• Journal of Dairy Science and Biotechnology
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• v.35 no.3
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• pp.196-201
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• 2017

Recent studies have reported that certain lactic acid bacteria and their metabolites, such as exopolysaccharides (EPSs), have immunological effects and can modulate the immune system following oral administration. Lactococcus lactis subsp. cremoris FC is a lactic acid bacteria isolated from fermented milk from Caucasians and has been shown to produce EPSs. In this study, the effects of lactoferrins (apo-lactoferrin, holo-lactoferrin, and native-lactoferrin) and transferrins (apo-transferrin and holo-lactoferrin) on the growth of L. lactis subsp. cremoris FC were examined. The addition of lactoferrins and transferrins to L. lactis subsp. cremoris FC cultures was found to be effective at concentrations of 0.5 or 1 mg/mL.

• Journal of Dairy Science and Biotechnology
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• v.37 no.2
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• pp.129-135
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• 2019

This study investigated the effects of heat-treated and non-heat-treated bovine lactoferrin on the growth of Lactococcus lactis subsp. cremoris JCM 20076. The addition of heat-treated and non-heat-treated bovine lactoferrin in adjusted MRS medium stimulated the growth of Lc. cremoris JCM 20076. Heat-treated bovine lactoferrin had a greater impact on the growth of Lc. cremoris JCM 20076 compared to that with non-heat-treated bovine lactoferrin. Bovine lactoferrin heated at $65^{\circ}C$ for 30 min stimulated the growth of the bacteria more than that heated at $80^{\circ}C$ for 5 min. Furthermore, the growth of Lc. cremoris JCM 20076 increased substantially with heat-treated bovine lactoferrin at a concentration of 1 mg/mL.

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.233-239
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• 1994

The characteristics of the bacteriophage resistant Lactococcus lactis subsp. cremoris ATCC 11602-A1, the phage-resistant mutant of Lactococcus lactis subsp. cremoris ATCC 11602, was examined. Electron microscopic study of phage adsorption to A1 revealed that after 10 min. incubation of the host-phage mixture, A1 did not show phage adsorption, and after 60 min. did not show a real burst and the release of new phage particles which could be detected in the mixture of its parent strain and phage. However, the phage adsorption rate of A1 after SDS treatment increased to 98%. Moreover, when the cell walls from A1 and parent strain, and the polysaccharide(PS) and peptidoglycan(PG) of their cell wall were mixed with phage and incubated for 15 min., PS and PG from A1 did not bind phage, but only SD-treated cell wall bound phage, and the cell wall and PS of parent strain bound phage. Both A1 and parent strain treated with 0.2 N HCl-and 5% TCA(100$$C) did not bind phage. The results suggest that the phage receptor is still present in the cell wall of the A1, but a cell wall constituent hydrolyzed by SDS blocks phage adsorption by masking the phage receptor. It also suggests that the phage receptor of parent strain is associated with PS of the cell wall.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.293-298
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• 1993

The ppage resistance mechanism of Lactococcus lactis subsp. cremoris ATCC 11602-A1 was investigated. When parent and A1 were incubated at 30 and 40$^{\circ}C$, A1 grew well and multiplication of phage(MOI=1)on A1 slightly occurred at 40$^{\circ}C$ in contrast with parent. There was a great difference of proteolytic activity between parent and A1, irrespective of the temperature. As a result of ADS treatment oon culture broth, survival rate of A1 was 27% at the lethal concentration of parent and adsorption rate of phage was increased to 95~97%, which was considered to come from the exposure of phage receptor site masked by an unknown component. These results suggest that acridine orange (AO) treatment leads to the modification of cell wall, conferring resistance to high temperature and lytic phage. No change in plasmid profiles of A1 at 30 and 40$^{\circ}C$ were found, which suggests that plasmid is not relative to temperature-resistance of A1.

• Journal of Microbiology and Biotechnology
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• v.10 no.4
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• pp.539-543
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• 2000

A new bacteriocin, named lacticin 10790, was purified from Lactococcus lactis subsp. cremoris KFCC 10790 by sequential adsorption, immobilized metal-affinity, cation-exchange, and $C_{18}$ reverse-phase chromatographies. The molecular mass of the bacteriocin was estimated to be between 3,000 and 3,500 Da. Lacticin 10790 showed a broad antimicrobial spectrum against many gram-positive bacteria. The bacteriocin was stable to heat and in the pH range between 2 and 6. Lacticin 10790 was destroyed by digestion with proteases and exhibited a bactericidal mode of action. An amino acid composition analysis of purified lacticin 10790 revealed a high concentration of hydrophobic amino acids. The N terminus of the bacteriocin was found to be blocked, upon analysis by Edman degradation. The results suggest that lacticin 10790 is a class I bacteriocin.

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.91-101
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• 2018

The aim of this study was to transfer the 18.5 kb gene clusters coding for 17 genes from Lactobacillus rhamnosus to Lactococcus lactis subsp. cremoris MG1363 in order to determine the effect of host on exopolysaccharide (EPS) production and to provide a model for studying the phosphorylation of proteins which are proposed to be involved in EPS polymerization. Lactobacillus rhamnosus RW-9595M and ATCC 9595 have 99% identical operons coding for EPS biosynthesis, produced different amounts of EPS (543 vs 108 mg/l). L. lactis subsp. cremoris MG1363 transformed with the operons from RW-9595M and ATCC 9595 respectively, produced 326 and 302 mg/l EPS in M17 containing 0.5% glucose. The tyrosine protein kinase transmembrane modulator (Wzd) was proposed to participate in regulating chain elongation of EPS polymers by interacting with the tyrosine protein kinase Wze. While Wzd was found in phosphorylated form in the presence of the phosphorylated kinase (Wze), no phosphorylated proteins were detected when all nine tyrosines of Wzd were mutated to phenylalanine. Lactococcus lactis subsp. cremoris could produce higher amounts of EPS than other EPS-producing lactococci when expressing genes from L. rhamnosus. Phosphorylated Wzd was essential for the phosphorylation of Wze when expressed in vivo.

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.240-245
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• 1994

Relation the phage defense mechanism of phage resistant Lactococcus lactis subsp. cremoris ATCC 11602-A1 to its cell wall components was investigated. To determine whether teichoic acid which is known to be one of the phage receptor site present on the cell wall, phage adsorption was examined after treatment 5% TCA(60%$\CIRC $C) and concanavalin A to the cell wall of A1 and parent strain. However, the adsorption rate of two strains did not change. Total amount of phosphate after TCA treatment did not change in both strains, but a difference between the two strains was observed. Ribitol and glycerol, components of teichoic acid, could not be detected in the cell walls of two strains by GC analysis. These results suggest that although teichoic acid was not present in the cell walls of both strains, the composition of cell wall of two strains was not identical. Measurement of amount of protein and SDS-polyacryamide gel electrophoresis were carried out to examine the involvement of cell wall protein in phage resistance, showing that protein is nothing to do with phage adsorption of parent strain, but phage resistance of A1 is related to protein. Cell wall carbohydrates of A1 contained rhamnose, glucose, and galactose. Total amount of carbohydrate of 1% SDS-treated A1 cell wall was reduced to the level of parent strain. The results suggest that phage resistance of A1 was due to the presence of a higher level of carbohydrates then parent strain, and to interaction of carbohydrate and protein.

• Journal of Life Science
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• v.6 no.1
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• pp.27-33
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• 1996

Reactions taken place by a mixed culture of Lactococcus lactis subsp. cremoris KH (lac$^{+}$ prt$^{+}$ ) and KHA (lac$^{-}$ prt$^{-}$ ) and KHA (lac prt ) in cheese ripening have been investigated. Growth characteristics of the mixed culture showed commensalism, and the amounts of proteinases of the mixed culture were small enough. From these results, it is concluded that the production of bitter taste by the mixed culture is a small matter, even if the density of the mixed culture is highly maintained during cheese ripening. Hence, the mixed culture of KH and KHA cells could be a good cheese starter in accelerating the process of cheese ripening.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.1052-1056
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• 2004

A secretion signal sequence-selection vector (pGS40) was constructed based on an $\alpha$-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore $\alpha$-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgamo (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.

• Journal of Microbiology and Biotechnology
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• v.30 no.9
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• pp.1404-1411
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• 2020

Lactic acid bacteria (LAB) play an important role in dairy fermentations, notably as cheese starter cultures. During the cheese production and ripening period, various enzymes from milk, rennet, starter cultures, and non-starter LABs are involved in flavor formation pathways, including glycolysis, proteolysis, and lipolysis. Among these three pathways, starter LABs are particularly related to amino acid degradation, presumably as the origins of major flavor compounds. Therefore, we used several enzymes as major criteria for the selection of starter bacteria with flavor-forming ability. Lactococcus lactis subsp. lactis LDTM6802 and Lactococcus lactis subsp. cremoris LDTM6803, isolated from Korean raw milk and cucumber kimchi, were confirmed by using multiplex PCR and characterized as starter bacteria. The combinations of starter bacteria were validated in a miniature Gouda-type cheese model. The flavor compounds of the tested miniature cheeses were analyzed and profiled by using an electronic nose. Compared to commercial industrial cheese starters, selected starter bacteria showed lower pH, and more variety in their flavor profile. These results demonstrated that LDTM6802 and LDTM6803 as starter bacteria have potent starter properties with a characteristic flavor-forming ability in cheese.