Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus

홍성욱,Kim Jong-Hui,Cha Hyun A,정건섭,Bae Hyo Ju,Park Won Seo,Ham Jun-Sang,Park Beom-Young,Oh Mi-Hwa 한국미생물·생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.11

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0–70°C and pH 2–10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.

자궁경부암에서 종양진행과 E-cadherin/α-catenin 발현이상에 관한 연구

김정식,허진숙,김홍배,심정원,전현아,이근영,강성원 대한부인종양 콜포스코피학회 2003 Journal of Gynecologic Oncology Vol.14 No.2

목적 : E-cadherins은 상피특이적이고 칼슘 의존적 세포간 부착에 관여하는 막간 당단백질군에 속한다. E-cadherin의 기능은 그것과 세포질에서 연결되는 α-catenin에 의해 조절된다. α-catenin은 actin과 E-cadherin을 연결시킨다. 이 연구는 E-cadherin a/α-catenin의 발현이 양성자궁질환, 상피내암, 침윤암에서 어떤 차이를 보이는지 알아보는 것이다. 연구 방법 : 89개의 조직표본을 펀치생검, 원추 절제술, 자궁적출술에 의해 얻었다. 절편은 FIGO staging에 따라 양성 경부, 상피내암, 침윤암으로 분류되었다. E-cadherin/α-catenin의 발현은 monoclonal antibody로 면역조직화학 염색을 하여 조사하였다. 세 군간의 E-cadherin/α-catenin의 발현양상을 조사했다. 통계적 의미를 조사하기 위해 x² test를 이용했다. 결과 : E-cadherin에서는 정상염색(normal), 낮은 양성(low-positive), 이종(異種, heterogeneous)염색, 음성(negative) 염색 결과가 양성 질환인 경우 각각 19, 1, 0, 2예, 상피내암인 경우 12, 5, 3, 4예, 침윤암인 경우 10, 8, 20, 5예를보였다. α-catenin에서도 양성 질환인 경우 각각 18, 1, 1, 2예, 상피내암인 경우 9, 9, 4, 2예, 침윤암인 경우 8, 8, 19, 8예를 보였다. 조직검사와 E-cadherin과 α-catenin의 발현강도와 분포사이에 통계적으로 의미있는 상관관계 (χ² test, p<0.0001)를 보였다. 즉 양성질환 보다 상피내암, 침윤암일 수록 두 물질의 비정상적인 발현양상을 관찰할 수 있었고, E-cadherin/α-catenin의 발현도 변화와 조직학적 진단간에 상관관계가 있었다. 결론 : 이 결과에 따라 저자는 자궁경부암에서 종양진행에 따라 E-cadherin/α-catenin의 발현에 있어서 분포와 강도가 변화함을 알게 되었다. Objective : E-cadherins are a family of transmembrane glycoproteins involved in epithelial specific and calcium-dependent intercellular adhesion. E-cadherin function is regulated by its associated cytoplasmic protein, α-catenin, which links E-cadherin to the actin-based cytoskeleton. This study is to determine what lack and differrence in expression of E-cadherin and α-catenin between benign, CIS, and invasive carcinoma of uterine cervix. Methods : 89 tissue specimens were obtained by punch biopsy, conization, and hysterectomies. Sections were classified according to FIGO staging as benign cervix, carcinoma in situ, invasive cervical carcinoma. The expression of E-cadherin and α-catenin was examined immunohistochemically with monoclonal antibodies. What degree in expression of E-cadherin and α-catenin in three group was investigated. The χ² test was used to assess the statistical significance of this study. Results : The correlation between the altered expression of E-cadherin/α-catenin and the histopathologic diagnosis was statistically significant. Conclusion : According to this result, we found that the distribution and intensity in expression of E-cadherin and α-catenin is altered during tumor progression of cervical carcinoma.

Correlation between Attractions and Susceptibilities of Rice Varieties to Aphelenchoides besseyi Christie, 1942

Young-Bae Lee,A. A. F. Evans 한국응용곤충학회 1973 한국응용곤충학회지 Vol.12 No.4

벼 이삭선충(심고선충), Aphelenchoides besseyi Christie에 대한 벼 품종의 유인성과 감수성과의 상관관계를 알기 위하여 실험실관찰과 온실내의 포트 시험을 수행하였다. 본 선충에 대한 벼묘추출액의 유인성과 선충의 증식률은 벼의 품종에 따라서 크게 차이가 났으며 이들 두 요인간에는 큰 상관관계를 나타내었다. 즉, 벼묘의 추출액이 선충에 대하여 강한 유인성을 나타낸 품종일수록 포장에서의 선충증식이 잘 되어 감수성이 높았다. A laboratory observation and a greenhouse pot experiment were carried out to know the correlationship between the attraction and the susceptibility of rice varieties to Aphelenchoides besseyi. Degrees of attraction of rice seedling extract to A. besseyi and the multiplication rates of the nematode varied greatly according to the rice varieties used. And there was strong correlation between the two variables, i.e., the more the seedling extract of a variety attracted by the nematode in agar observation, the more the variety was susceptible in greenhouse pot experiment.

Mannosylerythritol lipids ameliorate ultraviolet A-induced aquaporin-3 downregulation by suppressing c-Jun N-terminal kinase phosphorylation in cultured human keratinocytes

Bae, Il-Hong,Lee, Sung Hoon,Oh, Soojung,Choi, Hyeongwon,Marinho, Paulo A.,Yoo, Jae Won,Ko, Jae Young,Lee, Eun-Soo,Lee, Tae Ryong,Lee, Chang Seok,Kim, Dae-Yong The Korean Society of Pharmacology 2019 The Korean Journal of Physiology & Pharmacology Vol.23 No.2

Mannosylerythritol lipids (MELs) are glycolipids and have several pharmacological efficacies. MELs also show skin-moisturizing efficacy through a yet-unknown underlying mechanism. Aquaporin-3 (AQP3) is a membrane protein that contributes to the water homeostasis of the epidermis, and decreased AQP3 expression following ultraviolet (UV)-irradiation of the skin is associated with reduced skin moisture. No previous study has examined whether the skin-moisturizing effect of MELs might act through the modulation of AQP3 expression. Here, we report for the first time that MELs ameliorate the UVA-induced downregulation of AQP3 in cultured human epidermal keratinocytes (HaCaT keratinocytes). Our results revealed that UVA irradiation decreases AQP3 expression at the protein and messenger RNA (mRNA) levels, but that MEL treatment significantly ameliorated these effects. Our mitogen-activated protein kinase inhibitor analysis revealed that phosphorylation of c-Jun N-terminal kinase (JNK), but not extracellular signal-regulated kinase or p38, mediates UVA-induced AQP3 downregulation, and that MEL treatment significantly suppressed the UVA-induced phosphorylation of JNK. To explore a possible mechanism, we tested whether MELs could regulate the expression of peroxidase proliferator-activated receptor gamma ($PPAR-{\gamma}$), which acts as a potent transcription factor for AQP3 expression. Interestingly, UVA irradiation significantly inhibited the mRNA expression of $PPAR-{\gamma}$ in HaCaT keratinocytes, whereas a JNK inhibitor and MELs significantly rescued this effect. Taken together, these findings suggest that MELs ameliorate UVA-induced AQP3 downregulation in HaCaT keratinocytes by suppressing JNK activation to block the decrease of $PPAR-{\gamma}$. Collectively, our findings suggest that MELs can be used as a potential ingredient that modulates AQP3 expression to improve skin moisturization following UVA irradiation-induced damage.

Correlation between Attractions and Susceptibilities of Rice Varieties to Aphelenchoides besseyi Christie, 1942

Lee Young-Bae,Evans A.A.F. Korean Society of Applied Entomology 1973 한국식물보호학회지 Vol.12 No.4

벼 이삭선충(심고선충), Aphelenchoides besseyi Christie에 대한 벼 품종의 유인성과 감수성과의 상관관계를 알기 위하여 실험실관찰과 온실내의 포트 시험을 수행하였다. 본 선충에 대한 벼묘추출액의 유인성과 선충의 증식률은 벼의 품종에 따라서 크게 차이가 났으며 이들 두 요인간에는 큰 상관관계를 나타내었다. 즉, 벼묘의 추출액이 선충에 대하여 강한 유인성을 나타낸 품종일수록 포장에서의 선충증식이 잘 되어 감수성이 높았다. A laboratory observation and a greenhouse pot experiment were carried out to know the correlationship between the attraction and the susceptibility of rice varieties to Aphelenchoides besseyi. Degrees of attraction of rice seedling extract to A. besseyi and the multiplication rates of the nematode varied greatly according to the rice varieties used. And there was strong correlation between the two variables; i.e., the more the seedling extract of a variety attracted by the nematode in agar observation, the more the variety was susceptible in greenhouse pot experiment.

Bacillus firmus Cyclodextrin Glycosyltransferase의 정제 및 특성

손천배(Cheon-Bae Sohn),김성애(Seong-Ai Kim),박영아(Young-A Park),김명희(Myung-Hee Kim),문숙경(Sook-Kyung Moon),장순애(Sun-Ae Jang),이명선(Myung-Sun Lee) 한국식품영양과학회 1997 한국식품영양과학회지 Vol.26 No.2

Bacillus firmus가 생산하는 cyclodextrin glycosyltran-sferase(CGTase)를 ammonium sulfate 침전, DEAE-Sephadex A-50 및 Sephdex G-100 column chromatography로 분리, 정제하였다. 이 방법으로 수율은 12%, 정제도는 49배인 SDS-PAGE 상에서 단일 band의 효소 단백질을 얻을 수 있었다. 정제된 CGTase는 Phast system의 SDS-polyacrylamide gel 전기영동에 의하여 분자량은 80,000dalton, isoelectric focusing으로 등전점은 9.6으로 추정되는 단백질이었다. 이 효소의 최적 활성 pH와 온도는 8.0,65℃이였으며, pH와 열안정성은 pH 5.5~9.0,50℃이였다. Soluble starch를 기질로 하여 24시간 효소반응한 액의 α- : β- : γ-cyclodextrin의 생성비율은 0.01 : 2.90 : 1.00으로 β- 및 γ-cyclodextrin을 주로 생산하였다. The cyclodextrin glycosyltransferase(EC 3.2.1.19) from Bacillus firmus was purified by precipitating with ammonium sulfate followed by, DEAE-Sephadex A-50 column chromatography and Sephadex G-100 column chromatography. In this way, we were able to obtain the single band protein on SDS-PAGE with a yield of 12%, whose purity was 49 fold. The purified CGTase was identified as a protein having molecular weight of approximately 80,000 dalton and isoelectric point of 9.6. The optimum pH and temperature for the enzyme activity were 8.0 and 65℃, respectively. The enzyme was stable at between pH 5.5 and 9.0 and up to 50℃. After 24hr of enzyme reaction using soluble starch as substrate, the ratio of α-, β- and γ-cyclodextrin production was 0.01 : 2.90 : 1.00, respectively. And this CGTase pro-duced mainly β- and γ-cyclodextrin.

항균활성 천연물질을 이용한 반추위 메탄저감용 친환경 첨가제 개발

이아름,양진호,조상범,나종삼,심관섭,김영훈,배귀석,장문백,최빛나,신수진,최낙진,Lee, A-Leum,Yang, Jinho,Cho, Sang-Buem,Na, Chong-Sam,Shim, Kwan-Seob,Kim, Young-Hoon,Bae, Gui-Seck,Chang, Moon-Baek,Choi, Bitna,Shin, Su-Jin,Choi, Nag-Jin 한국유기농업학회 2014 韓國有機農業學會誌 Vol.22 No.3

본 연구는 천연물질에서 유래한 반추위 메탄저감용 친환경 첨가제 개발을 위해 각기 다른 종균을 이용하여 발효한 세신 추출물의 항균활성, 항산화활성 및 in vitro 반추위 발효시험을 체계적으로 실시하였다. 접종된 균주들의 성장효율을 알아보기 위해 실시한 생균수측정 결과 L. curvatus NJ40 균주에서 유의적으로 높은 균주성장을 나타냈다(p<0.05). 항균활성측정 결과는 대조구 대비 L. curvatus NJ40 및 L. plantarum NJ45 균주와 발효된 세신 추출물이 병원균에 대한 항균효과를 나타내는 것으로 조사되었다. In vitro 반추위 발효실험에 세신 및 발효 세신 추출물을 적용한 결과, 휘발성 지방산 생성량 대비 반추위 메탄 저감효과가 나타났다. 특히 반추위 미생물 활력 및 사료이용 효율을 대표할 수 있는 휘발성 지방산 생성효율에 대한 부정적 효과 없이, 오히려 휘발성지방산 생성효율을 향상시키면서 반추위 메탄 저감효과를 가져올 수 있는 것으로 나타났다. The present study was conducted to investigate effective starter culture to improve biological activity of Asarum sieboldii. Antibacterial activity, antioxidant activity and reduction of enteric rumen methane production were used as criterions for biological activity. Ground A. sieboldii was added in MRS broth at 10% (w/v) and fermented by different starter cultures. Weissella confusa NJ28, Weissella cibaria NJ33, Lactobacillus curvatus NJ40, Lactobacillus brevis NJ42, Lactobacillus plantarum NJ45 and Lactobacillus sakei NJ48 were used for starter culture strains. Each starter culture was inoculated with 1% (v/v) ratio and fermentation was performed at $30^{\circ}C$ with agitation (150 rpm) for 48 h. MRS broth for the control was employed without starter culture. Then the fermentation growth was dried and extracted using ethyl alcohol. The growth of starter culture was detected at NJ40, NJ42, NJ45 and NJ48. And the highest cell growth was found in NJ40. Antibacterial activity against to Staphylococcus aureus, Listeria monocytogens, Mannheimia haemolytica and Salmonella gallinarum were observed in the extract fermented by NJ40 and NJ45. All treatments showed antioxidant activities, however, there were no significant differences (p>0.05). In in vitro rumen fermentation, negative control (NC) and positive control (PC) were assigned to without extract and with non-fermented A. sieboldii extract. Significant suppression of gas productions were detected in positive control and treatments compared to negative control (p<0.05). However, total volatile fatty acid production was not suppressed. Significant methane reduction per total volatile fatty acid productions were found in positive control and NJ45 treatment (p<0.05). The present study suggested a fermentation of A. sieboldii using NJ45 strain could improve its biological activity and make possible for its use in bio additive for enteric rumen methane mitigation without suppression of animal productivity.

Assessment of Alcohol Level in Inebriated Victims of Sexual Assault using Ethyl Glucuronide

Young?Hoon Jo,Mia Kwon,Joon-Bae Lee,Jeong A Ko 한국분석과학회 2022 학술대회논문집 Vol.2022 No.5

식품 기준·규격의 합리적 관리 ·운영에 관한 연구(Ⅰ) : 두부,식용유지, 전분의 분말상 원료에 관하여 Powdered marerials of Tofu, Edible oil, Starch

김희연,홍진환,박혜경,한상배,박종석,이은주,이정성,송경희,최은희,최영준,소경아,성영제,이주엽 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

본 연구는 분말상 원료(전분, 대두분)의 저장유통시 품질변화를 건전성과 안전성 측면에서 검토하여 식품의 기준 ·규격 설정f.」 합지적으로 반영하고자 하였다. 전분과 패두분의 이물을 검사하고, 2종 대두분(중국산, 미국산)을 ?0일간 IS'c와 30t에 저장하면서 성상, 수분, 산가, 과산화물가, 지방산 조성,아플라톡신 BB의 생성여부를 분석하였다. 중국산 대두분 30'』 저장군에서 50일 이후 곰팡이가 발생한 것 이외에 모든 시료에서 양호한 성상을 나타내었고, 전분과 대두닥 모든 시료에서 이물은 검출되지 않았다. 산가는 중국산과 미국산 시료간에 편차를 보여 증국산 30"C, 50일 저장시료에서 약 10배끙도 증가하였고, 과산화물가는 저장 90일후 15'E 저장군은 약 2.3배, 30'c 저장군은 3배정도 증가하였다. 대두분의 주요 지방산은 linoleic acidf18. 2), oleic acid(18 : 1), palmitic acid(16 : 0)이었으며, 3개월 저장에 따른 지방산 함량의 변화는 미미하였다. 아플라톡신 B₁은 모든 시료에서 검출되지 않았다. The objective of this study was to investigate auality changes of type raw materials(starch, soy flour) during various storage conditions. Starchflour(source of China, U.S.A) were slfred under two temperature(15'C, 30'c) forSensory evaluation, (o.reign material test, moisture content, acid value, peroxide ualue,composition, aflatorin Bi were analyzed. China soy flour(stor·ed 30'f) was contaminated by fungj on 50 days. Except for soy flour contardnated by fungi, sensory characteristics were not changed and foreign mater;als were notdetected. Acid value of China soy flour(stored at 30'C, 90 days) was about 10 times higherlevel before storage and acid value was more influenced by source(China, USA). Peroxide valuewas in proportion with the storage period slightly. The major fatty acid of soy flour waslinoleic acid(18 . 2), an(3 the change of fatty acid composition was not observed. Aflatoxlrl Blwas not detected.

만성 승모판 패쇄부전증 환자에서의 안지오텐신 전환효소 억제제의 장기 투여 효과

김대희,이명묵,이해영,조현재,박승정,서재빈,서정원,양한모,윤창환,조상호,이준희,김용진,김명아,손대원,오병희,박영배 대한순환기학회 2004 Korean Circulation Journal Vol.34 No.2