• 한국식품과학회지
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• 제54권2호
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• pp.203-208
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• 2022

본 연구에서는 글루텐 분해능을 가지는 유산균을 선별하기 위해 전통식품인 젓갈을 이용하였다. 선별된 균주는 API, 16S rRNA sequencing과 현미경을 통해 간균의 형태를 갖는 Lactobacillus paracasei로 동정되었고 이를 GLU70으로 명명하기로 하였다. MRS 배지에서 GLU70의 생육특성을 조사한 결과, 6시간부터 12시간까지 대수기이며 12시간 이후부터 정지기임을 확인하였다. 이렇게 동정된 GLU70을 글루텐이 함유된 MRS배지에서 시간별로 배양하였을 때 대수기인 12시간 이후부터 24시간 이전까지 글루텐 분해가 가속화되었고, 48시간에서 최대 45% 정도의 분해율을 나타내었다. 또한 GLU70이 유산균으로서 적합하기 위해서 가져야하는 내산성, 내담즙성, 내열성에 대한 분석을 실시하였다. 많은 연구에서 Lactobacillus 종이 pH 2.5 이상에서는 균의 생존율이 높았지만, 그 이하에서는 거의 생존하지 않는다고 보고하고 있다 (Shin 등, 1999). 이에 따라 분리한 GLU70에서도 유사하게 pH 2.0에서는 균이 사멸하였고 pH 3.0부터 약 84%의 생존율을 보였고 많은 연구들과 유사한 결과를 가지는 것을 통해 우수한 내산성을 가지는 것을 확인하였다. 내담즙성의 경우 oxgall이 0.3% 함유된 배지에서 성장할 수 있을 정도의 내성을 가져야 한다는 연구결과와 비교하였을 때 GLU70은 담즙에 대하여 매우 강한 내성을 가지고 있다고 보기는 어렵다. 또한 내열성에 대한 연구결과에서는 50℃ 이상의 온도에서는 사멸하였으나 30℃에서는 높은 생존율을 나타냈다. 또한 분리된 GLU70이 실제 밀가루 반죽에서 감소하는 글루텐 함량을 분석하였을 때 12 h 배양한 배양상등액을 넣어 25℃에서 24 h 동안 발효한 밀가루 반죽에서 가장 높은 분해력을 확인하였다. 또한 밀가루 반죽의 발효 온도에 따른 글루텐 분해율을 타 균주와 비교하여 GLU70의 우수성을 판단하고자 하였다. GRAS 인정 유산균 11종과 GLU70을 각각 밀가루 반죽에 넣은 후 25℃와 35℃에서 발효하였다. 그 결과 GLU70이 타 균주들과 비교하였을 때 모든 온도에서 반죽이 흘러내리는 등의 이상현상 없이 50% 이상의 매우 높은 글루텐 분해율을 보였다. 이를 통해 GLU70이 향후 식품 산업에 다양한 활용 가능성을 지니며 글루텐 저감화 밀가루 제조 등에 적용하기에 적합하다고 제시된다.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.405-418
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• 2022

Simotang oral liquid (SMT) is a traditional Chinese medicine (TCM) consisting of four natural plants and is used to alleviate gastrointestinal side effects after chemotherapy and functional dyspepsia (FD). However, the mechanism by which SMT helps cure these gastrointestinal diseases is still unknown. Here, we discovered that SMT could alleviate gastrointestinal side effects after chemotherapy by altering gut microbiota. C57BL/6J mice were treated with cisplatin (DDP) and SMT, and biological samples were collected. Pathological changes in the small intestine were observed, and the intestinal injury score was assessed. The expression levels of the inflammatory factors IL-1β and IL-6 and the adhesive factors Occludin and ZO-1 in mouse blood or small intestine tissue were also detected. Moreover, the gut microbiota was analyzed by high-throughput sequencing of 16S rRNA amplicons. SMT was found to effectively reduce gastrointestinal mucositis after DDP injection, which lowered inflammation and tightened the intestinal epithelial cells. Gut microbiota analysis showed that the abundance of the anti-inflammatory microbiota was downregulated and that the inflammatory microbiota was upregulated in DDP-treated mice. SMT upregulated anti-inflammatory and anticancer microbiota abundance, while the inflammatory microbiota was downregulated. An antibiotic cocktail (ABX) was also used to delete mice gut microbiota to test the importance of gut microbiota, and we found that SMT could not alleviate gastrointestinal mucositis after DDP injection, showing that gut microbiota might be an important mediator of SMT treatment. Our study provides evidence that SMT might moderate gastrointestinal mucositis after chemotherapy by altering gut microbiota.

• 한국가금학회지
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• 제49권2호
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• pp.125-137
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• 2022

This study aimed to determine the effects of dietary microalgae (Tetradesmus sp. (TO)) on intestinal immunity and microbiota of pre-starter broilers. One hundred and twenty 1-day-old birds (Ross 308) were allocated to two dietary treatment groups with six blocks in a randomized complete block design. The two experimental diets consisted of a corn-soybean meal-based basal diet and a diet with 0.5% TO powder instead of cornstarch in the basal diet. After feeding the experimental diets for ten days, all birds' body weight and feed intake were measured, and representative eight birds were selected from each treatment group. Small intestinal lamina propria cells were isolated using flow cytometry to examine the frequency of immune cells. Cecal feces were harvested for 16s rRNA gut microbiota analysis and fecal IgA levels. Here, we found that 0.5% TO supplementation increased CD3⁺CD4⁺ T cells in the small intestine, but decreased CD3⁺CD8⁺ T cells in the small intestine. Gut microbial analysis showed that TO supplementation significantly increased the alpha diversity of the gut microbiome. Taxonomic analysis showed that TO treatment increased the abundance of Firmicutes and decreased that of Bacteroidetes at the phylum level. The distribution of Enterobacteriaceae containing many harmful bacteria at the family level, was lower in the TO group. In the LEfSe analysis, the TO group had a significantly enriched abundance of Agathobaculum at the genus level. Overall, results show that Tetradesmus sp. supplementation influences intestinal T-cell immunity and induces the expansion of beneficial gut microbes in pre-starter broiler chickens.

• Animal Bioscience
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• 제35권11호
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• pp.1698-1710
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• 2022

Objective: Raw potato starch (RPS) is resistant to digestion, escapes absorption, and is metabolized by intestinal microflora in the large intestine and acts as their energy source. In this study, we compared the effect of different concentrations of RPS on the intestinal bacterial community of weaned piglets. Methods: Male weaned piglets (25-days-old, 7.03±0.49 kg) were either fed a corn/soybean-based control diet (CON, n = 6) or two treatment diets supplemented with 5% RPS (RPS5, n = 4) or 10% RPS (RPS10, n = 4) for 20 days and their fecal samples were collected. The day 0 and 20 samples were analyzed using a 16S rRNA gene sequencing technology, followed by total genomic DNA extraction, library construction, and high-throughput sequencing. After statistical analysis, five phyla and 45 genera accounting for over 0.5% of the reads in any of the three groups were further analyzed. Furthermore, short-chain fatty acids (SCFAs) in the day 20 fecal samples were analyzed using gas chromatography. Results: Significant changes were not observed in the bacterial composition at the phylum level even after 20 d post feeding (dpf); however, the abundance of Intestinimonas and Barnesiella decreased in both RPS treatment groups compared to the CON group. Consumption of 5% RPS increased the abundance of Roseburia (p<0.05) and decreased the abundance of Clostridium (p<0.01) and Mediterraneibacter (p< 0.05). In contrast, consumption of 10% RPS increased the abundance of Olsenella (p<0.05) and decreased the abundance of Campylobacter (p<0.05), Kineothrix (p<0.05), Paraprevotella (p<0.05), and Vallitalea (p<0.05). Additionally, acetate (p<0.01), butyrate (p<0.05), valerate (p = 0.01), and total SCFAs (p = 0.01) were upregulated in the RPS5 treatment group Conclusion: Feeding 5% RPS altered bacterial community composition and promoted gut health in weaned piglets. Thus, resistant starch as a feed additive may prevent diarrhea in piglets during weaning.

• 농업과학연구
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• 제49권4호
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• pp.963-971
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• 2022

Production of high-quality horses is important to make the horse industry grow. Grazing during the growing period can be an important factor affecting the production of high-quality horses. The objective of this study was to determine the effects of grazing on growing horses by analyzing their blood components and intestinal microbiota. Twelve growing horses for evaluating blood components and ten growing horses for evaluating intestinal microbiota were raised for about seven months and separated by two treatments: grazing vs. stable. Complete blood count, blood chemistry, and creatine kinase levels were analyzed as blood components and a 16s rRNA gene sequence analysis was performed to analyze intestinal microbiota. Calcium ions tended to be lower in the group with grazing treatment. Alkaline phosphatase and creatine kinase tended to be higher in the group with grazing treatment. These results indicate that grazing can provide horses with more exercise than staying in stables. At the phylum level, Firmicutes/Bacteroidetes ratios in grazing and stable groups were 4.2 and 6.5, respectively. Because various studies have reported that a. high Firmicutes/Bacteroidetes ratio indicates obesity, the method of raising horses might affect their physical ability. At the species level, rates of Clostridium butyricum in grazing and stable groups were 3.2% and 13.1%, respectively. Some strains of C. butyricum can cause several diseases such as botulism. These results indicate that grazing can positively affect growing horses in terms of blood components and intestinal microbiota. Moreover, grazing can be helpful to make growing horses healthy through proper exercise.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1213-1227
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• 2023

Fetal growth restriction (FGR) is a prevalent obstetric condition. This study aimed to investigate the role of Toll-like receptor 9 (TLR9) in regulating the inflammatory response and gut microbiota structure in FGR. An FGR animal model was established in rats, and ODN1668 and hydroxychloroquine (HCQ) were administered. Changes in gut microbiota structure were assessed using 16S rRNA sequencing, and fecal microbiota transplantation (FMT) was conducted. HTR-8/Svneo cells were treated with ODN1668 and HCQ to evaluate cell growth. Histopathological analysis was performed, and relative factor levels were measured. The results showed that FGR rats exhibited elevated levels of TLR9 and myeloid differentiating primary response gene 88 (MyD88). In vitro experiments demonstrated that TLR9 inhibited trophoblast cell proliferation and invasion. TLR9 upregulated lipopolysaccharide (LPS), LPS-binding protein (LBP), interleukin (IL)-1β and tumor necrosis factor (TNF)-α while downregulating IL-10. TLR9 activated the TARF3-TBK1-IRF3 signaling pathway. In vivo experiments showed HCQ reduced inflammation in FGR rats, and the relative cytokine expression followed a similar trend to that observed in vitro. TLR9 stimulated neutrophil activation. HCQ in FGR rats resulted in changes in the abundance of Eubacterium_coprostanoligenes_group at the family level and the abundance of Eubacterium_coprostanoligenes_group and Bacteroides at the genus level. TLR9 and associated inflammatory factors were correlated with Bacteroides, Prevotella, Streptococcus, and Prevotellaceae_Ga6A1_group. FMT from FGR rats interfered with the therapeutic effects of HCQ. In conclusion, our findings suggest that TLR9 regulates the inflammatory response and gut microbiota structure in FGR, providing new insights into the pathogenesis of FGR and suggesting potential therapeutic interventions.

• Journal of Microbiology and Biotechnology
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• 제33권10호
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• pp.1292-1298
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• 2023

PAMB 00755^T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755^T was most closely related to Sphingomonas chungangi MAH-6^T (97.7%) and Sphingomonas polyaromaticivorans B2-7^T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755^T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C_18:1ω7c and/or C_18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755^T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755^T as the type strain (= KCTC 92781^T = GDMCC 1.3779^T).

• 한국응용과학기술학회지
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• 제40권5호
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• pp.1126-1135
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• 2023

인삼부산물 및 인삼밭 토양에서 분리한 9균주 중 𝛽-Glucosidase 생성 균주 GYP-1과 GYP-3-3 균주를 선발하였다. 선발된 𝛽-Glucosidase 생성 균주에 대하여 16S rRNA 유전자 염기서열과 ITS 염기서열을 기반으로 계통 분석을 실시한 결과, GYP-1 균주는 Rhodotorula 속에 속하며, GYP-3-3은 Brachybacterium 속에 속하는 것으로 확인되었다. 특히, Rhodotorula sp. GYP-1 균주는 호기성 효모종으로 biomass 생산량이 높아 최종 우수 균주로 선발하였다. Rhodotorula sp. GYP-1가 생성하는 𝛽-Glucosidase의 온도 및 pH에 따른 효소 활성 및 안정성을 검정한 결과, 30 ℃에서 6.7 unit/ml로 가장 높은 활성을 나타내었고, 20 ℃ ~ 40 ℃에서 효소 활성의 약 70 % 이상을 유지하는 것으로 확인하였다. pH에 따른 효소의 활성 및 안정의 경우, pH 5에서 6.8 unit/ml으로 가장 높은 활성을 나타내었고 pH 5~pH 8까지 93.3 % 이상의 효소 활성을 유지하는 것으로 확인하였다. Rhodotorula sp. GYP-1가 생성하는 𝛽-Glucosidase는 ginsenoside Rb1 minor 진세노사이드로 분해하는 것으로 확인되었다. 또한, 인삼 뿌리 병원균(Botrytis cinerea)에 대해 항진균능을 갖는 것으로 확인되었다.

• 한국어병학회지
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• 제36권2호
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• pp.251-261
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• 2023

Photobacterium sp. YW2207 was isolated from rainbow trout raised in a fish farm located in Yeongwol-gun, Gangwon Province, South Korea. Based on 16S rRNA sequence analysis and phylogenetic analysis, it was confirmed that Photobacterium sp. YW2207 showed 100% similarity with Photobacterium piscicola and Photobacterium phosphoreum, and 94.6% similarity with P. damselae subsp. damselae. Biochemical analysis revealed that Photobacterium sp. YW2207 is a Gram-negative, motile bacterium with a cell size of 1.5~3×3~5 ㎛. The bacteria were cultured on nutrient agar, brain heart infusion agar, Muller-Hinton agar, tryptic soy agar, and thiosulfate citrate bile sucrose agar with NaCl concentrations ranging from 0 to 2.5%. The API50CHE and API20E tests indicated lower utilization capabilities compared to the P. damselae strains provided in the API database. Furthermore, unlike most Photobacterium species, Photobacterium sp. YW2207 presented negative for catalase test. Results from the flow cytometric measurement indicated that Photobacterium sp. YW2207 exhibited a more diverse distribution of cell sizes and had larger cell sizes compared with P. damselae subsp. damselae. Minimum inhibitory concentration tests showed that Photobacterium sp. YW2207 had low susceptibility to β-Lactam and aminoglycoside antibiotics, while having high susceptibility to tetracycline, doxycycline, and quinolone antibiotics. Pathogenicity on rainbow trout revealed that an immersion of 1×10⁵ CFU/ml did not cause mortality or clinical symptoms.

• Journal of Ginseng Research
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• 제47권5호
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• pp.627-637
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• 2023

Background: Damage to the healthy intestinal epithelial layer and regulation of the intestinal immune system, closely interrelated, are considered pivotal parts of the curative treatment for inflammatory bowel disease (IBD). Plant-based diets and phytochemicals can support the immune microenvironment in the intestinal epithelial barrier for a balanced immune system by improving the intestinal microecological balance and may have therapeutic potential in colitis. However, there have been only a few reports on the therapeutic potential of plant-derived exosome-like nanoparticles (PENs) and the underlying mechanism in colitis. This study aimed to assess the therapeutic effect of PENs from Panax ginseng, ginseng-derived exosome-like nanoparticles (GENs), in a mouse model of IBD, with a focus on the intestinal immune microenvironment. Method: To evaluate the anti-inflammatory effect of GENs on acute colitis, we treated GENs in Caco2 and lipopolysaccharide (LPS) -induced RAW 264.7 macrophages and analyzed the gene expression of proinflammatory cytokines and anti-inflammatory cytokines such as TNF-α, IL-6, and IL-10 by real-time PCR (RT-PCR). Furthermore, we further examined bacterial DNA from feces and determined the alteration of gut microbiota composition in DSS-induced colitis mice after administration of GENs through 16S rRNA gene sequencing analysis. Result: GENs with low toxicity showed a long-lasting intestinal retention effect for 48 h, which could lead to effective suppression of pro-inflammatory cytokines such as TNF-α and IL-6 production through inhibition of NF-κB in DSS-induced colitis. As a result, it showed longer colon length and suppressed thickening of the colon wall in the mice treated with GENs. Due to the amelioration of the progression of DSS-induced colitis with GENs treatment, the prolonged survival rate was observed for 17 days compared to 9 days in the PBS-treated group. In the gut microbiota analysis, the ratio of Firmicutes/Bacteroidota was decreased, which means GENs have therapeutic effectiveness against IBD. Ingesting GENs would be expected to slow colitis progression, strengthen the gut microbiota, and maintain gut homeostasis by preventing bacterial dysbiosis. Conclusion: GENs have a therapeutic effect on colitis through modulation of the intestinal microbiota and immune microenvironment. GENs not only ameliorate the inflammation in the damaged intestine by downregulating pro-inflammatory cytokines but also help balance the microbiota on the intestinal barrier and thereby improve the digestive system.