• 생명과학회지
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• 제17권12호
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• pp.1649-1653
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• 2007

무더운 날씨가 지속됨으로서 고랭지배추의 생장 및 결구가 지연되고 있는 강원도 정선군 질운산(새빗재)의 600 m와 900 m의 배추를 사용하여 무기성분 및 단백질 발현패턴을 분석하였다. 식물체 무기성분에서는 생장에 관련된 질소 및 인산의 부족현상과 결구에 관련된 칼슘이 부족하였다. 단백체 분석은 2차원 전기영동에 의해 전체 126개의 단백질이 분리되었고 그중 48개의 단백질이 고도에 따라 변화하는 양상을 보여주었다. 이 중에서 30개의 단백질 서열이 결정되었는데, 해발 900 m에서 단백질 발현이 증가한 14개 중에서 oxygen- evolving proteins, rubisco activase and ATPase 등이, 해발 600 m에서는 glutathione S-transferase (1, 28 kD cold induced- and 24kD auxin-binding proteins) and salt-stress induced protein 등 16개의 단백질 발현이 증가하였다. 이러한 단백질은 식물체 손상에 대한 보호기작을 가진 스트레스관련 단백질로 가뭄, 온도상승, 밤낮의 온도차 등의 반복으로 복합적이며 동시 다발적으로 나타나는 고온장해 현상으로 사료된다.

• 한국물환경학회지
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• 제23권5호
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• pp.705-711
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• 2007

Green quantum-dot nanocrystal (QD525) with anti-microcystin monoclonal antibody was applied for detection of microcystin, a monocyclic peptide hepatotoxin, extracted from the culture of Microcystis aeruginosa. The presence of microcystin in the cell lysate was verified by HPLC analysis with UV absorbance at 238 nm. Microcystis cell extract exhibited fluorescence emission spectra, which peak was around 460 nm because of their complex organic substances. When a spherical QD525 antibody conjugates (10~20 nm in diameter) were bound to the microcystins in the Microcystis cell lysate, the fluorescence intensity of the primary peak at 525 nm diminished while the secondary emission peak at 460 nm slightly increased intensities. It is due to energy transfer from the primary (major) to the secondary (minor) peak, resulting from physical deformation of QD525 and different environmental factors. On the other hand, other cell extracts did not show any fluorescence emission change. This study is very available for detecting and monitoring the microcystin because it is one step assay without washing step and portable spectrophotometer makes on-site measurement possible. For health risk assessment of the microcystin, the reliable and rapid system to detect and quantify microcystin is seriously required.

• KSBB Journal
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• 제25권2호
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• pp.123-129
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• 2010

An endospore-forming, rod-shaped bacterium was isolated from forest soil samples collected at the Taebaek mountain of Gangwon province, Korea, and taxonomically characterized by physiological, biochemical and phylogenetic methods. Its 16S rRNA sequences showed the maximum similarity of 97% with B. amyloliquefaciens. In addition, the isolate BCNU 2002 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase. The in vitro antifungal activity of Bacillus sp. BCNU 2002 was also examined against human pathogenic fungi such as Aspergillus niger, Candida albicans, Epidermophyton floccosum, Saccharomyces cerevisiae, Trichophyton mentagrophytes and Trichophyton rubrum. A maximum production level of antifungal substances of Bacillus sp. BCNU 2002 was achieved under aerobic incubation at $28^{\circ}C$ for 7 days in LB broth. BCNU 2002 showed strong antifungal activities against T. mentagrophytes and T. rubrum with the range of percentage inhibition from 56.25 to 63.23%. It was also confirmed that ethylacetate extract of cultured broth showed a strong antifungal activity against A. niger, C. albicans, S. cerevisiae and T. rubrum by agar diffusion method. The peptide fraction also exhibited broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration values for active extracts ranged between 125 ${\mu}g$/mL and 1000 ${\mu}g$/mL.

• 한국수산과학회지
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• 제50권1호
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• pp.85-91
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• 2017

Alzheimer's disease (AD) is a progressive neurodegenerative disorder of insidious onset that causes gradual loss of memory and cognitive function, and it is the most common form of dementia in the elderly. AD is characterized by neuritic plaques and neurofibrillary tangles in the brain, together with loss of neuronal cells. The major neuropathological hallmark of AD is the accumulation of extracellular neurotoxic ${\beta}-amyloid$ ($A{\beta}$) peptides, such as $A{\beta}1-42$, in the brain. In the present study, we investigated the effect of sargachromenol (SCM), sargaquinoic acid (SQA) and sargahydroquinoic acid (SHQA) isolated from Sargassum serratifoilum ethanol extract (SSE) on $A{\beta}$ production in vitro using APP751-transfected Chinese hamster ovary cells (CHO-751). CHO-751 cells were treated with various concentrations of SSE, SCM, SQA and SHQA, and the level of extracellular $A{\beta}1-42$ was evaluated by enzyme-linked immunosorbent assay. SSE and SHQA reduced the production of $A{\beta}1-42$ in CHO-751 cells. Therefore, SHQA isolated from S. serratifolium has potential as an inhibitor of neurotoxic $A{\beta}$ peptide production.

• 미생물학회지
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• 제45권1호
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• pp.54-57
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• 2009

대두발효식품인 청국장에는 다양한 항산화물질, peptide 등의 생리활성물질이 존재한다. 청국장 ethanol 추출물은 285 nm에서 0.55의 흡광도 값을 보여 주었다. 이 영역에는 phenol을 포함한 아미노산 및 peptide류가 포함되어 있다고 알려져 있다. 1,1-diphenyl-2-picrylhydrazyl (DPPH)방법으로 청국장의 항산화도를 측정했을 때, ethanol 추출물의 농도가 증가할수록 항산화도는 증가하였다. 무염 생청국장 30 g을 복용하고 2시간 마다 혈압을 측정하였다. 수축기혈압은 복용 후 6시간 지난 후, 평균혈압이 14 mmHg 떨어졌고, 이완기혈압은 8 mmHg 떨어지는 등 혈압강하 효과가 뚜렷하였다. 청국장에 존재하는 daidzein, 항산화물질, Lys-Pro과 같은 angiotensin I-converting enzyme (ACE) 억제제 등이 복합적으로 작용하면서 혈압강하에 기여할 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1384-1391
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• 2006

The signal transduction system is one of the most important devices involved in maintaining life, and many protein kinases are included in the cellular signal transduction system. Finding a protein kinase inhibitor is very valuable, as it can be used to study cell biology and applied to pharmaceuticals. For the efficient and rapid screening of protein kinase inhibitors, two assay systems were combined; the nonradioactive protein kinase assay system that uses an FITC-labeled IRS-2 peptide and the cell-based paper disc assay system that uses Streptomyces griseus as the indicator strain. Among 330 kinds of herb extracts tested, the extract of Dalbergia odorifera exhibited the strongest inhibitory activity in the two assay systems and was selected for further isolation. Based on solvent extraction and many steps of chromatography, seven compounds were finally separated to homogeneity and their structures determined by $^{1}H$ and $^{13}C$ NMR spectroscopies. Four were to be flavonoids and identified as butin ($C_{15}H_{12}O_5$, Mw=272.07), 3'-hydroxymelanetin ($C_{16}H_{12}O_6$, Mw=300.06), liquiritigenin ($C_{15}H_{12}O_4$, Mw=256.07), and 2'-hydroxyformononetin ($C_{16}H_{12}O_{5}$, Mw=284.07). 3'-Hydroxymelanetin inhibited the phosphorylation of the GSK3 protein by Akt to 37% at a concentration of $10{\mu}g/ml$ and showed the strongest cytotoxicity ($ED_{50}<50{\mu}g/ml$) against the human cancer cell line HCT116. Under the same conditions, liquiritigenin also inhibited the phosphorylation of GSK3 by Akt to 26%, and its cytotoxicity against the HCT116 cell line was lower than $100{\mu}g/ml$.

• International Journal of Industrial Entomology and Biomaterials
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• 제37권2호
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• pp.95-99
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• 2018

In insect defense system, antimicrobial peptides (AMPs) are one of important biological molecules to survive in a variety of environments. Insect can synthesize AMPs to protect against invading pathogens in humoral immune response. Taking more advantage of biological antimicrobial molecules, we report antibacterial activity of two cecropin type peptides, cecropin and moricin, isolated from the silkworm against four salmonella species. In this work, we purified antimicrobial candidate peptides (AMCP) from the extracts of immune challenged silkworm larval hemolymph by two-step chromatographic purification procedure, cation exchange and gel permeation chromatography. The molecular weights of purified peptides were estimated to be about 4 ~ 5 kDa by Tricin SDS-PAGE analysis, and identified as silkworm cecropin and moricin by NCBI BLAST homology search with their N-terminal amino acid sequences. As antibacterial activity assay, the purified peptides showed stronger antibacterial activity against Salmonella pathogens with an MIC value of $1{\sim}4{\mu}g/mL$. Therefore two cecropin type peptides purified from the silkworm will be valuable potential materials for development of new natural antibiotics.

• 농업과학연구
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• 제48권3호
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• pp.557-565
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• 2021

Interest in research on various medicinal plants has increased globally over the last few decades, possibly due to their possible antibacterial and antioxidant activities. The present study was conducted to verify the antioxidant effects, antibacterial activity, and collagen synthesis and cell viability outcomes of adipocytes upon exposure to Abeliophyllum distichum Nakai (AdN). Antibacterial activity was measured through the Disc diffusion method to compare the growth ability of pathogenic microorganisms (E.coli, Salmonella). The absorbance was measured at 560 nm to calculate the active oxygen scavenging ability. Fibroblasts were dispensed in a 96-well plate at a density of 1 × 10⁵ cells·well^-1. The amount of procollagen was measured in each case using a procollagen type 1 C-peptide EIA KIT. The cytotoxicity of the Abeliophyllum distichum Nakai extract against animal adipocytes (Hanwoo backfat cells) was determined using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, a method that measures the conversion of MTS to Formazan by means of mitochondrial dehydrogenases. The concentrations of the samples were made to be 0.0125, 0025, 0.05, 0.1, and 0.5% and all were -completely absorbed into the disc in an incubator at 37℃ for 24 to 36 hours. For the 0.125 mg·disc^-1, effects of Abeliophyllum distichum Nakai on the antioxidant effect, antibacterial activity, and cell viability of adipocytes were found. However, Abeliophyllum distichum Nakai had no effect on collagen synthesis, thus suggesting that AdN extracts may be useful for the prevention and/or treatment of obesity.

• Journal of Dairy Science and Biotechnology
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• 제41권3호
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• pp.113-125
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• 2023

In this study, we explored the synergistic effects of whey protein concentrate (WPC) and soybean protein components after fermentation with lactic acid bacteria isolated from kimchi, and identified several peptides with desirable physiological functions, proteolysis, and immune effects. Antioxidant activity was determined using 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid, 1,1-diphenyl-2-picrylhydrazyl, ferric-reducing antioxidant power, and hydroxyl radical scavenging assays, followed by cross-validation of the four antioxidant activities. These assays revealed that samples with a 8:2 and 9:1 whey to soy ratio possessed higher antioxidant activity than the control samples. Antibacterial potency testing revealed high antibacterial activity in the 9:1 and 8:2 samples. Cytotoxicity testing of samples using 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide revealed that only the 10:0, 1:9, and 0:10 samples had <80% viable cells, indicating no significant cytotoxicity. Nitric oxide (NO) assays revealed that NO expression was reduced in 8:2, 5:5, and 0:10 protein ratio fermentations, indicating low inflammatory reaction stimulatory potential. Cytokine expression was confirmed using an enzyme-linked immunosorbent assay kit. The 8:2 sample had the lowest inflammatory cytokine (interleukin [IL]-1α, IL-6, and tumor necrosis factor-α) levels compared with the lipopolysaccharide-treated group. Amino acid profiling of the 8:2 sample identified 17 amino acids. These results suggest that inoculating and fermenting Lactobacillus plantarum DK203 and Lactobacillus paracasei DK209 with an 8:2 mixture of WPC and soybean protein releases bioactive peptides with excellent anti-inflammatory and antioxidant properties, making them suitable for functional food development.

• KSBB Journal
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• 제22권5호
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• pp.356-361
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• 2007

시아노박테리아 같은 수중 미생물에 대한 2차대사물질에 대한 연구는 육상식물이나 미생물에 관련된 연구방법을 응용하고 있으며 아직까지 체계화 되어있지 않아 시아노박테리아를 보다 효율적으로 조사하기 위한 새로운 연구기술의 모색이 절실히 필요하다. 이 연구에서는 의학적인 관점에서 시아노박테리아를 조사하기 위한 유용한 접근 방법을 모색하였고 체계화시켰다. 균주마다 특성화된 최적 배양을 하였고 PCR 증폭을 이용한 분자생물학적인 방법과 HPLC를 통한 정성분석으로 의학적으로 의미있는 NPs를 생산하는 유망 균주를 선별하였다. 선별된 균주에서 나온 추출액은 SPE와 preparative HPLC를 거쳐 분리 정제되어지고, 정제된 물질들은 질량분석기에 의해 분자량과 구조가 결정되어 졌으며, 생활성 테스트에 의해서 그 생물학적인 활성을 정함으로써 의학적인 가능성이나 활용성에 대해서 고찰하였다. 이번 시험에서 98개의 실험균주 중 46개의 균주가 NRPS 또는 PKS 관련 gene을 함유하고 있었으나 HPLC를 이용한 정성분석에서 단지 5개의 균주가 상당히 의미있는 관련 단백질을 생산하고 있음이 알려졌다. 즉, 41 균주에서 관련 gene은 존재하였지만 그 발현이 미약하였고 또는 프로모터의 활성이 이루어지지 않았다. 이와같은 휴먼 gene의 활성화는 자외선 조사 또는 건조를 이용한 자극과 배양액의 성분 조절로 이루어질 수 있고 이를 통해 흥미있는 결과를 이끌어낼 수 있다(13, 14). 이번 연구에서는 HPLC를 통한 정성분석은 Biomass에 대해서 이루어졌다. 그러므로 배양액으로 유출되는 extracellular substances에 대한 분석은 행해지지 않았다. 시아노박테리아가 NRPS/PKS 관련 유전자를 함유하고 있을 때, 특히 biomass에서 NPs 이 검출되지 않을 경우 배양액 안에서의 존재 가능성 대해서 조사할 필요성이 요구되어진다. 시아노박테리아는 대표적인 photoautotroph 미생물로써 $CO_2$를 탄소성분과 에너지원으로 사용한다. 그러므로 E. coli 같은 미생물보다 배양시 경제적이고 또한 다른 종류의 미생물이 탄소영양분의 부족과 cyanobacterial NPs의 생물학적 환성에 의해서 공생하기 어려워 옥외배양을 통한 대량생산이 가능하다. 그러나, 생산되는 cyanobacterial NPs의 높은 구조적 안정성까지 포함하여 시아노박테리아는 미래의학산업의 중요한 천연소스임에 틀림이 없지만 개체 분화의 시간이 4$\sim$10시간 정도로 다른 미생물에 비해 길고 배양시 광원의 필요성 등 한계 요소를 지니고 있다(15). 그러므로 최근에는 개체 분화가 빠른 시아노박테리아나 또는 다른 미생물에 대해서 cyanobacterial NRPS/PKS gene의 heterologous expression 이 연구되어지고 있다(16).