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Immunostimulatory Effect of Heat-Killed Probiotics on RAW264.7 Macrophages
( Hye-ji Noh ),( Jung Min Park ),( Yoo Jin Kwon ),( Kyunghwan Kim ),( Sung Yurb Park ),( Insu Kim ),( Jong Hyun Lim ),( Byoung Kook Kim ),( Byung-yong Kim ) 한국미생물생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.5
Probiotics modulate the gut microbiota, which in turn regulate immune responses to maintain balanced immune homeostasis in the host. However, it is unclear how probiotic bacteria regulate immune responses. In this study we investigated the immunomodulatory effects of heat-killed probiotics, including Lactiplantibacillus plantarum KC3 (LP3), Lactiplantibacillus plantarum CKDB008 (LP8), and Limosilactobacillus fermentum SRK414 (LF4), via phagocytosis, nitric oxide (NO), and pro-inflammatory cytokine production in macrophages. We thus found that heat-killed LP8 could promote the clearance of foreign pathogens by enhancing the phagocytosis of macrophages. Treatment with heat-killed LP8 induced the production of NO and pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β. In addition, heat-killed LP8 suppressed the production of NO and cytokines in LPS-induced RAW264.7 cells, suggesting that heat-killed LP8 exerts immunomodulatory effects depending on the host condition. In sum, these results indicate that heat-killed LP8 possesses the potential for immune modulation while providing a molecular basis for the development of functional probiotics prepared from inactivated bacterial cells.
김병용,Bonggyu Min,Yoo-Jin Kwon,SUNG-YURB PARK,Jong Hyun Lim,Chang-Hun Shin,Byoung Kook Kim 건강기능식품미래포럼 2022 건강기능식품미래포럼 학술지 Vol.2 No.4
Lactiplantibacillus plantarum CKDB008 was isolated from a Korean fermented food, kimchi. In order to investigate its potential as a probiotic in the food industry, the genome sequence of L. plantarum CKDB008 was analyzed to confirm its probiotic properties and safety at the genome level. The CKDB008 strain has one circular chromosome of 3.2 Mbp, with 44.6% guanine-cytosine content. A comparative genomic analysis confirmed that its genome was similar to those of 19 L. plantarum strains used as probiotics. Pan-genome analysis of 20 L. plantarum strains identified 6,236 genes, including 2,006 core, 255 soft core, 1,485 shells, and 2,490 cloud genes. These results reveal the ‘open’ nature of L. plantarum pan-genome. Among the predicted 3,129 protein coding genes, the genome contains genes associated with important characteristics of probiotics such as protease system, amino acid biosynthesis, major carbohydrate-active enzymes, stress resistance, and cell adhesion. In addition, genes encoding bacteriocins, immunoregulatory, and gamma-aminobutyric acid biosynthesis were also identified, but safety-related genes related to antibiotic resistance and virulence factors were not. The results obtained strongly suggest that CKDB008 strain is genetically safe and has the desired properties required for a probiotic as well as support that L. plantarum CKDB008 strain has a potential as a probiotic that can be used for the industrial purpose.
Streptococcus thermophilus KCTC14471BP의 안전성 평가 및 실크 피브로인 코팅의 면역 증진 효능
권기철,Heo Bohye,손성호,Kwon Yoo Jin,권미진,장주현,신혜정,Kim Insu,Park Sung Yurb,김경환,Kim Byoung Kook,신창훈,김병용 한국유산균프로바이오틱스학회 2022 Current Topic in Lactic Acid Bacteria and Probioti Vol.8 No.1
Probiotic strain is known to regulate the immune system by colonizing in the intestine and interact with intestinal cell receptors of lymphoid tissue. In this study, safety of Streptococcus thermophilus KCTC14471BP and silk fibroin coating effects was evaluated with respect to mucin binding abilities and immune system modulation. S. thermophilus KCTC14471BP was coated with silk fibroin by adding 1% water-soluble calcium and 0.1% silk fibroin. S. thermophilus KCTC14471BP showed the high activities of leucine arylamidase and β-galactosidase. Regarding the antibiotic resistance tests, S. thermophilus KCTC14471BP was susceptible to ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline, and chloramphenicol. S. thermophilus KCTC14471BP coated with silk fibroin showed the improved mucin binding ability from 16.1% to 71.3% and was confirmed to have no cytotoxicity against RAW 264.7 macrophage. S. thermophilus KCTC14471BP coated with silk fibroin showed dose-dependently significant increases in pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. These results suggested that S. thermophilus KCTC14471BP can be expected as a promising probiotic bacteria for applications to food industries such as fermentation or functional foods.
Kim Myung-Soo,Min Bonggyu,Kim Insu,Kwon Yoo Jin,Park Sung Yurb,Kim Byung-Yong,Kim Byoung Kook,박성선 건강기능식품미래포럼 2021 건강기능식품미래포럼 학술지 Vol.1 No.3
Streptococcus thermophilus is a bacterial species widely used in the food industry for producing dairy fermented foods as well as probiotics. In this study, the whole genome of the S. thermophilus CKDB027 strain used in probiotic products was sequenced, and the genetic features related to its safety and functionality were determined. The CKDB027 strain has a single circular chromosome of approximately 1.88 Mb with 39.0% GC content. The genome study has shown the absence of antibiotic resistance and virulence related genes. To identify the unique genetic features of CKDB027, genomic comparisons with other 18 S. thermophilus used in probiotic industry were conducted. The results showed that the CKDB027 strain shared a common ancestor with several strains isolated from milk or yogurt. The CKDB027 strain possesses genes related to lactose catabolism, proteolysis, stress resistance, defense system, and adherence that are major function of probiotics. In addition, a gene cluster producing exopolysaccharides was detected. These findings indicate that the S. thermophilus CKDB027 strain is genetically safe and has beneficial genetic features for human health when it is used in dairy industry as a health functional food.
Safety Assessment of Lactiplantibacillus (formerly Lactobacillus) plantarum Q180
( Yoo Jin Kwon ),( Byung Hee Chun ),( Hye Su Jung ),( Jaeryang Chu ),( Hyunchae Joung ),( Sung Yurb Park ),( Byoung Kook Kim ),( Che Ok Jeon ) 한국미생물 · 생명공학회 2021 Journal of microbiology and biotechnology Vol.31 No.10
The safety of the probiotic strain Q180, which exerts postprandial lipid-lowering effects, was bioinformatically and phenotypically evaluated. The genome of strain Q180 was completely sequenced, and single circular chromosome of 3,197,263 bp without any plasmid was generated. Phylogenetic and related analyses using16S rRNA gene and whole-genome sequences revealed that strain Q180 is a member of Lactiplantibacillus (Lp., formerly Lactobacillus) plantarum. Antimicrobial resistance (AMR) genes were bioinformatically analyzed using all Lp. plantarum genomes available in GenBank, which showed that AMR genes are present differently depending on Lp. plantarum strains. Bioinformatic analysis demonstrated that some mobile genetic elements such as prophages and insertion sequences were identified in the genome of strain Q180, but because they did not contain harmful genes such as AMR genes and virulence factor (VF)- and toxin-related genes, it was suggested that there is no transferability of harmful genes. The minimum inhibition concentrations of seven tested antibiotics suggested by the European Food Safety Authority guidelines were slightly lower than or equal to the microbiological cut-off values for Lp. plantarum. Strain Q180 did not show hemolytic and gelatinase activities and biogenic amine-producing ability. Taken together, this study demonstrated the safety of strain Q180 in terms of absence of AMR genes and VF- and toxin-related genes as a probiotic strain.