• Title/Summary/Keyword: L. acidophilus

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Molecular Cloning and Characterization of a Bile Salt Hydrolase from Lactobacillus acidophilus PF01

  • Oh, Hae-Keun;Lee, Ji-Yoon;Lim, Soo-Jin;Kim, Min-Jeong;Kim, Geun-Bae;Kim, Jung-Hoan;Hong, Soon-Kwang;Kang, Dae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.449-456
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    • 2008
  • Phenotypic screening for bile salt hydrolase (BSH) activity was performed on Lactobacillus acidophilus PF01 isolated from piglet feces. A gene encoding BSH was identified and cloned from the genomic library of L. acidophilus PF01. The bsh gene and surrounding regions were characterized by nucleotide sequence analysis and were found to contain a single open reading frame (ORF) of 951 nucleotides encoding a 316 amino acid protein. The potential bsh promoter region was located upstream of the start codon. The protein deduced from the complete ORF had high similarity with other BSHs, and four amino acid motifs located around the active site, FGRNXD, AGLNF, VLTNXP, and GXGXGXXGXPGD, were highly conserved. The bsh gene was cloned into the pET21b expression vector and expressed in Escherichia coli BLR(DE3) by induction with 0.1mM of isopropylthiogalactopyranoside. The BSH enzyme was purified with apparent homogeneity using a $Ni^{2+}$-NTA agarose column and characterized. The overexpressed recombinant BSH enzyme of L. acidophilus PF01 exhibited hydrolase activity against tauroconjugated bile salts, but not glycoconjugated bile salts. It showed the highest activity against taurocholic acid. The maximum BSH activity occurred at approximately $40^{\circ}C$. The enzyme maintained approximately 70% of its maximum activity even at $60^{\circ}C$, whereas its activity rapidly decreased at below $37^{\circ}C$. The optimum pH was 6, and BSH activity was rapidly inactivated below pH 5 and above pH 7.

Characterization of a Lactobacillus acidophilus Strain Isolated from Korean Infant Feces and Cloning of Surface Layer Protein Gene slp and Its Expression in Escherichia coli (유아 분변에서 분리한 Lactobacillus acidophilus의 특성 및 표면 단백질 유전자 클로닝과 대장균 내에서의 발현)

  • Park, Myeong-Soo;Ji, Geun-Eog;You, Kwan-Hee;Lee, Si-Kyung;Jeong, Won-Seok;Kim, Jin-Hyung;Jo, Myoung-Hwan;Kim, Soo-Young
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.352-356
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    • 2007
  • A Lactobacillus sp. has been isolated from infant feces and characterized according to its physiological properties and identified as Lactobacillus acidophilus KLA1012. A gene coding surface layer protein (SLP) has been cloned and the sequence has been determined. The nucleotide sequence of slpA was 1,338 bp in size and was identical to that of L. acidophilus ATCC 4356 (100%). Amino acid sequence of SLP-A was deduced from the nucleotide sequence and it had signal sequence at N-terminal, consisting of positively charged amino acid mainly lysine. slpA was cloned and heterologously expressed in E. coli M15 and the 45.2 kDa surface-layer protein band was examined by SDS-PAGE and confirmed by Western blotting using polyclonal antibody against L. acidophilus KLA 1012 SLP-A protein.

Lactobacillus acidophilus Strain Suppresses the Transcription of Proinflammatory-Related Factors in Human HT-29 Cells

  • Chen, Kun;Liang, Nailong;Luo, Xuegang;Zhang, Tong-Cun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.64-68
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    • 2013
  • Previous studies have shown that lactic acid bacteria can inhibit inflammatory responses, but the mechanisms are very little known. In this study, transaction and expression of three proinflammatory factors, iNOS, PTGS-2, and IL8, which are closely related to the inflammatory response, were investigated by luciferase reporter assay and RTPCR in HT-29 cells treated by Lactobacillus acidophilus. The results showed that the live L. acidophilus sharply down-regulated the transcription of these three genes. Because there was a NF-${\kappa}B$ binding site located at -265 bp, -225 bp, and -95 bp upstream of the iNOS, PTGS-2, and IL8 promoters, respectively, we further addressed the effects of NF-${\kappa}B$ on transaction of the three promoters by cotransfection. As was expected, NF-${\kappa}Bs$ remarkably upregulated the activity of the reporter gene and, no effect of NF-${\kappa}B$s on IL-8 promoter transaction was found after NF-${\kappa}B$ binding site mutation of the IL8 promoter in HT-29 cells. In conclusion, the live L. acidophilus decreased the transcriptional activity of NF-${\kappa}B$ and, in turn, inhibited the transaction of NF-${\kappa}B$ on the three proinflammatory factors mentioned above.

Purification, Bacteriolytic Action and Plasmid Isolation of Acidocin 4A Produced by Lactobacillus acidophilus GP4A (Lactobacillus acidophilus GP4A 박테리오신의 정제, Bacteriolytic 작용 및 생산 관련 Plasmid의 선별)

  • Han, Gyeong-Sik;Jeon, U-Min;Kim, Yeong-Hun;Kim, Se-Heon
    • Journal of Dairy Science and Biotechnology
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    • v.21 no.2
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    • pp.114-119
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    • 2003
  • Acidocin 4A produced by Lactobacillus acidophilus GP4A was purified to homogeneity by ammonium sulfate precipitation and sequential chromatographies containing Octyl sepharose CL-4B column, $C_{18}$ Sep-Pak Cartridge, $C_{18}$ RP HPLC and HPLC gel filtration. Tricine SDS-PACE resulted in a single band with estimated molecular mass of 4.1 kDa corresponding to the polypeptide weight marker. Electron microscopy of acidocin-treated indicator cells(L. delbrueckii subsp. lactis ATCC 4797) confirmed that acidocin 4A presented bacteriolytic effect, resulting in cell lysis. Curing trial using ethidium bromide (EtBr) was carried out to examine whether acidocin 4A determinant was encoded either by chromosome or on plasmid. The plasmid designated as pLA4A, being about 20 kb in size, was responsible for acidocin 4A production and immunity to host cells.

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Effects of cholesterol and Lactobacillus acidophilus on testicular function

  • Ciftci, Gulay;Tuna, Elif
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.3
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    • pp.229-235
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    • 2021
  • Objective: In this study, the effects of Lactobacillus acidophilus on testosterone (TES), follicle-stimulating hormone (FSH), luteinizing hormone (LH), androgen-binding protein (ABP), factor-associated apoptosis (FAS), and total cholesterol (TC), as well as histopathological changes, were investigated in male rats fed a high-cholesterol diet. Methods: The study included three groups. The control (C) group was fed standard-diet for 8 weeks. The hypercholesterolemia (HC) group was fed a 2% cholesterol-diet for 8 weeks. The therapeutic group (HCL) was fed a 2% cholesterol-diet for 8 weeks and administered L. acidophilus for the last 4 weeks. FSH, TES, and FAS levels in testicular tissue were determined using an enzyme-linked immunosorbent assay (ELISA), while another sample was examined histopathologically. LH and ABP levels were determined using ELISA, and serum TC levels were assessed via an autoanalyzer. Results: In the HC group, the TC levels were significantly higher and the LH levels were lower (p<0.05) than in the C group. The ABP levels were lower (p>0.05). In the HCL group, the LH and ABP levels were higher (p>0.05) and the TC level significantly lower (p<0.05) than in the HC group. The TES and FSH levels were lower, and the FAS levels were higher, in the HC than in the C group (p<0.05). In the HCL group, levels of all three resembled control levels. Histologically, in the testicular tissue of the HC group, the cells in the tubular wall exhibited atrophy, vacuolization, and reduced wall structure integrity. However, in the HCL group, these deteriorations were largely reversed. Conclusion: Supplementary dietary administration of an L. acidophilus to hypercholesterolemic male rats positively impacted testicular tissue and male fertility hormone levels.

The Effect of Various Culture Conditions on Lactic Acid Bacteria Count (배양조건에 따른 유산균수 비교시험)

  • Kim, Eun-Ah;Kwak, Hae-Soo;Park, Joung-Nam
    • Korean Journal of Food Science and Technology
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    • v.22 no.4
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    • pp.421-425
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    • 1990
  • This study was carried out to compare lactic acid bacteria count of liquid type yogurts with various experimental conditions during shelf-life period. The conditions were media(BCP and Elliker agar), incubation conditions(aerobic and anaerobic), dilution waters(saline and phosphate buffer) and dilution methods(10 and 100 times). All of the samples were incubated at $37^{\circ}C$ for 72 hrs. In the case of counting L. acidophilus as a yogurt starter culture, there were differences on dilution waters and incubation conditions, but were no difference on media and dilution methods. In the case of counting L jugurti and mixed strain with L. acidophilus and L. casei, there were differences on media, incubation conditions and dilution waters, but was no difference on dilution methods. For L. casei in the yogurt, media and dilution methods were shown slightly different viable cell count but incubation conditions were not shown difference. In the case of counting L. bulgaricus, there were differences on media, incubation conditions and dilution methods, but was no difference on dilution waters. Therefore, the measurment of lactic acid bacteria count may be effective if preferred experimental conditions are selected for different types of strain.

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Lactobacillus acidophilus KFCC12731와 Kluyveromyces fragilis KFCC35457의 혼합배양을 이용한 대두유의 젖산발효조건

  • 류인석;박정길;유주현
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.531.1-531
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    • 1986
  • L. acidophilus를 대두유에 젖산발효 시킬 때 단독배양과 효모인 K. fragilis와의 혼합배양에 대하여 검토하였다. 젖산의 생성량은 젖산균 단독배양보다는 효모와 혼합배양 하는 것이 2.6배나 많았으며 L. acidophilus를 단독배양 하였을 경우 균의 생육속도와 PH 강하속도는 K. fragilis를 단독배양한 것에 비교하여 늦었다. 그러나 혼합배양 함으로써 생육속도와 PH 강하속도는 K, fragilis를 단독배양한 것보다 빨랐다. 단독 또는 혼합젖산발효에 미치는 온도의 영향을 조사한 결과는 L. acidophilus.는 37$^{\circ}C$, K. frahilis는 3$0^{\circ}C$, 두 균의 혼합배양시는 35-37$^{\circ}C$ 범위에서 산생성이 가장 좋았다. 혼합배양 시 검토한 젖산발효 최적조건은 L. acidophilus와 K. fragilis의 접종비율이 1:5, sucrose 0.5-1.0% SKIMMILK 1.5%를 각각 첨 가한 것이 좋았다.

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Stydies on the Selective Screening Method of Bifidobacteria Used in Yoghurt (호상 요구르트 제품에 이용되는 Bifidobacteria 의 선택적 검출 방법)

  • 박희경;허태련
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.214-219
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    • 1995
  • In fermentation of dairy products, bifidobacteria is used in conjunction with other lactic acid bacteria, such as L. acidophilus, L. bulgaricus and S. thermophilus, rendering the enumeration of bifidobacteria difficult. In order to develop optimum conditions for selective enumeration of bifidobacteria, we examined MIC of several antibiotics against various bifidobacteria and other lactic acid bacteria. The growth of L. acidophilus, L. bulgaricus and S. thermophilus were inhibited by lithium chloride at the concentration of less than 4 mg/ml, whereas growth inhibition of bifidobacteria occurred at concentrations over 6-10 mg/ml. Tetracycline and chloramphenicol were also found to selectively inhibit growth of other lactic acid bacteria at the concentration of 1-3 $\mu$g/ml. Addition of 6 mg/ml lithium chloride, 1 $\mu$g/ml, tetracycline or 3 $\mu$g/ml chloramphenicol to medium was found to be optimal for selective enumeration of bifidobacteria. By using these three inhibitory chemicals in the TPY medium, higher number of bifidobacteria were selectively isolated than with NPNL agar and LP agar.

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Antilisterial Bacteriocin from Lactobacillus rhamnosus CJNU 0519 Presenting a Narrow Antimicrobial Spectrum

  • Jeong, Ye-Jin;Moon, Gi-Seong
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.137-142
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    • 2015
  • A lactic acid bacterium presenting antimicrobial activity against a Lactobacillus acidophilus strain used for eradication of acid inhibition was isolated from a natural cheese. The 16S rRNA gene sequence of the isolate best matched with a strain of L. rhamnosus and was designated L. rhamnosus CJNU 0519. The antimicrobial activity of the partially purified bacteriocin of CJNU 0519 was abolished when treated with a protease, indicating the protein nature of the bacteriocin. The partially purified bacteriocin (rhamnocin 519) displayed a narrow antimicrobial activity against L. acidophilus, Listeria monocytogenes, and Staphylococcus aureus among several tested bacterial and yeast strains. Rhamnocin 519 in particular showed strong bactericidal action against L. monocytogenes.

Screening of Radio-resistant Lactic Acid Bacteria

  • Hwang, E-Nam;Kang, Sang-Mo;Kim, Jae-Kyung;Lee, Ju-Woon;Park, Jong-Heum
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.335-340
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    • 2013
  • This study screened for radio-resistant strains lactic acid bacteria (LAB) by evaluating their capability to survive exposure to ionizing radiation. Ten strains of LAB - Lactobacillus bulgaricus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus delbruekii, Lactococcus lactis, Streptococcus thermophilus, Bifidobacterium breve, and Pediocuccos pentosaceus - were selected and subcultuted twice. The LAB was then further cultured for 3 d at $37^{\circ}C$ to reach 7-10 Log colony-forming units (CFU)/mL prior to irradiation and immediately exposed to gamma rays or electron beams with absorbed doses of 0, 1, 2, 3, 4, 5, 6, 8, and 10 kGy. Gamma irradiation gradually decreased the number of the tested viable LAB, and the effect was irradiation dose dependent. A similar effect was found in electron beam-irradiated LAB. Radiation sensitivity of LAB was calculated as $D_{10}$ values, which ranged from 0.26 kGy to 0.9 kGy and 0.5 kGy to 1.44 kGy with exposure to gamma and electron beam irradiation, respectively, in all tested LAB. L. acidophilus was the most resistant to gamma and electron beam irradiation, with $D_{10}$ values of 0.9 kGy and 1.44 kGy, respectively. These results suggest that L. acidophilus might be suitable for the preparation of probiotics as direct-fed microbes for astronauts in extreme space environments.