• Title/Summary/Keyword: L. lactis

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In situ Delivery of Therapeutic Proteins by Recombinant Lactococcus lactis

  • Steidler, Lothar;Neirynck, Sabine
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.63-72
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    • 2003
  • Chronic inflammatory bowel disease (IBD) such as Crohn's disease or ulcerative colitis, affects around 2 in every 1000 individuals in western countries and its incidence, particularly amongst children, is increasing. IBD shows extreme morbidity with impact on all aspects of quality of life. If left untreated, IBD can lead to death. Conventional treatment of IBD involves powerful immunosuppressive chemotherapies and surgical intervention. Long-term anti-inflammatory medication is required and so patients are often subject to a spectrum of unpleasant side effects. Interleukin-10 (IL-10) is a cytokine that acts to suppress inflammation. When however administered by injection, the high levels of IL-10 that are distributed throughout the body also lead to side effects. Lactococcus lactis can be genetically engineered to secrete biologically active cytokines. When applied to the mucosa, these L. lactis can actively deliver such cytokines. By use of this principle we developed a new therapeutic approach for IBD. Administration of L. lactis that secretes murine IL-10 cures and prevents IBD in mice. The use of the engineered L. lactis gets around the problem of delivering IL-10, allowing dramatic reduction of the effective dose. A sincere concern exists about the possible dangers of uncontrolled, deliberate release of genetically modified microorganisms, such as could occur following application in healthcare. We engaged in the establishment of adequate means for biological growth control of engineered L. lactis by targeted gene exchange between thyA and hIL-10.

Cloning and Expression of hpaA Gene of Korean Strain Helicobacter pylori K51 in Oral Vaccine Delivery Vehicle Lactococcus lactis subsp. lactis MG1363

  • Kim Su-Jung;Jun Do-Youn;Yang Chae-Ha;Kim Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.318-324
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    • 2006
  • In order to develop an oral vaccine to prevent H. pylori infection, we have expressed the hpaA gene of H. pylori K51 isolated from Korean patients, encoding 29-kDa HpaA that is known to be localized on the cell surface and flagella sheath, in a live delivery vector system, Lactococcus lactis. The hpaA gene, amplified by PCR using the genomic DNA of H. pylori K51, was cloned in the pGEX-2T vector, and the DNA sequence analysis revealed that the hpaA gene of H. pylori K51 had 99.7% and 94.8% identity with individual hpaA genes of the H. pylori 26695 strain (U.K) and the J99 strain (U.S.A). A polyclonal anti-HpaA antibody was raised in rats using GST-HpaA fusion protein as the antigen. The hpaA gene was inserted in an E. coli-L. lactis-shuttle vector (pMG36e) to express in L. lactis. Western blot analysis showed that the expression level of HpaA in the L. lactis transformant remained constant from the exponential phase to the stationary phase, without extracelluar secretion. These results indicate that the HpaA of H. pylori K51 was successfully expressed in L. lactis, and suggest that the recombinant L. lactis expressing HpaA may be applicable as an oral vaccine to induce a protective immune response against H. pylori.

Optimum Conditions for the Formation of Tetramethylpyrazine Flavor Compound by Lactococcus lactis ssp. lactis biovar. diacetilactis FC1

  • Kim, Kyoung-Heon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.285-287
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    • 1991
  • To produce the tetramethylpyrazine (TMP) flavor compound, Lactococcuss lactis subsp. lactis biovar. diacetilactis (L. diacetilactis) FC1 was cultivated in the TMP medium containing 3% (w/v) of Na-citrate and 6% (w/v) arginine-HC1 as substrates of acetoin and $NH_3$, respectively, which are the two precursors of the TMP. After 19-day fermentation at $34^{\circ}C$, 0.57 g/l or 4.19 mmole/l of the TMP was produced. This was the first result showing that the TMP could be produced by L. diacetilactis.

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Molecular Cloning of a $\beta$-D-Galactosidase Gene from Lactococcus lactis subsp. lactis 7962

  • CHANG, HAE-CHOON;YANG-DO CHOI;HYONG-JOO LEE
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.386-390
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    • 1996
  • The ${\beta}$-galactosidase gene from Lactococcus lactis subsp. lactis ATCC 7962 was cloned and its enzymatic properties were characterized, with a view to assessing its potential use as a selection marker in the food-grade cloning vector. Chromosomal DNA from L. lactis subsp. lactis 7962 was cleaved with PstI and ligated into pBR322 for transformation into Escherichia coli TGl. Transformants showing ${\beta}$-galactosidase activity possessed the pBR322 plasmid containing a 10 kilobase (kb) PstI fragment and this plasmid was named pCKL11. The cloned ${\beta}$-galactosidase gene came from the chromosomal DNA of L. lactis subsp. lactis 7962 was confirmed by Southern hybridization. A restriction map of pCKL11 was constructed from the cleavage of both pCKL11 and the purified 10kb insert fraqment. The. optimum pH of the ${\beta}$-galactosidase determined with the E. coli harboring the pCKL11 was 7.0. The optimum temperature was $50^{\circ}C$, while the pI of the enzyme was 7.4. These values were the same as those of the enzyme from the parent strain.

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Effect of peptide on the mixed fermentation of Lactobacillus helveticus YM-1 and Streptococcus lactis $ML_3$ in skim milk (Lactobacillus helveticus YM-1과 Streptococcus lactis $ML_3$의 혼합발효에 미치는 peptide의 영향)

  • 박정길;류인덕;유주현
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.487-493
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    • 1986
  • Lactobacillus helveticus YM-1and Streptococcus lactis Ml$_3$ were inoculated together in reconstituted non-fat skim milk medium, and then their proteolytic activity and stimulatory compound for acid production were investigated. Significant difference between Lactobacillus helveticus YM-1 and Streptococcus lactis Ml$_3$was observed in the proteolytic activities. The proteolytic activity of Lactobacillus helveticus YM-1 and Streptococcus lactis Ml$_3$ was 105 $\mu\textrm{g}$/$m{\ell}$ and 30 $\mu\textrm{g}$/$m{\ell}$ when converted the amounts of hydrolysates of milk protein determined by Folin Ciocaleau phenol method into their tyrosine equivalent Stimulatory compounds in cell-free filtrate of Lactobacillus helveticus YM-1were identified as peptide with a molecular weight of approximately 4, 300 for the acid production by Streptococcus lactis Ml$_3$. Some kinds of amino acids, such as histidine, lysine, arginine and glutamic acid, were rich in acid hydrolysates of peptide. Among amino acids, histidine, glutamic acid and phenylalanine stimulated acid production, on the contrary isoleucine inhibited.

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Inhibition of Listeria monocytogenes in Fresh Cheese Using a Bacteriocin-Producing Lactococcus lactis CAU2013 Strain

  • Yoon, Sung-Hee;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.42 no.6
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    • pp.1009-1019
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    • 2022
  • In recent years, biocontrol of foodborne pathogens has become a concern in the food industry, owing to safety issues. Listeria monocytogenes is one of the foodborne pathogens that causes listeriosis. The major concern in the control of L. monocytogenes is its viability as it can survive in a wide range of environments. The purpose of this study was to isolate lactic acid bacteria with antimicrobial activity, evaluate their applicability as a cheese starter, and evaluate their inhibitory effects on L. monocytogenes. Lactococcus lactis strain with antibacterial activity was isolated from raw milk. The isolated strain was a low acidifier, making it a suitable candidate as an adjunct starter culture. The commercial starter culture TCC-3 was used as a primary starter in this study. Fresh cheese was produced using TCC-3 and L. lactis CAU2013 at a laboratory scale. Growth of L. monocytogenes (5 Log CFU/g) in the cheese inoculated with it was monitored during the storage at 4℃ and 10℃ for 5 days. The count of L. monocytogenes was 1 Log unit lower in the cheese produced using the lactic acid bacteria strain compared to that in the cheese produced using the commercial starter. The use of bacteriocin-producing lactic acid bacteria as a starter culture efficiently inhibited the growth of L. monocytogenes. Therefore, L. lactis can be used as a protective adjunct starter culture for cheese production and can improve the safety of the product leading to an increase in its shelf-life.

Antifungal Activity of Lactic Acid Bacteria Isolated from Kimchi Against Aspergillus fumigatus

  • Kim, Jeong-Dong
    • Mycobiology
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    • v.33 no.4
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    • pp.210-214
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    • 2005
  • More than 120 isolates of lactic acid bacteria obtained from Kimchi was screened for antifungal activity against Aspergillus fumigatus. Approximately 10% of the isolates showed inhibitory activity and only 4.16% (five isolates) exhibited strong activity against the indicator fungus A. fumigatus. The five isolates showed a wide rang of antifungal activity against A. flavus, Fusarium moniliforme, Penicillium commune, and Rhizopus oryzae. They were identified by 16S rDNA sequencing as Lactobacillus cruvatus, L. lactis subsp. lactis, L. casei, L. pentosus, and L. sakei. The effect of Lactobacillus on mycelial growth and fungal biomass as well as its ability to produce toxic compounds were determined. The results indicate that the three species, Lactobacillus casei, L. lactis subsp. lactis, and L. pentosus, are active against A. fumigatus.

Characterization of Bacteriocin Production by Lactococcus lactis LAB3113 Isolated from Kimchi

  • Shin, Jong-Yeun;Cheol Ahn
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.101-108
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    • 1997
  • A lactic acid bacterium LAB3113, isolated from traditionally fermented Kimchi was found to produce bacteriocin whose activity was very specific toward lactobacilli and not effective against any other bacteria. Lactobacilli affected by the inhibitory substance included Lactobacillus delbrueckii-lactis, L. johnsonii, L. gsseri, and L. curvatus. Based upon biochemical and physiological characteristics, LAB3113 was classified as Lactococcus lactis, and its bacteriocin was named as lactococcin K3113. Lactococcus lactis. LAB3113 produced bacteriocin at th early stage of growth and the concentration of the bacteriocin did not decrease even after alt stationalry phase. Optimal temperature of bacteriocin production was $25^{\circ}C$ at the initial pH 7.0. Partially purified lactococcin K3113 was completely inactivated by protease, but not affected by lipase, lysozyme and RNase. The bacteriocin was very heat-stable even after autoclaving for 20 min. It was also stable in pH changes, an was not affected by th presence of solvents. lacotococcin K3113 appeared to act in bactericidal mode against L. delbrueckii-lactis ATCC4797. Molecular weight of lactococcin K3113 was calibrated as 10,500 dal by SDS-PAGE an activity staining. Lactococcus lactis LAB3113 had four residential plasmids of 3.7kb, 11.2kb, 15.5kb, and 48kh in molecular sizes. Plasmid profile analysis of mutant strain revealed that 15.5 kb plasmid was re-sponsible for the production of lactococcin K3113 and its immunity to the bacteriocin.

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Studies on Antioxidative Effect and Lactic Acid Bacteria Growth of Persimmon Leaf Extracts (감잎 추출물의 유산균 생육 및 항산화 효과에 관한 연구)

  • Woo Jun-Young;Paek Nam-Soo;Kim Young-Man
    • The Korean Journal of Food And Nutrition
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    • v.18 no.1
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    • pp.28-38
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    • 2005
  • This study was investigated the growth effect of persimmon leaf extracts on various Lactic acid bacteria, the effect as freeze-dry cryoprotectan, by the changes of vitamin C content, electron donating ability, total cell count and SOD-like activity. The total cell count of L. acidophilus MG501, L. brevis MG19, L. bulgaricus MG515, L. lactis MG530 and L. casei MG311 in the absence of persimmon leaf extracts(10%) at 37℃ after 48hr were 3.2×10/sup 9/cfu/mL, 2.9×10/sup 9/cfu/mL, 1.1×10/sup 9/cfu/mL, 1.6×10/sup 9/cfu/mL and 3×10/sup 9/cfu/mL, respectively, and pH and total acidity of those were 3.82∼3.88, and 1.697∼1.842. On the other hand, the total cell count of L. acidophilus MG501, L. brevis MG19, L. bulgaricus MG515, L. lactis MG530 and L. casei MG311 in the presence of persimmon leaf extracts at 37℃ after 48hr were 4.4×10/sup 9/cfu/mL, 4.3×10/sup 9/cfu/mL, 2×10/sup 9/cfu/mL, 3.3×10/sup 9/cfu/mL, and 3.4×10/sup 9/cfu/mL, respectively, and pH and total acidity of those were 3.74∼3.82 and 1.528∼1.805. The total cell counter of lactic acid bacteria in the presence of persimmon leaf extracts(10%) at 37℃ after 48hr higher than those in the absence of persimmon leaf extracts. In freeze-dry chryoprotectan, the survival rate of L. acidphilus MG501 decreased about 30%. In case of L. brevis MG19 and L. bulgaricus MG515, the survival rate decreased about 10%. However, the survival rate of L. lactis MG530 increased about 10% and L. casei MG311 showed the almost same effect. The changes of vitamin C content were the highest in L. lactis MG530(190.26mg/mL) and the lowest in L. acidophilus MG501(56.05 mg/mL). The electron donating ability indicated to L. acidophilus MG501, L. brevis MG19, L. bulgaricus MG515, L. lactis MG530 and L. casei MG311 added by 10% persimmon leaf extracts, respectively. when 10% persimmon leaf extracts were added L. brevis MG19, the electron donating ability is the highest of all lactic acid bacteria. The SOD-like activity of L. acidophilus MG501, L. brevis MG19, L. bulgaricus MG515, L. lactis MG530 and L. casei MG311 showed lower than that of control.

Application of Lactococcus lactis HY7803 into Soybean Fermentation for Production of Glutamic Acid (글루탐산 생산을 위한 Lactococcus lactis HY7803 균주의 대두 발효 적용)

  • Jungmin Lee;Sojeong Heo;Jihoon Choi;Eunji Pyo;Myounghee Lee;Sangick Shin;Jaehwan Lee;Junglyoul Lee;Do-Won Jeong
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.53-58
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    • 2023
  • The glutamic acid producing strain for development savory taste enhancing foods was identified the possibility through application into soybean fermentation. To check the effects on glutamic acid production during soybean fermentation, Lactococcus lactis HY7803 was introduced as a starter. The soybean samples were analyzed on days 0, 7, 14 and 21. The numbers of bacteria decreased gradually, while the content of amino-type nitrogen increased during fermentation in the soybean with L. lactis HY7803. Glutamic acid content in soybeans with L. lactis HY7803 increased from 114.99 ± 9.37 pmol/ul on day 0 to 138.14 ± 1.76 pmol/ul on day 21, showing an overall higher amino acid content than soybeans without L. lactis HY7803 and similar content to soybeans with Aspergillus oryzae SNU-G. It was clearly distinguished through principal component analysis. Consequently, our results indicate that L. lactis HY7803 is available as a fungus replacement and may be a good starter strain for enhancing savory taste in vitro as well as soybean fermentation.