• Journal of Microbiology and Biotechnology
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• v.2 no.2
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• pp.73-77
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• 1992

The effect of a nisin-producing Lactococcus lactis spp. lactis (L. lactis) on the growth and attachment of Listeria monocytogenes Scott A and Brie 1 on stainless steel and their growth in Brain Heart Infusion broth was determined. Viable cells of Listeria decreased rapidly after 9~12 hr of incubation at $21^{\circ}C$ and after 6~9 hr of incubation at $32^{\circ}C$ in the presence of L. lactis. The number of L. monocytogenes Scott A attached to stainless steel in pure culture was $2.5{\times}10^3/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}2.3{\times}10^3/\textrm{cm}^2{\;}at{\;}32^{\circ}C$ after 48 hr of incubation, but was only $10/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}1.1{\times}10/\textrm{cm}^2{\;}at{\;}32^{\circ}C$ in the presence of L. lactis. Results from L. monocytogenes strain Brie 1 were similar to those from strain Scott A. The population of L. monocytogenes Scott A which attached to stainless steel with previously adherent L. lactis was $1.8{\times}10^2/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}8.2{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$, whereas the population attached to sterile stainless steel was $1.2{\times}10^3/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}2.1{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$. For L. monocytogenes Brie 1, the attached population of the control was $1.6{\times}10^4/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}3.2{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$, and on stainless steel with adherent L. lactis, it was $1.1{\times}10/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}6.9{\times}10/\textrm{cm}^2{\;}at{\;}32^{\circ}C$. Surface adherent L. lactis was less inhibitory to attachment of L. monocytogenes on stainless steel than a liquid culture inoculum. Listeria attached to stainless steel survived dry storage for 20 days both in the presence and absence of adherent lactococci.

• Journal of Microbiology and Biotechnology
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• v.3 no.4
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• pp.217-223
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• 1993

To investigate the nature and location of the nisin-resistance determinant of Lactococcus lactis subsp. lactis 7962 (L. lactis 7962), a total plasmid DNA prepared from L. lactis 7962, a nisin producer, was used to transform L. lactis subsp. lactis LM0230, a plasmid-free and nisin-sensitive strain, by protoplast mediated transformation procedures. All of the nisin-resistant transformants acquired the ability to utilize sucrose at the same time, confirming the close linkage between these two determinants in L. lactis 7962. The plasmid DNA profiles of a few selected nisin-resistant transformants were examined by agarose gel electrophoresis. No common plasmid was found among the transformants and some small plasmids previously not present in L. lactis 7962 were detected. These transformants were named as L. lactis KL1, KL2, KL3, KL4, or KL5, respectively based on their plasmid profiles. Growth curves of all transformants were similar to that of L. lactis LM0230, but different from that of L. lactis 7962. L. lactis KL5 showed the highest level of resistance to nisin, growing up to 1, 200 IU nisin/ml after 40 hr incubation. Some nisin-sensitive derivatives of KL1 or KL2 were obtained by plasmid curing experiments. The plasmid DNA profiles of the nisin-sensitive KL1 derivatives were apparently the same as that of the KL1. All of the nisin-sensitive KL2 derivatives were plasmid-free, but a nisin-resistant strain with no apparent plasmid was also obtained. These results indicate that the nisin-resistance of the $Nis^r$ transformants is presumably mediated by the chromosomally located gene(s) rather than plasmid-encoded gene(s).

• Microbiology and Biotechnology Letters
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• v.19 no.6
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• pp.619-623
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• 1991

To investigate nisin production and resistance of Lactococcus lactis ssp. tactis ML (L. lactis $ML_8$, effects of medium, pH of culture broth, and cell growth on the nisin activity, and effect of nisin with or without $Ca^[2+}$ ion on the growth of L. lactzs were analyzed. In the bio-assay of nisin by the agar diffusion method, inhibition-zone diameter of Micrococcus Javus was propotional to the logarithm of nisin concentration ranged 0.5~20 unitlml (12.5~500 ng/mf). Nisin activity of the pasteurized culture filtrates of L. lactis MLs was high at pH 2!3 but was inactivated completely at pH over 6.0. Nisin production of the L. lactis $ML_8$ cultured on LTB broth increased at late logarithmic phase and reached 10.5 unitlml after 16 hr. The cell growth of L. lactis LM 0230, a plasmid free and nisin sensitive strain, was inhibited on agar medium containing 7 unitlrnl of nisin, while L. lactis $ML_8$ showed high survival ability at 20 unitld of nisin. When 40 mM $Ca^[2+}$ ion was added to Elliker broth with 8 unitlml of nisin, the growth pattern of L. lactis $ML_8$ was similiar to that on control medium which did not contain nisin and $Ca^[2+}$ ion, and this suggested that $Ca^[2+}$ increased the nisin resistance of the L. lactis.

• Journal of Pharmaceutical Investigation
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• v.35 no.2
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• pp.75-80
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• 2005

A promising application of Lactococcus lactis is its use as live vehicles for production and delivery of heterologous proteins of vaccines and therapeutic substances. Because L. lactis has GRAS ('generally regarded as safe') status, we tested whether L. lactis could function as the carrier of the Ll protein of human papillomavirus (HPV) type 16. The RNA level expression of Ll gene was detected in L. Lactis. The Ll protein was expressed in L. lactis with Ll gene. The growth of strains L. lactis with an empty plasmid (pAMJ328) and L. lactis with Ll-encoding plasmid (pAMJ328-Ll) was slightly decreased in comparison with the growth of strains L. lactis (wild type). However, all the three strains of L. lactis maintained the ability to ferment sugars primarily into lactic acid, indicating that Ll protein did not affect the biochemical property of L. lactis. These results suggest that L. lactis, capable of carrying Ll protein, might be further developed as a biocompatible oral protein delivery system.

• Microbiology and Biotechnology Letters
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• v.33 no.4
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• pp.241-247
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• 2005

A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a $\beta$-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.732-736
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• 1999

The superoxide dismutase (SOD) in Lactococcus lactis was measured quantitatively and qualitatively under various culture conditions. The L. lactis SOD was induced by oxidative stress. As the concentration of paraquat to produce superoxide radicals increased, the growth of L. lactis decreased with concomitant increase of SOD activity. The SOD activity was found to be growth-phase dependent: when aerobically grown cells entered to the stationary phase, the activity increased gradually until the late stationary phase. From inhibition studies, L. lactis SOD was found to be insensitive to KCN and $H_2O_2$ which are known to inhibit Cu/ZnSOD and FeSOD, respectively. Moreover, as the concentration of manganese in the medium increased, the activity of SOD also increased. These data strongly suggested that L. lactis possessed a single manganese-containing SOD (MnSOD). Finally, a putative sod gene fragment of 510 bp was identified in L. lactis using a polymerase chain reaction (PCR) with degenerate primers designed from the deduced DNA sequences of known SOD genes.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.4
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• pp.549-557
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• 2000

Dental caries is a bacterial disease of the dental hard tisssus, characterized by a localized, progressive, molecular disintegration of tooth structure. The action of Leuconostoc lactis 51 about plaque formation and replication by Streptococcus mutans was studied as follows. 1. Lower amount of plaque was produced at the mixed culture of S. mutans and L. lactis 51 than S. mutans alone on the wires in the beaker. 2. Fewer cells of S. mutans were replicated at the mixed culture of S. mutans and L. lactis 51 than S. mutans alone. 3. In M17Y broth, viable cells of S. mutans and L. lactis 51 increased for 12 hours, and decreased for 24 hours. In M17YS broth, viable cells of S. mutans showed time-dependent decrease at mixed culture of S. mutans and L. lactis 51. 4. The culture supernatant of L. lactis 51 didn't inhibit the replication of S. mutans and the formation of artificial plaque. 5. Sucrose and frutose were extracted from the culture supernatant of L. lactis 51 in M17YS broth. These results suggest that L. lactis 51 isolated from the oral cavity inhibits the replication of S. mutans and the formation of artificial plaque.

• Journal of Microbiology and Biotechnology
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• v.33 no.6
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• pp.823-830
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• 2023

Lactococcus lactis is a lactic acid bacterium and used in the dairy food industry. The ameliorating effects of Lactobacillus species on atopic dermatitis (AD) have been extensively studied, but the specific effect of L. lactis strains has not yet been investigated. In this study, the efficacy of L. lactis LB 1022, isolated from natural cheese, was evaluated using RAW 264.7, HMC-1 and HaCaT cell lines and an ovalbumin-sensitized AD mouse model. L. lactis LB 1022 exhibited nitric oxide suppression and anti-allergy and anti-inflammatory activity in vitro. Oral administration of L. lactis LB 1022 to AD mice significantly reduced the levels of IgE, mast cells, and eosinophils, and a range of T cell-mediated T helper Th1, Th2, and Th17-type cytokines under interleukin (IL)-10, transforming growth factor-β (TGF-β), thymus and activation-regulated chemokine (TARC), and thymic stromal lymphopoietin (TSLP). In addition, L. lactis LB 1022 treatment increased the concentration of short-chain fatty acids. Overall, L. lactis LB 1022 significantly modulated AD-like symptoms by altering metabolites and the immune response, illustrating its potential as candidate for use in functional food supplements to alleviate AD.

• Food Science and Preservation
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• v.6 no.4
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• pp.442-447
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• 1999

A curd yogurt was prepared by fermenting milk added with skim milk powder and purple sweet potato by culture of 5 types of lactic acid bacteria(Lactobacillus delbruekii sub. sp. lactis, Streptococcus lactis, acidity, number of viable cell, stability of purple sweet potato's pigment and keeping qualify. Among the organisms tested, the acid production and number of viable cell by the culture of L bulgaricus remarkably increased for the first 12 hem which showed 1.04${\times}$10$\^$9/ CFU/mL in number of viable cell and 4.22 In pH where as fermentation by the culture of B. bifidum was slow. After 36 hours of incubation which showed 3.3 ${\times}$ l0$\^$8/ CFU/mL in number of viable cell and 5.1 in pH. In stabilities of purple sweet potato anthocyanin pigment n fermentation, yogurt by B. bifidum was found to be most stable followed by Leuc. lactis, L. delbruekii sub. sp. lactis, L bulgaricus, but yogurt by St. lactis was not stable. When curd yogurt added with Purple sweet Potato was kept at 2∼3$^{\circ}C$ for 14 day, its keeping quality(pH, titratable acidity, number of viable cell) was relative good except product by L. bulgaricus was found to be decreased most of viable cell. After 2 weeks of keeping, pigment of yogurt was decreased by B. bifidum, stable by L. delbruekii sub. sp. lactis.

• Journal of environmental and Sanitary engineering
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• v.12 no.1
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• pp.85-95
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• 1997

Behavior of Listeria monocytogenes in Skim milk during fermentation by Lactobacillus bulgaricus YI-2 and Streptococcus lactis FYI-1 were determined. Autoclaved skim milk was inoculated with ca. 10$^{3}$ L. monocytogenes (Strain LM91-1 or LM 96-2) cells/ml, and with 5.0, 1.0, 0.5 or 0.1% of a milk culture of either L. bulgaricus TI-2 or S. lactis FYI-1. Skim milk containing ca. 10$^{3}$ L. monocytogenes was incubated at 37 or 42$\circ $C for 15 h with L. bulgaricus YI-2, and at 21 or 30$\circ $C for 15 h with S. lactis FYI-1. Cultured skim milks were stored at 4$\circ $C in the refrigerater. Samples were plated on Oxford Agar with oxford antimicrobic supplement to enumerate L. monocytogenes and on either modified MRS agar to enumerate lactic acid bacteria. L. monocytogenes survived the 15-h fermentation with S. lactis FYI-1 in all combinations of level of inoculum and temperature of incubation, but inhibition of growth ranged from 94 to 100%. When incubated with over the 1.0% of L. bulgaricus, L. monocytogenes inhibited or disappeared in fermented skim milk from 9 h after incubation. Especially, incubation at 42$\circ $C with 5.0% L. bulgaricus YI-2 as inoculum appeared to be the most effective inhibitory combination for strain LM 91-1, causing 100% inhibition in growth based on maximum papulation attained. In most instances of incubated with L. bulgaricus YI-2, growth of the pathogene appeared to be completely inhibited when the pH dropped below 4.38.