• 한국산학기술학회논문지
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• 제9권3호
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• pp.800-806
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• 2008

자연발효된 청국장으로부터 혈전용해활성을 가지는 세균 Bacillus licheniformis HK-12를 농화분리하여, 배양기간 동안 생산된 혈전용해활성을 가지는 단백질에 대해 프로테옴 분석을 실시하였다. B. licheniformis HK-12를 액체 영양배지에 접종하여 얻어진 배양상등액을 피브린 평판법(fibrin plate method)을 사용하여 효소활성을 측정하였다. 그 결과, HK-12의 혈전용해 활성은 대조구인 plasmin보다 약 2.3배정도 높은 활성을 나타내었다. 효소는 배양상등액을 ammonium sulfate 침전, DEAE-cellulose chromatography, Sephadex chromatography 등을 수행하여 분리정제하였으며, 정제된 혈전용해효소의 분자량은 SDS-PAGE를 통해 약 23 kDa로 측정되었다. 배양시간에 따른 HK-6의 세포외 단백질의 변화를 2-D PAGE 분석을 통하여 분석하였다. 그 결과 36시간배양 후에 가장 현저하게 유도된 spot #1을 분리하였으며, MALDI-TOF MS를 이용하여 단백질 동정을 실시한 결과, 유도된 단백질의 아미노산 서열은 $^1EKKIEKYREEEORLK^{15}$으로서, serine protein kinase (PrkA) (AAU22526)로 확인되었다.

• 한국미생물·생명공학회지
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• 제36권3호
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• pp.209-214
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• 2008

VBNC(Viable but nonculturable)란 생존에 불리한 환경하에서 살아 있으나 일반 영양배지에서 자라지 못하는 미생물의 상태를 나타낸다. 본 연구는 구리를 이용해 Escherichia coli에서 VBNC를 유도하고 이의 특성을 살펴보았다. 구리를 처리한 후 전통적인 평판 배양법에 의한 집락 형성계수(colony forming unit, CFU)를 측정한 결과 배양되지는 않으나, Live/Dead BacLight bacterial viability kit 염색 후 유세포계수기로 측정한 결과 살아있는 미생물로 계수되어 VBNC 상태가 확인하였다. VBNC 유도된 미생물로부터 genomic DNA와 RNA를 분리하고 이들의 안정성을 관찰하였는데 DNA에 비해 RNA의 붕괴가 많이 진행되었음을 확인할 수 있었고 RNA의 붕괴는 특정크기로 붕괴되는 것으로 관찰되었다. 또한 생물전용 투과전자현미경(Bio-Transmission Electron Microscope, Bio-TEM)을 통해 VBNC 세포의 형태를 관찰하였는데 VBNC 상태에서는 정상상태에 비해 periplasmic space가 온전하지 못하고 세포내막과 세포 외막이 분리되었으며 세포질의 양이 현저히 감소됨이 관찰되었다.

• 대한의생명과학회지
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• 제2권1호
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• pp.49-55
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• 1996

대장균군 검사용 간이 시험지는 본 실험실에서 국내 최초로 고안, 개발하였으며 이 간이 시험지법은 현장 검사법의 하나로 대장균군이 내는 succinic acid dehydrogenase 때문에 tetrazolium salt가 환원되어 적색 반점을 형성하는 것을 이용한 방법으로서 이 간이 시험지의 제조는 대체로 종래의 표준 평판법과 거의 동일한 조성의 배지와 시약을 사용하여 여지에 흡착시킨 후, 건조시켜($60^\circ$C) 멸균한 것으로 표준 평판법과 어떤 상관관계가 있는가를 검토하였다. 이 간이 시험지의 제조에서는 bile salt No. 3를 deoxycholate로 대체하여 제조 원가를 절감하였고, 또한 일본에서 현재 시판되고 있는 제품과 품질 비교시험을 하여 더 좋은 결과를 얻었으며 종래의 표준 평판법과 비교하였을 때도 오히려 표준 평판법 (24-48시간 배양)보다 빠른 시간(16-20시간 배양)내에 판정할 수 있는 이 점이 있으며, 표준 평판법에서는 없어서는 안될 배지나 배양 접시, pipette등의 자료 및 기구가 일체 필요없고 언제 어디서나 현장에서 직접 시험할 수 있어 매우 간편하며 또한 저렴한 가격으로 제조 할 수 있는 경제성이 높은 이점을 갖고 있다.

• 한국식품영양과학회지
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• 제38권12호
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• pp.1773-1778
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• 2009

본 연구는 인공합성보존료를 대체할 수 있는 천연보존료의 개발을 위한 방안으로 대표적인 세균성 식중독 미생물로서 그람 음성균인 Escherichia coli O157:H7과 Salmonella Typhimurium, 그리고 그람 양성균인 Staphylococcus aureus와 Listeria monocytogenes에 대한 항균 특성을 조사하였다. 구아바 가지와 잎의 메탄올과 에탄올 추출물 S. aureus의 생육을 저해하였으며, 아울러 가지와 잎의 아세톤 추출물은 200 ppm 농도의 첨가에서 식중독 원인균 S. aureus균을 전혀 자라지 못하는 결과를 확인하였다. 이러한 결과는 구아바 잎과 가지의 추출물에 그람 양성균에 대한 높은 항균활성이 있음을 의미하며, 구아바 가지를 항균력을 가지는 소재로 이용될 수 있음을 시사한다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.196-196
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• 2017

Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

• 대한치과보철학회지
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• 제44권3호
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• pp.284-294
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• 2006

Statement of problem. Proliferation of Candida albicans is primarily within the plaque on the fitting surface of the denture rather than on the inflamed mucosa. Consequently, the treatment of the denture is equally important as treatment of the tissue. Cleansing and disinfection should be efficiently carried-out as the organisms can penetrate into the voids of the acrylic resin and grow in them, from which they can continue to infect and reinfect bearing tissues. Purpose. The purpose of this study was to evaluate the applicability of photocatalytic reaction to eliminate Candida albicans from acrylic resin denture base, and to investigate the anti-fungal effect with various UVA illumination time. Materials and Methods. The specimens were cured by the conventional method following the manufacturer's instruction using thermal polymerized denture base resin (Vertex RS: Dentimex, Netherlands). $TiO_2$ photocatalyst sol(LT), which is able to be coated at normal temperature, was made from the Ti-alkoxide progenitor. The XRD patterns, TEM images and nitrogen absorption ability of the $TiO_2$ photocatalyst sol(LT) were compared with the commercial $TiO_2$ photocatalyst P-25. The experimental specimens were coated with the mixture of the $TiO_2$ photocatalyst sol(LT) and binder material (silane) using dip-coater, and uncoated resin plates were used as the control group. Crystallinity of $TiO_2$ of the specimen was tested by the XRD. Size, shape and chemical compositions were also analyzed using the FE-SEM/ EDS. The angle and methylene blue degradation efsciency were measured for evaluating the photocatalytic activity of the $TiO_2$ film. Finally, the antifungal activity of the specimen was tested. Candida albicans KCTC 7629(1 ml, initial concentration $10^5$ cells/ ml) were applied to the experiment and control group specimens and subsequently two UVA light source with 10W, 353 nm peak emission were illuminated to the specimens from 15cm above. The extracted $2{\mu}l$ of sample was plated on nutrient agar plate ($Bacto^{TM}$ Brain Heart Infusion; BD, USA) with 10 minute intervals for 120 minute, respectively. It was incubated for 24 hours at $37^{\circ}C$ and the colony forming units (CFUs) were then counted. Results. Compared the characteristics of LT photocatalyst with commercial P-25 photocatalyst, LT were shown higher activity than P-25. The LT coated experimental specimen surface had anatase crystal form, less than 20 nm of particle size and wide specific surface area. To evaluate the photocatalytic activity of specimens, methylene blue degradation reaction were used and about 5% of degradation rate were measured after 2 hours. The average contact angle was less than $20^{\circ}$ indicating that the LT photocatalyst had hydrophilicity. In the antifungal activity test for Candida albicans, 0% survival rate were measured within 30 minute after irradiation of UVA light. Conclusion. From the results reported above, it is concluded that the UVA-LT photocatalytic reaction have an antifungal effect on the denture surface Candida albicans, and so that could be applicable to the clinical use as a cleaning method.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2016년도 춘계학술대회 및 임시총회
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• pp.13-13
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• 2016

Insects constitute the largest and most diverse group of animals in the world. They also serve as the hosts or nutrient sources for an immense assemblage of pathogens, parasites, and predators. More than 700 fungal species from 100 genera have adopted an entomopathogenic lifestyle. Although entomopathogenic fungi were studied as only biocontrol agents against a variety of pests in various countries, it has been recently focused their additional roles in nature. They are antagonists to/against plant pathogens, endophytes, and possibly even plant growth promoting agents. The potential antimicrobial effect against fungal plant pathogens by an isolate of entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have been reported since late 1990s, but wasn't reported pathogenicity of the isolate against pests. Later, a Canadian Lecanicillium sp. isolate and L. longisporium isolated from Vertalec$^{(R)}$ showed simultaneous control effect against both aphid and cucumber powder mildew. Therefore, the antimicrobial activities of 342 fungi isolates collected from various regions and conditions in Korea were evaluated against plant pathogenic fungus Botrytis cinerea using dual culture technique on agar plate. As a result, 186 isolates (54.4%) shown the antifungal activity against B. cinerea. The culture filtrates of selected fungi completely suppressed the growth of the microorganisms, indicating that suppression was due to the presence of antimicrobial substances in the culture filtrate. Mode of action of these fungi against insect involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During infection process, secreted enzymes, proteinous toxins, and/or secondary metabolites secreted by entomopathogenic fungi can be used to overcome the host immune system, modify host behavior, and defend host resources. Recently, secondary metabolites isolated from entomopathogenic fungi have been reported as potential bioactive substances. Generally, most of bioactive substances produced by entomopathogenic fungi have reported low molecular weight (lower than 1,000 g/mol) as peptide and, in contrast the high molecular weight fungal bioactive substances are rare. Most substances based on entomopathogenic fungi were shown antimicrobial activity with narrow control ranges. In our study we analyzed the antimicrobial substances having antagonistic effects to B. cinerea. Antimicrobial substances in our fungal culture filtrates showed high thermostability, high stability to proteolytic enzymes, and hydrophilicity and their molecular weights were differed from substance. In conclusion, entomopathogenic fungi showed pathogenicity against insect pests and culture filtrate of the fungi also shown to antimicrobial activity. In the future, we can use the entomopathogenic fungi and its secondary metabolites to control both insect pest control and plant pathogenic fungi simultaneously.

• 한국산림과학회지
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• 제85권2호
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• pp.329-339
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• 1996

곰솔(Pinus thunbergii)유묘(幼苗)에 Pisolithus tinctorius(Pt., KJ-1)를 접종(接種)하고 $CuCl_2$용액을 0, 40, 120, 260, 430ppm 농도(濃度)로 처리수준(處理水準)을 달리하여 균근형성(菌根形成)및 발달(發達), 유묘생장(幼苗生長), 양분흡수(養分吸收)에 미치는 영향(影響)을 생장(生長) 시기별(時期別)(6월(月) 1일(日), 7월(月) 1일(日), 8월(月) 1일(日), 9월(月) 1일(日), 10월(月) 1일(日), 11월(月) 1일(日))로 조사(調査)하여 얻어진 결과(結果)는 다음과 같다. 1. 곰솔종자(種子)의 발아율(發芽率)은 균근접종구(菌根接種區)가 비접종구(非接種區)보다 높았다. 곰솔유묘(幼苗)의 균근형성율(菌根形成率)은 고농도(高濃度)로 갈수록 현저히 감소(減少)하였으며, Pt. 균(菌)의 Cu 내성조사(耐性調査)를 위해 실시한 MMN Agar평판배지실험(培地實驗)에서도 고농도(高濃度)로 갈수록 생장(生長)이 감소(減少)하였다. 2. 곰솔유묘(幼苗)의 묘고생장(苗高生長)에서 접종구(接種區) 40ppm구(區)와 120ppm구(區)에서 생장효과(生長效果)가 크게 나타났으며, 비접종구(非接種區) 430ppm구(區)에서 가장 저조(低調)했다. 3. 균근(菌根)의 외부형태(外部形態)는 0, 40ppm구(區)에서는 monopodial type, fern-like type, cluster-like type이 나타났으나, 260, 430ppm에서는 monopodial type만이 나타났다. 4. 접종구(接種區)에서 근(根)길이, 유기엽(幼期葉)의 수(數), 유기엽(幼期葉)의 총(總)길이, 총건중량(總乾重量), 본엽수(本葉數), 본엽총(本葉總)길이 등(等)이 비접종구(非接種區)보다 높게 나타났으며, 일반적(一般的)으로 접종구(接種區) 40, 120ppm구(區)에서 생장효과(生長效果)가 크게 나타났다. 5. 곰솔유묘(幼苗) 생육배지(生育培地)인 토양분석(土壤分析) 결과(結果)는 pH, Na, CEC는 고농도(高濃度)로 갈수록 증가(增加)하는 경향(傾向)을 나타내었고, 전질소(全窒素), 유기물(有機物), P, K, Ca, Mg는 40, 120ppm구(區)에서 감소(減少)하였다. 토양내(土壤內) Cu 분석결과(分析結果) 0ppm구(區)에서는 접종구(接種區)에서 높게 나타났으며, 고농도(高濃度)로 갈수록 뚜렷하게 비접종구(非接種區)에서 높은 함량(含量)이 나타났다. 6. 지상부(地上部) 분석결과(分析結果) 접종구(接種區)에서 전질소(全窒素), P, K의 흡수(吸收)가 많았고, 비접종구(非接種區)에서 Cu 흡수(吸收)가 많았다. 지하부(地下部) 분석결과(分析結果) 전질소(全窒素)는 생장(生長)이 좋은 40, 120ppm구(區)에서 많은 함량(含量)을 나타내었고, P은 비접종구(非接種區)보다 접종구(接種區)에서 아주 높은 흡수(吸收)가 있었고, Cu는 비접종구(非接種區)에서 높은 함량(含量)을 나타냈다. 그리고 모든 처리구(處理區)에서 지상부(地上部)보다 지하부(地下部)에서 현저(顯著)하게 높은 Cu 함량(含量)이 나타났다.