• 한국자기공명학회논문지
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• 제22권4호
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• pp.149-157
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• 2018

Diabetes is known to be one of common causes for several types of peripheral nerve damage. Diabetic neuropathy (DN) is a significant complication lowering the quality of life that can be frequently found in diabetes patients. In this study, the metabolomic characteristic of DN and Diabetes was investigated with NMR spectroscopy. The sera samples were collected from DN patients, Diabetes patients, and healthy volunteers. Based on the pair-wise comparison, three metabolites were found to be noticeable: glucose, obviously, was upregulated both in DN patients (DNP) and Diabetes. Citrate is also increased in both diseases. However, the dietary nutrient and biosynthesized metabolite from glucose, ascorbate, was elevated only in DNP, compared to healthy control. The multivariate model of OPLS-DA clearly showed the group separation between healthy control-DNP and healthy control-Diabetes. The most significant metabolites that contributed the group separation included glucose, citrate, ascorbate, and lactate. Lactate did not show the statistical significance of change in t-test while it tends to down-regulated both in DNP and Diabetes. We also conducted the ROC curve analysis to make a multivariate model for discrimination of healthy control and diseases with the identified three metabolites. As a result, the discrimination model between healthy control and DNP (or Diabetes) was successful while the model between DNP and Diabetes was not satisfactory for discrimination. In addition, multiple combinations of lactate and citrate in the OPLS-DA model of healthy control and diabetes group (DNP + Diabetes patients) gave good ROC value of 0.952, which imply these two metabolites could be used for diagnosis of Diabetes without glucose information.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1144-1154
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• 2019

There have been several studies regarding lichen-associated bacteria obtained from diverse environments. Our screening process identified 49 bacterial species in two lichens from the Himalayas: 17 species of Actinobacteria, 19 species of Firmicutes, and 13 species of Proteobacteria. We discovered five types of strong antimicrobial agent-producing bacteria. Although some strains exhibited weak antimicrobial activity, NP088, NP131, NP132, NP134, and NP160 exhibited strong antimicrobial activity against all multidrug-resistant strains. Polyketide synthase (PKS) fingerprinting revealed results for 69 of 148 strains; these had similar genes, such as fatty acid-related PKS, adenylation domain genes, PfaA, and PksD. Although the association between antimicrobial activity and the PKS fingerprinting results is poorly resolved, NP160 had six types of PKS fingerprinting genes, as well as strong antimicrobial activity. Therefore, we sequenced the draft genome of strain NP160, and predicted its secondary metabolism using antiSMASH version 4.2. NP160 had 46 clusters and was predicted to produce similar secondary metabolites with similarities of 5-100%. Although NP160 had 100% similarity with the alkylresorcinol biosynthetic gene cluster, our results showed low similarity with existing members of this biosynthetic gene cluster, and most have not yet been revealed. In conclusion, we expect that lichen-associated bacteria from the Himalayas can produce new secondary metabolites, and we found several secondary metabolite-related biosynthetic gene clusters to support this hypothesis.

• 미생물학회지
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• 제55권2호
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• pp.157-159
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• 2019

낙동강 주변에서 채취한 토양으로부터 분리한 Pseudomonas fluorescens NBC275 (Pf275) 균주는 식물과 곤충에 병을 일으키는 곰팡이류에 우수한 항균력을 보였다. 본 연구에서는 Pf275 균주의 전체염기서열을 해독하고 분석하였는데, 총 염기서열은 6,610,362 bp였고, GC 함량은 60.9%였다. 염색체는 5,869개의 단백질을 암호화하였고, 16개의 rRNA와 65개의 tRNA로 구성되어 있었다. 유전체의 분석을 통해 항균력을 나타내는 2차 대사산물을 암호화하는 유전자를 확인할 수 있었는데, Pf275 균주는 pyoverdine, 2, 4-diacetylphloroglucinol 및 phenazine 등의 항균물질을 생산하였고, 이들 대사산물에 의해 항균력 및 생물방제효과를 나타내는 것으로 판단된다.

• 한국미생물·생명공학회지
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• 제46권4호
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• pp.416-424
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• 2018

강원도에서 생산된 과일을 사용하여 식초를 제조하여, 초산 발효에 의하여 변화되는 주요 대사체 및 향기 성분을 조사하였다. 머루 복분자 식초에서는 초산 발효에 의하여 ${\text\tiny{L}}$-alanine의 함량 변화가 가장 크게 변화하였으며, 초산은 향기 성분 전체 함량 중 43%로 가장 높은 함량을 나타냈고, 초산 발효 후 ethyl acetate, isoamyl acetate 등의 에스테르 화합물의 함량이 증가하였다. 오미자 막걸리 식초의 경우 초산발효에 의하여 linalool, hexanoic acid 등이 생성되었고 테르펜류 화합물이 41.5%로 대부분을 차지하였다. 머루 복분자 식초의 경우와 유사하게 ${\text\tiny{L}}$-alanine의 함량 변화가 가장 큰 것으로 나타났다. 머루 복분자 식초에서는 ethyl acetate, 오미자 막걸리 식초에서는 감귤향을 내는 terpinel-4-ol, 꽃향기를 내는 ${\alpha}$-terpineol 등 테르펜류의 성분들이 각 식초의 향기 특성에 큰 영향을 나타내는 것으로 확인되었다.

• 분석과학
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• 제34권3호
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• pp.115-121
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• 2021

에탄올 대사체인 에틸글루쿠로나이드(ethyl glucuronide, EtG)와 에틸설페이트(ethyl sulfate, EtS)는 에탄올 섭취의 직접적인 지표물질로서 여러 생체 시료에서 법과학적으로 다양하게 연구되고 있다. 부검 변사자에게서 검출된 에탄올이 사망 전 음주에 의한 것인지 사후 부패에 의해 생성된 에탄올인지 판단하기 위해서는 에탄올 대사체를 분석할 필요가 있다. 일반적으로 혈액에서 에탄올과 함께 EtG, EtS를 검출하고 있지만 부패 상황에 따라 혈액을 확보하는 것이 어려우므로 조직 생체시료를 이용해 에탄올 대사체를 검출함으로써 이를 대체해야 한다. 본 연구에서는 조직 중에서 비장을 선택하여 LC-MS/MS를 이용해 EtG, EtS를 검출하였으며 시험법이 적합한 방법인지를 증명하기 위해 밸리데이션을 수행하여 법과학적 감정에 사용할 수 있는 분석법을 완성하고 실제 부검 사례에 적용하였다.

• Journal of Ginseng Research
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• 제45권1호
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• pp.98-107
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• 2021

Background: Ginseng extracts and ginseng-fermented products are widely used as functional cosmetic ingredients for their whitening and antiwrinkle effects. Recently, increasing attention has been given to bioactive metabolites isolated from endophytic fungi. However, little is known about the bioactive metabolites of the fungi associated with Panax ginseng Meyer. Methods: An endophytic fungus, Penicillium sp. SNF123 was isolated from the root of P. ginseng, from which acremonidin E was purified. Acremonidin E was tested on melanin synthesis in the murine melanoma cell line B16F10, in the human melanoma cell line MNT-1, and in a pigmented 3D-human skin model, Melanoderm. Results: Acremonidin E reduced melanogenesis in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 cells with minimal cytotoxicity. qRT-PCR analysis demonstrated that acremonidin E downregulated melanogenic genes, including tyrosinase and tyrosinase-related protein 1 (TRP-1), while their enzymatic activities were unaffected. The antimelanogenic effects of acremonidin E were further confirmed in MNT-1 and a pigmented 3D human epidermal skin model, Melanoderm. Immunohistological examination of the Melanoderm further confirmed the regression of both melanin synthesis and melanocyte activation in the treated tissue. Conclusion: This study demonstrates that acremonidin E, a bioactive metabolite derived from a fungal endophyte of P. ginseng, can inhibit melanin synthesis by downregulating tyrosinase, illuminating the potential utility of microorganisms associated with P. ginseng for cosmetic ingredients.

• Journal of Applied Biological Chemistry
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• 제65권3호
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• pp.195-202
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• 2022

콩잎의 기능성식품소재연구가 활발히 진행되고 있지만 기능성 식품소재로서 콩잎을 관리할 수 있는 밸리데이션 방법은 제시되지 않고있다. 본 연구에서는 콩잎에서 7개의 kaempferol 유도체를 UHPLC-ESI-Q-TOF-MS로 동정하고 이들 중 camelliaside A를 지표물질로 선정하였다. 콩잎열수추출물에서 camelliaside A를 정량할 수 있는 HPLC-UVD 밸리데이션 방법을 식품의약품안전처의 건강기능성식품에 대한 가이드라인에 따라 구축하였다. 개발된 HPLC-UVD 밸리데이션 방법은 특이성, 정확도, 일내정밀도, 일간정밀도, 검출한계, 정량한계 및 직선성에서 가이드라인을 만족하는 결과값으로 검증되었다. 또한 camelliaside A 분석법을 콩잎열수추출물에 적용하여 적합한 수준의 평가값을 얻었다.

• Journal of Ginseng Research
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• 제47권2호
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• pp.311-318
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• 2023

Background: The beneficial effects of compound K (CK) on different chronic diseases have been shown to be at least related to antioxidant action. Nevertheless, since its antioxidant activity in human retinal pigment epithelial (RPE) cells is still unknown, here we investigated whether CK alleviates oxidative stress-stimulated damage in RPE ARPE-19 cells. Methods: The cytoprotective consequence of CK in hydrogen peroxide (H₂O₂)-treated cells was evaluated by cell viability, DNA damage, and apoptosis assays. Fluorescence analysis and immunoblotting were performed to investigate the inhibitory action of CK on reactive oxygen species (ROS) production and mitochondrial dysfunction. Results: H₂O₂-promoted cytotoxicity, oxidative stress, DNA damage, mitochondrial impairment, and apoptosis were significantly attenuated by CK in ARPE-19 cells. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation level and its shuttling to the nucleus were increased, which was correlated with upregulated activation of heme oxygenase-1 (HO-1). However, zinc protoporphyrin, a blocker of HO-1, significantly abrogated the preventive action of CK in H₂O₂-treated ARPE-19 cells. Conclusion: This study indicates that activation of Nrf2/HO-1 signaling by CK plays an important role in rescuing ARPE-19 cells from oxidative cellular damage.

• Journal of Microbiology and Biotechnology
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• 제33권6호
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• pp.806-822
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• 2023

In the current study we assessed a new crystallized compound, 5-(1-hydroxybutyl)-4-methoxy-3-methyl-2H-pyran-2-one (C-HMMP), from the endophytic fungus Colletotrichum acutatum residing in the medicinal plant Angelica sinensis for its in vitro antimicrobial, antibiofilm, antioxidant, antimalarial, and anti-proliferative properties. The promising compound was identified as C-HMMP through antimicrobial-guided fraction. The structure of C-HMMP was unambiguously confirmed by 2D NMR and HIRS spectroscopic analysis. Antimicrobial property testing of C-HMMP showed it to be effective against a variety of pathogenic bacteria and fungi with MICs ranging from 3.9 to 31.25 ㎍/ml. The compound displayed excellent antibiofilm activity against C. albicans, S. aureus, and K. pneumonia. Furthermore, the antimalarial and radical scavenging activities of C-HMMP were clearly dosedependent, with IC50 values of 0.15 and 131.2 ㎍/ml. The anti-proliferative activity of C-HMMP against the HepG-2, HeLa, and MCF-7 cell lines in vitro was investigated by MTT assay, revealing notable anti-proliferative activity with IC50 values of 114.1, 90, and 133.6 ㎍/ml, respectively. Moreover, CHMMP successfully targets topoisomerase I and demonstrated beneficial anti-mutagenicity in the Ames test against the reactive carcinogenic mutagen, 2-aminofluorene (2-AF). Finally, the compound inhibited the activity of α-glucosidase and α-amylase with IC50 values of 144.7 and 118.6 ㎍/ml, respectively. To the best of our knowledge, the identified compound C-HMMP was obtained for the first time from C. acutatum of A. sinensis, and this study demonstrated that C-HMMP has relevant biological significance and could provide better therapeutic targets against disease.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제26권2호
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• pp.99-115
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• 2023

Purpose: Exclusive breastfeeding promotes gut microbial compositions associated with lower rates of metabolic and autoimmune diseases. Its cessation is implicated in increased microbiome-metabolome discordance, suggesting a vulnerability to dietary changes. Formula supplementation is common within our low-income, ethnic-minority community. We studied exclusively breastfed (EBF) neonates' early microbiome-metabolome coupling in efforts to build foundational knowledge needed to target this inequality. Methods: Maternal surveys and stool samples from seven EBF neonates at first transitional stool (0-24 hours), discharge (30-48 hours), and at first appointment (days 3-5) were collected. Survey included demographics, feeding method, medications, medical history and tobacco and alcohol use. Stool samples were processed for 16S rRNA gene sequencing and lipid analysis by gas chromatography-mass spectrometry. Alpha and beta diversity analyses and Procrustes randomization for associations were carried out. Results: Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria were the most abundant taxa. Variation in microbiome composition was greater between individuals than within (p=0.001). Palmitic, oleic, stearic, and linoleic acids were the most abundant lipids. Variation in lipid composition was greater between individuals than within (p=0.040). Multivariate composition of the metabolome, but not microbiome, correlated with time (p=0.030). Total lipids, saturated lipids, and unsaturated lipids concentrations increased over time (p=0.012, p=0.008, p=0.023). Alpha diversity did not correlate with time (p=0.403). Microbiome composition was not associated with each samples' metabolome (p=0.450). Conclusion: Neonate gut microbiomes were unique to each neonate; respective metabolome profiles demonstrated generalizable temporal developments. The overall variability suggests potential interplay between influences including maternal breastmilk composition, amount consumed and living environment.