 5016 as a Novel Probiotic (신규 프로바이오틱스로서 Peanibacillus sp. BCNU 5016의 특성)

  • Choi, Hye Jung;Kim, Dong Wan;Joo, Woo Hong
    • Journal of Life Science
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    • v.24 no.2
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    • pp.161-166
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    • 2014
  • We screened Paenibacillus sp. strains from Jeotgal, a Korean salted and fermented fish product, for use as a novel probiotic. Among these Paenibacillus sp. isolates, BCNU 5016 was a typical Paenibacillus sp. strain that showed gram-positive, gelatinase-negative, and urease negative activity. On the basis of 16S rDNA sequence comparisons, BCNU 5016 was most closely related phylogenetically to P. polymyxa. When Paenibacillus sp. BCNU 5016 was subjected to the acid tolerance test, this strain showed 91.89% survival after 3 h culture at pH 2.5. Paenibacillus sp. BCNU 5016 also showed excellent bile acid tolerance. Furthermore, its auto-aggregation, coaggregation, and hydrophobic capacities suggest that BCNU 5016 had the capacity to adhere well to the intestinal tract. We conclude that Paenibacillus sp. BCNU 5016 has excellent potential as a probiotic.

Characterization and Immunomodulation Activity of Lactobacillus sakei L2 and L8 Isolated from Chicken Cecum (닭의 맹장으로부터 분리한 Lactobacillus sakei L2와 L8의 특성 및 면역활성)

  • Sim, Insuk;Park, Keun-Tae;Lim, Young-Hee
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.201-207
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    • 2016
  • The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.