• Title, Summary, Keyword: Lactobacillus casei

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Transformation of Escherichia coli-Lactobacillus casei Shuttle Vector by Electroporation (Electroporation에 의한 Escherichia coli-Lactobacillus casei 셔틀 벡터의 형질전환)

  • 홍성희
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.109-111
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    • 2000
  • A 3.5 kb plasmid from Lactobacillus. casei ssp. cosei NCIB 4114 was isolated and E. cali-L. casei shuttle vectors were constructed containing this plasmid. Transformation by electroporation was successful with all the plasmids constructed. Optimized condition for the electroporation was established with efficiency level of $2{\times}10^5$ transformants per $\mu$g of vector DNA. Successful introduction of those shuttle vectors enable to these vectors as food grade vector for lactic acid bacteria.

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Studies on the Protoplast Fusion between Lactobacillus casei and Lactobacillus delbrueckii (Lactobacillus casei와 Lactobacillus delbrueckii간의 Protoplast 융합에 관한 연구)

  • 전홍기;김미경;백형석
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.6-13
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    • 1992
  • - Protoplast fusion between lincomycin resistant Lactobacillus casei KCTC 1121 and rifarnpicin resistant Lactobacillus delbrueckii JK-414 was attempted to obtain the improved strains. Protoplasts of L. casei and L. delbrueckii were produced by mutanolysin digestion at $42^{\circ}C$ for 15 min. L. casei cells were converted to protoplasts by treating with 5 $\mu g$ / m l of mutanolysin in 20 mM HEPES buffer (pH 7.0) containing 0.75 M sucrose at the middle logarithmic growth phase. In case of L. delbrueckii 1.0 M sucrose was used osmotic stabilizer. Regeneration of protoplast in both strains was efficiently accomplished on the regeneration medium containing 10% sucrose, 6 mM $MgC1_2, 6 mM CaC1_2$, and 2.5% gelatin. Protoplast fusion between L. casei and L. delbrueckii was carried out in the presence of 40% of PEG 4,000. The frequency of protoplast fusion was found to be about $3.2\times 10^4$. Acid production of L. casei was better than that of L. delbrueckii. Among fusants, F23 and F35 exhibited excellent lactic acid production. F23 and F24 exhibited the improved proteolysis compared to that of the parent strains and they had twice as much as DNA content of the parents.

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Analysis of Lactobacillus casei and Mutant Strains by Polymerase Chain Reaction (Polymerase Chain Reaction에 의한 Lactobacillus casei 및 돌연변이 균주들의 비교 분석)

  • Nam, Jin-Sik;Lee, Jeong-Jun;Shin, Myeong-Su;Na, Seog-Hwan;Baek, Young-Jin;Yoo, Min
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.577-583
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    • 1994
  • To classify Lactobacillus casei strains on the basis of difference in their chromosomal DNA sequence, we have performed polymerase chain reactions on their chromosomal DNA by using random primers, and followed by analyzing randomly amplified polymorphic DNA fragments. We also developed a mini-preparative method to isolate PCR-grade chromosomal DNA from Lacto- bacillus casei strains within 3 hours. Based on RAPD pattems by polymerase chain reactions with degenerated random primers, 4 Lactobacillus casei strains and 2 mutant strains were successfully discriminated. Results were very sensitive, strain-specific and reproducible. It was also reliable. These results suggest that RAPD may be applied efficiently for the identification of several Lactoba- cillus casei strains.

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Expression of Bacillus licheniformis $\alpha$-amylase Gene in Lactobacillus casei Strains

  • Kim, Jeong Hwan;Sung Hee Woo
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.257-263
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    • 1995
  • As a first step for developing Lactobacillus strains capable of fermenting starch directly, the $\alpha$-amylase gene (amyL) from Bacillus licheniformis (Kim et al., 1988. Kor. J. Appl. Microbiol. Bioeng. 16: 369-373) was introduced into Lactobacillus casei strains and the level of $\alpha$-amylase expression in transformants was examined. 3 kb EcoRI fragments encompassing amyL were subcloned into the suitable lactococcal cloning vectors (pSA3, pMG36e, and p1L2530) and then recombinant plasmids were introduced into E. coli and L. casei strains by electroporation. Only one recombinant plasmid, $pIL2530\alpha$ was able to transform few L. casei strains tested at low efficiencies. The transformation efficiencies with the plasmid into L. casei YIT 9018 and L. casei A Tee 4646 were less than $10^2/\mu$ g pIL2530\alpha$. The level of amylase activities in L. casei was five to ten-fold lower than that in E. coli cells. $p1L2530\alpha$ was stably maintained in Lactobacillus strains in the presence of Em (5 $\mu $g/ml) but without antibiotic selection, it was unstable so more than 95$%$ of cells lost plasmids after a week of daily subculturing.

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Orally administered Lactobacillus casei exhibited several probiotic properties in artificially suckling rabbits

  • Shen, Xue Mei;Cui, Hong Xiao;Xu, Xiu Rong
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.8
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    • pp.1352-1359
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    • 2020
  • Objective: Lactobacilli in rabbit intestine is rare and its function in rabbit gut health is not fully understood. The present study aimed to evaluate in vivo the probiotic potential of Lactobacillus casei for suckling rabbits. Methods: Two healthy 5-day-old suckling rabbits with similar weights from each of 12 New Zealand White litters were selected and disturbed to control group and treatment group. All rabbits were artificially fed. The treatment group had been supplemented with live Lactobacillus casei in the milk from the beginning of the trial to 13 days of age. At 15 days of age, healthy paired rabbits were slaughtered to collect intestinal samples. Results: i) Oral administration of Lactobacillus casei significantly increased the proportion of Lactobacilli in the total intestinal bacteria (p<0.01) and obviously reduced that of Escherichia-Shigella (p<0.01); ii) treatment increased the length of vermiform appendix (p<0.05); iii) a higher percentage of degranulated paneth cells was observed in the duodenum and jejunum when rabbits administered with Lactobacillus casei (p<0.01); and iv) the expression of toll-like receptor 9, lysozyme (LYZ), and defensin-7-like (DEFEN) in the duodenum and jejunum was stimulated by supplemented Lactobacillus casei (p<0.05). Conclusion: Orally administered Lactobacillus casei could increase the abundance of intestinal Lactobacilli, decrease the relative abundance of intestinal Escherichia-Shigella, promote the growth of appendix vermiform, stimulate the degranulation of paneth cells and induce the expression of DEFEN and LYS. The results of the present study implied that Lactobacillus casei exhibited probiotic potential for suckling rabbits.

Suppressive Effect of Lactic Acid Bacteria on the Toxicity of Bisphenol A in Rats

  • Yoo, Min;Min, Byung-Tae
    • Biomedical Science Letters
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    • v.7 no.1
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    • pp.27-30
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    • 2001
  • We have examined if lactic acid bacteria could suppress the toxic effect of bisphenol A. Lactobacillus casei YA-70 was chosen as representative. Thirty rats were divided into two groups (immature and mature) according to the weight. Each group was divided again into the control group (only alcohol treatment), bisphenol A treated group, and bisphenol A plus Lactobacillus casei YA-70 treated group. When 500 ppm of bisphenol A was fed everyday, 83% of immature group and 50% of mature group died within 3 weeks. Their internal organs, mainly livers and lungs, were changed in color and severely damaged. In the intestine of 5 ppm-fed group tumor-like nodules were observed. However, their number and size were markedly decreased when Lactobacillus casei YA-70 was supplemented in diet. This study strongly indicates that Lactobacillus casei YA-70 might play an important role to suppress the toxic effect of endocrine disruptor.

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Construction of Recombinant Lactobacillus casei Strains Using Splicing by Overlap Extension

  • Jeong, Do-Won;Lee, Jong-Hoon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1953-1957
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    • 2008
  • Recombinant Lactobacillus strains have been constructed using gene splicing by overlap extension (SOE). Primers were designed of which one end of an amplified product contained complementary sequences for an end of other amplified fragment. For efficient matching, we used an asymmetric PCR step that was effective at generating an excess of strands that would anneal in the final PCR. CP12, a recombinant fragment consisting of the integrase gene and attachment site of the bacteriophage A2, was constructed and inserted into the genome of Lactobacillus casei ATCC 393, yielding Lb. casei ATCC 393::XCP12. Another recombinant Lb. casei strain was constructed, where the egfp gene was a part of the construction. The EGFP produced from Lb. casei ATCC 393::XCEGFP14 was detected by Western blot hybridization. This simple and widely applicable approach has significant advantages over standard recombinant DNA techniques for Lactobacillus species.

Beneficial Effects of Lactobacillus casei ATCC 334 on Halitosis Induced by Periodontopathogens

  • Lee, Ki-Ho;Baek, Dong-Heon
    • International Journal of Oral Biology
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    • v.39 no.1
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    • pp.35-40
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    • 2014
  • Halitosis is caused by consumption of certain foods or drinks and production of volatile sulfur compounds (VSCs) by periodontopathogens. VSCs-related halitosis is not easily removed using mechanical or chemical therapies such as dental floss, plaque control and mouth rinse. Lactobacillus are known to be probiotics and stimulate immune systems of human. Furthermore, L. casei ATCC 334 and L. rhamnosus GG have an effect on protection of dental caries in vitro studies. The aim of this study was to investigate effect of Lactobacillus on halitosis by Fusobacterium nucleatum- and Porphyromonas gingivalis-producing VSCs and to analyze inhibitory mechanism. The periodontopathogens were cultivated in the presence or the absence Lactobacillus, and the level of VSCs was measured by gas chromatograph. For analysis of inhibitory mechanisms, the susceptibility assay of the spent culture medium of Lactobacillus against F. nucleatum and P. gingivalis was investigated. Also, the spent culture medium of Lactobacillus and periodontopathogens were mixed, and the emission of VSCs from the spent culture medium was measured by gas chromatograph. L. casei and L. rhamnosus significantly reduced production of VSCs. L. casei and L. rhamnosus exhibited strong antibacterial activity against F. nucleatum and P. gingivalis. The spent culture medium of L. casei inhibited to emit gaseous hydrogen sulfide, methyl mercaptan and dimethyl sulfide from the spent culture medium of periodontopathogens. However, the spent medium of L. rhamnosus repressed only dimethyl sulfide. L. casei ATCC 334 may improve halitosis by growth inhibition of periodontopathogens and reduction of VSCs emission.

Characterization of the cured mutants of Lactobacillus casei (Lactobacillus casei YIT 9018의 Mutants의 특성)

  • 유선이;강현삼
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.184-189
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    • 1985
  • The cured mutant strains (CW, CSM, CEM, CAM) we obtained from Lactobacillus casei YIT 9018 spontaneously and by ethidium bromide treatments. The lactose fermenting ability of those mutants was tested and plasmid was found to have some functions in lactose metabolism. Plasmid (PLC) has been detected in Lactobacillus casei YIT 9018, but has not been detected in mutants CSM, CEM, CAM. These plasmid curred mutants showed a decrease in the ability to produce lactic acid from lactose.

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Properties of the Fusants of Lactobacillus acidophilus 88 and Lactobacillus casei subsp. casei KCTC 1121

  • Jo, Young-Bae;Heo, Kyeong;Kim, Sung-Koo;Baik, Hyung-Suk;Jun, Hong-Ki
    • Journal of Microbiology and Biotechnology
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    • v.7 no.1
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    • pp.25-31
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    • 1997
  • Protoplast fusion between L. casei KCTC 1121 and L. acidophilus 88 was attempted to obtain improved strains. The fusants produced a bacteriocin against indicator strains, making a smaller inhibition zone compared to that of L. acidophilus 88. After culturing for 2 months on selective medium, the selected fusants were still stable without segregation. Fusants showed higher lipase activity compared to those of the two parent strains. Fusant No.4, 11, and 15 exhibited excellent lactic acid productivity. Fusant No.4 and 15 exhibited improved proteolysis ability compared to the two parent strains. Whereas L. casei possessed both ${\beta}-galactosidase$ and $phospho-{\beta}-galactosidase$ activities, and L. acidophilus 88 had only ${\beta}-galactosidase$ activity, the fusants had both the intermediate enzyme activities. Cell size of the fusants was greater than that of the parents.

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