• 한국식품저장유통학회지
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• 제18권6호
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• pp.973-979
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• 2011

Carbonic maceration 처리는 포도주 제조 시 사과산을 감소시키는 방법으로 사과산 감소의 원인 중 미생물의 영향을 알아보고자 사과산을 감소시키고 젖산을 생성시키는 미생물을 분리, 동정한 결과Lactobacillus brevis, Lactobacillus plantarum 및 Streptococcus thermophilus의 젖산균이 존재하는 것으로 나타났다. 분리된 균들은 대부분 당에서 젖산을 생성하는 균으로 알려져 있으며 사과산을 함유한 배지에서 균의 배양 중 사과산을 이용하지 않고 젖산을 생성하는 것으로 보아 주로 당을 이용하여 젖산을 생성하는 것으로 보인다. 사과산을 이용해 젖산을 생성시키는 대표적인 malo-lactic bacteria인 Oenococcus oeni 균은 본 실험에서는 동정되지 않았다. 따라서 carbonic maceration 처리 시 사과산의 감소는 포도에 자연적으로 생육한다는 malo-lactic bacteria나 감산 관련 미생물의 영향은 크게 받지 않는 것으로 판단되며, 젖산 함량의 증가는 당을 이용하는 다양한 젖산균에 의해 생성되는 것으로 판단된다.

• Journal of Animal Science and Technology
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• 제63권4호
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• pp.934-953
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• 2021

Ciglitazone is a member of the thiazolidinedione family, and specifically binds to peroxisome proliferator-activated receptor-γ (PPARγ), thereby promoting adipocyte differentiation. We hypothesized that ciglitazone as a PPARγ ligand in the absence of an adipocyte differentiation cocktail would increase adiponectin and adipogenic gene expression in bovine satellite cells (BSC). Muscle-derived BSCs were isolated from six, 18-month-old Yanbian Yellow Cattle. The BSC were cultured for 96 h in differentiation medium containing 5 µM ciglitazone (CL), 10 µM ciglitazone (CM), or 20 µM ciglitazone (CH). Control (CON) BSC were cultured only in a differentiation medium (containing 2% horse serum). The presence of myogenin, desmin, and paired box 7 (Pax7) proteins was confirmed in the BSC by immunofluorescence staining. The CL, CM, and CH treatments produced higher concentrations of triacylglycerol and lipid droplet accumulation in myotubes than those of the CON treatment. Ciglitazone treatments significantly increased the relative expression of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), C/EBPβ, fatty acid synthase, stearoyl-CoA desaturase, and perilipin 2. Ciglitazone treatments increased gene expression of Pax3 and Pax7 and decreased expression of myogenic differentiation-1, myogenin, myogenic regulatory factor-5, and myogenin-4 (p < 0.01). Adiponectin concentration caused by ciglitazone treatments was significantly greater than CON (p < 0.01). RNA sequencing showed that 281 differentially expressed genes (DEGs) were found in the treatments of ciglitazone. DEGs gene ontology (GO) analysis showed that the top 10 GO enrichment significantly changed the biological processes such as protein trimerization, negative regulation of cell proliferation, adipocytes differentiation, and cellular response to external stimulus. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that DEGs were involved in the p53 signaling pathway, PPAR signaling pathway, biosynthesis of amino acids, tumor necrosis factor signaling pathway, non-alcoholic fatty liver disease, PI3K-Akt signaling pathway, and Wnt signaling pathway. These results indicate that ciglitazone acts as PPARγ agonist, effectively increases the adiponectin concentration and adipogenic gene expression, and stimulates the conversion of BSC to adipocyte-like cells in the absence of adipocyte differentiation cocktail.

• 한국자원식물학회지
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• 제34권4호
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• pp.278-286
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• 2021

유전자 가위 기술 등 생명공학 기술을 콩에 적용하여 새로운 품종을 개발하기 위해서는 효율적인 조직배양 기술이 필수적이다. 식물의 유전형은 조직배양 효율에 의존하는 형질전환 기술의 성공 여부를 결정짓는 중요한 요소로 알려져 있다. 본 연구에서는 우리나라 콩 핵심 집단 내 21개 자원들을 선발하여, 외래 품종 2종과 함께 조직배양 효율을 조사하였다. 그 결과, 근연 관계가 높은 Kwangan, Anpyeong, Seonam은 발아율과 재분화 효율이 높았으며, Daepung, Daewon 품종은 발아율과 재분화율 모두 낮게 관찰되었다. 또한 3종의 외래 품종에서는 표준 유전체 해독에 사용된 Williams82와 Jack, Maverick 모두 높은 조직배양 효율을 보였다. 조직배양 효율이 높은 자원들을 대상으로 Agrobacterium법에 의한 형질전환을 수행하여 PCR 및 bar-strip 분석한 결과 Kwangan, Pungwon, Seonam, 그리고 Maverick 품종에서 제초제 저항성 유전자의 삽입을 확인할 수 있었다. 이들 결과를 바탕으로 농업적 가치가 높은 다양한 콩 품종들의 형질전환을 통한 새로운 품종 개발이 가능할 것이다.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1083-1095
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• 2019

Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on $NH_3-N$ at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, $NH_3-N$ and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with $10^6CFU/ml$ C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

• BMB Reports
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• 제52권7호
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• pp.463-468
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• 2019

Autosomal dominant polycystic kidney disease (ADPKD), one of the most common human monogenic diseases (frequency of 1/1000-1/400), is characterized by numerous fluid-filled renal cysts (RCs). Inactivation of the PKD1 or PKD2 gene by germline and somatic mutations is necessary for cyst formation in ADPKD. To mechanistically understand cyst formation and growth, we isolated RCs from Korean patients with ADPKD and immortalized them with human telomerase reverse transcriptase (hTERT). Three hTERT-immortalized RC cell lines were characterized as proximal epithelial cells with germline and somatic PKD1 mutations. Thus, we first established hTERT-immortalized proximal cyst cells with somatic PKD1 mutations. Through transcriptome sequencing and Gene Ontology (GO) analysis, we found that upregulated genes were related to cell division and that downregulated genes were related to cell differentiation. We wondered whether the upregulated gene for the chemokine CXCL12 is related to the mTOR signaling pathway in cyst growth in ADPKD. CXCL12 mRNA expression and secretion were increased in RC cell lines. We then examined CXCL12 levels in RC fluids from patients with ADPKD and found increased CXCL12 levels. The CXCL12 receptor CXC chemokine receptor 4 (CXCR4) was upregulated, and the mTOR signaling pathway, which is downstream of the CXCL12/CXCR4 axis, was activated in ADPKD kidney tissue. To confirm activation of the mTOR signaling pathway by CXCL12 via CXCR4, we treated the RC cell lines with recombinant CXCL12 and the CXCR4 antagonist AMD3100; CXCL12 induced the mTOR signaling pathway, but the CXCR4 antagonist AMD3100 blocked the mTOR signaling pathway. Taken together, these results suggest that enhanced CXCL12 in RC fluids activates the mTOR signaling pathway via CXCR4 in ADPKD cyst growth.

• Animal Bioscience
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• 제34권2호
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• pp.243-255
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• 2021

Objective: Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that frequently contaminate maize and grain cereals, imposing risks to the health of both humans and animals and leading to economic losses. The gut microbiome has been shown to help combat the effects of such toxins, with certain microorganisms reported to contribute significantly to the detoxification process. Methods: We examined the cecum contents of three different dietary groups of pigs (control, as well as diets contaminated with 8 mg DON/kg feed or 0.8 mg ZEN/kg feed). Bacterial 16S rRNA gene amplicons were acquired from the cecum contents and evaluated by next-generation sequencing. Results: A total of 2,539,288 sequences were generated with ~500 nucleotide read lengths. Firmicutes, Bacteroidetes, and Proteobacteria were the dominant phyla, occupying more than 96% of all three groups. Lactobacillus, Bacteroides, Megasphaera, and Campylobacter showed potential as biomarkers for each group. Particularly, Lactobacillus and Bacteroides were more abundant in the DON and ZEN groups than in the control. Additionally, 52,414 operational taxonomic units were detected in the three groups; those of Bacteroides, Lactobacillus, Campylobacter, and Prevotella were most dominant and significantly varied between groups. Hence, contamination of feed by DON and ZEN affected the cecum microbiota, while Lactobacillus and Bacteroides were highly abundant and positively influenced the host physiology. Conclusion: Lactobacillus and Bacteroides play key roles in the process of detoxification and improving the immune response. We, therefore, believe that these results may be useful for determining whether disturbances in the intestinal microflora, such as the toxic effects of DON and ZEN, can be treated by modulating the intestinal bacterial flora.

• 생명과학회지
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• 제31권8호
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• pp.761-770
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• 2021

본 연구에서는 반응표면분석법을 이용하여 전통발효식품인 막걸리로부터 분리한 Bacillus amyloliquefaciens SRCM115785 균주에 대하여 protease 생산량을 증가시키기 위한 배지의 최적 농도를 확립하고자 하였다. 선정한 11개의 배지 성분 중 각 성분이 protease 생산에 미치는 영향에 대한 분석을 위해 Plackett-Burman design (PBD)를 설계하여 통계분석한 결과 glucose, yeast extract, beef extract를 protease 생산 향상을 위한 요인으로 최종 선별하였다. 선별된 3개의 성분에 대해 protease 생산을 위한 각 성분별 최적 농도를 결정하기 위해 central composite design (CCD)분석을 설계하여 protease 최대 생산을 위한 각 배지조성별 농도는 glucose 6.75 g/l, yeast extract 12.42 g/l, beef extract 17.48 g/l로 예측되었다. ANOVA 분석을 통해 실험모델의 적합성을 증명하였고, 설계한 최적배지에서 반복실험을 진행하여 protease 생산량을 측정한 결과 예측값과 매우 유사한 값을 나타냄을 확인하였다. 최종적으로 일반 배지에 비해 137% 환이 증가하였으며, 추가로 정량 분석 결과 기존 25.72 U/ml 대비 59.28 U/ml로 230.47% 증가함을 확인하였다. 본 연구를 통해 protease 생산량 증가를 위한 배지 성분의 최적화를 확립하였고, 이를 바탕으로 산업용 효소로서 protease의 효율적인 활용방안에 대한 기초자료로서 활용될 수 있을 것으로 기대된다.

• 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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• 제32권3호
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• pp.294-301
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• 2022

In our greenhouse experiment, soil heat treatment groups (50, 80, and 121℃) significantly promoted growth and disease suppression of Panax notoginseng in consecutively cultivated soil (CCS) samples (p < 0.01), and 80℃ worked better than 50℃ and 121℃ (p < 0.01). Furthermore, we found that heat treatment at 80℃ changes the microbial diversity in CCS, and the inhibition ratios of culturable microorganisms, such as fungi and actinomycetes, were nearly 100%. However, the heat-tolerant bacterial community was preserved. The 16S rRNA gene and internal transcribed spacer (ITS) sequencing analyses indicated that the soil heat treatment had a greater effect on the Chao1 index and Shannon's diversity index of bacteria than fungi, and the relative abundances of Firmicutes and Proteobacteria were significantly higher than without heating (80 and 121℃, p < 0.05). Soil probiotic bacteria, such as Bacillus (67%), Sporosarcina (9%), Paenibacillus (6%), Paenisporosarcina (6%), and Cohnella (4%), remained in the soil after the 80℃ and 121℃ heat treatments. Although steam increased the relative abundances of most of the heat-tolerant microbes before sowing, richness and diversity gradually recovered to the level of CCS, regardless of fungi or bacteria, after replanting. Thus, we added heat-tolerant microbes (such as Bacillus) after steaming, which reduced the relative abundance of pathogens, recruited antagonistic bacteria, and provided a long-term protective effect compared to the steaming and Bacillus alone (p < 0.05). Taken together, the current study provides novel insight into sustainable agriculture in a consecutively cultivated system.

• 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.