• Title/Summary/Keyword: cyt gene

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Distribution of Toxin Genes and Enterotoxins in Bacillus thuringiensis Isolated from Microbial Insecticide Products

  • Cho, Seung-Hak;Kang, Suk-Ho;Lee, Yea-Eun;Kim, Sung-Jo;Yoo, Young-Bin;Bak, Yeong-Seok;Kim, Jung-Beom
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2043-2048
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    • 2015
  • Bacillus thuringiensis microbial insecticide products have been applied worldwide. Although a few cases of B. thuringiensis foodborne illness have been reported, little is known about the toxigenic properties of B. thuringiensis isolates. The aims of this study were to estimate the pathogenic potential of B. thuringiensis selected from microbial insecticide products, based on its possession of toxin genes and production of enterotoxins. Fifty-two B. thuringiensis strains selected from four kinds of microbial insecticide products were analyzed. PCR assay for detection of toxin genes and immunoassay for detection of enterotoxins were performed. The hemolysin BL complex as a major enterotoxin was produced by 17 (32.7%), whereas the non-hemolytic enterotoxin complex was detected in 1 (1.9%) of 52 B. thuringiensis strains. However, cytK, entFM, and ces genes were not detected in any of the tested B. thuringiensis strains. The potential risk of food poisoning by B. thuringiensis along with concerns over B. thuringiensis microbial insecticide products has gained attention recently. Thus, microbial insecticide products based on B. thuringiensis should be carefully controlled.

Manufacturing Protein-DNA Chip for Depigmenting Agent Screening (전사인자 저해제 통한 미백제 탐색용 단백질 칩 제작)

  • Han Jung-Sun;Kwak Eun-Young;Lee Hyang-Bok;Shin Jlung-Hyun;Baek Seung-Hak;Chung Bong-Hyun;Kim Eun-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.479-483
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    • 2004
  • An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.

Mitochondrial Efficiency-Dependent Viability of Saccharomyces cerevisiae Mutants Carrying Individual Electron Transport Chain Component Deletions

  • Kwon, Young-Yon;Choi, Kyung-Mi;Cho, ChangYeon;Lee, Cheol-Koo
    • Molecules and Cells
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    • v.38 no.12
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    • pp.1054-1063
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    • 2015
  • Mitochondria play a crucial role in eukaryotic cells; the mitochondrial electron transport chain (ETC) generates adenosine triphosphate (ATP), which serves as an energy source for numerous critical cellular activities. However, the ETC also generates deleterious reactive oxygen species (ROS) as a natural byproduct of oxidative phosphorylation. ROS are considered the major cause of aging because they damage proteins, lipids, and DNA by oxidation. We analyzed the chronological life span, growth phenotype, mitochondrial membrane potential (MMP), and intracellular ATP and mitochondrial superoxide levels of 33 single ETC component-deleted strains during the chronological aging process. Among the ETC mutant strains, 14 ($sdh1{\Delta}$, $sdh2{\Delta}$, $sdh4{\Delta}$, $cor1{\Delta}$, $cyt1{\Delta}$, $qcr7{\Delta}$, $qcr8{\Delta}$, $rip1{\Delta}$, $cox6{\Delta}$, $cox7{\Delta}$, $cox9{\Delta}$, $atp4{\Delta}$, $atp7{\Delta}$, and $atp17{\Delta}$) showed a significantly shorter life span. The deleted genes encode important elements of the ETC components succinate dehydrogenase (complex II) and cytochrome c oxidase (complex IV), and some of the deletions lead to structural instability of the membrane-$F_1F_0$-ATP synthase due to mutations in the stator stalk (complex V). These short-lived strains generated higher superoxide levels and produced lower ATP levels without alteration of MMP. In summary, ETC mutations decreased the life span of yeast due to impaired mitochondrial efficiency.

Prevalence of Microbiological Contamination in the Ready-To-Eat Side Dishes Sold in Gyeongsangnam-do, South Korea (경남지역에서 유통되는 즉석 반찬류의 미생물 오염도 조사)

  • Ji-Yeon Um;Hye-Jeong Jang;Yeon-Ju Choi;So-Young Kim;Areum Jo;Min Young Kim;Jihee Ahn;Jea-Dong Kim
    • Journal of Food Hygiene and Safety
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    • v.38 no.4
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    • pp.217-227
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    • 2023
  • The consumption of ready-to-eat side dishes is rapidly growing in South Korea. These foods are particularly vulnerable to microbiological contamination as they are often cooked without any treatment, such as heating or stored at room temperature after cooking. Hence, in 2022, we analyzed the ready-to-eat side dishes sold in Gyeongsangnam-do, South Korea for microbiological contamination. We collected 100 samples from supermarkets in 7 cities, and then examined them for presence of food-borne pathogens and sanitary indicator bacteria. In the analysis of the food-borne pathogens, Bacillus cereus and Clostridium perfringens were isolated from 51 samples (51.0%) and 3 samples (3.0%), respectively. However, both quantitatively met the Korean Food Standards Codex. Genes of five different enterotoxins and one emetic toxin were analyzed from the 51 isolated B. cereus strains. We detected enterotoxin entFM (100.0%), nheA (94.1%), hblC (58.8%), cytK (56.9%), and bceT (41.2%) in 51 isolates, and emetic toxin gene, CER, in only one (2.0%) isolate. We did not detect C. perfringens toxin gene (cpe) that causes food poisoning in any one of the three C. perfringens isolates. In the case of sanitary indicator bacteria, Kimchi had the highest levels of total aerobic bacteria and coliforms, followed by Saengchae, Jeotgal, Jeolim, Namul, and Jorim, respectively. We counted total aerobic bacteria at two different storage temperatures (4℃ and 20℃) to determine the effect of storage temperature. When stored at 20℃, total aerobic bacteria count increased in most of the ready-to-eat side dishes, except for Jeotgal. This result conclusively shows the need for refrigerating the ready-to-eat side dishes after purchase. Further research is needed to assess the risk and safety of the ready-to-eat side dishes available in the market and determine appropriate safety management practices.

Analysis of Foodborne Pathogens in Food and Environmental Samples from Foodservice Establishments at Schools in Gyeonggi Province (경기지역 학교 단체급식소 식품 및 환경 중 식중독균 분석)

  • Oh, Tae Young;Baek, Seung-Youb;Koo, Minseon;Lee, Jong-Kyung;Kim, Seung Min;Park, Kyung-Min;Hwang, Daekeun;Kim, Hyun Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1895-1904
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    • 2015
  • Foodborne illness associated with food service establishments is an important food safety issue in Korea. In this study, foodborne pathogens (Bacillus cereus, Clostridium perfringens, Escherichia coli, pathogenic Escherichia coli, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Vibrio parahaemolyticus) and hygiene indicator organisms [total viable cell counts (TVC), coliforms] were analyzed for food and environmental samples from foodservice establishments at schools in Gyeonggi province. Virulence factors and antimicrobial resistance of detected foodborne pathogens were also characterized. A total of 179 samples, including food (n=66), utensil (n=68), and environmental samples (n=45), were collected from eight food service establishments at schools in Gyeonggi province. Average contamination levels of TVC for foods (including raw materials) and environmental samples were 4.7 and 4.0 log CFU/g, respectively. Average contamination levels of coliforms were 2.7 and 4.0 log CFU/g for foods and environmental swab samples, respectively. B. cereus contamination was detected in food samples with an average of 2.1 log CFU/g. E. coli was detected only in raw materials, and S. aureus was positive in raw materials as well as environmental swab samples. Other foodborne pathogens were not detected in all samples. The entire B. cereus isolates possessed at least one of the diarrheal toxin genes (hblACD, nheABC, entFM, and cytK enterotoxin gene). However, ces gene encoding emetic toxin was not detected in B. cereus isolates. S. aureus isolates (n=16) contained at least one or more of the tested enterotoxin genes, except for tst gene. For E. coli and S. aureus, 92.7% and 37.5% of the isolates were susceptible against 16 and 19 antimicrobials, respectively. The analyzed microbial hazards could provide useful information for quantitative microbial risk assessment and food safety management system to control foodborne illness outbreaks in food service establishments.

Prevalence and Toxin Characteristics of Bacillus cereus Isolated from Vegetables in Gwangju Metropolitan City (광주지역 유통 채소류의 Bacillus cereus 오염실태 및 독소 특성)

  • Cho, Sun-Ju;Jeong, So-Hyang;Seo, Yu-Jin;Kim, Tae-Sun;Lee, Hyang-Hee;Lee, Min-Gyou;Seo, Jung-Mi;Cho, Bae-Sik;Kim, Joung-Beom
    • The Korean Journal of Food And Nutrition
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    • v.33 no.2
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    • pp.142-148
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    • 2020
  • The purpose of this study was to investigate the prevalence, toxin gene profiles, and enterotoxin producing ability of Bacillus cereus isolated from environment-friendly vegetables and good agricultural practices (GAP) vegetables. A total of 49 vegetables including 40 environment-friendly vegetables and 9 GAP vegetables were tested. The Vitek 2 system was used to identify B. cereus and the PCR was used to detect 6 toxin genes, respectively. B. cereus was detected in 34 (69.3%) of 49 vegetables and the prevalence of B. cereus in GAP vegetables (44.4%) was lower than in the environment-friendly vegetables (75.0%). The detection rates of entFM, nheA, hblC, and cytK enterotoxin genes, respectively, among all isolates were 100%, 97.0%, 88.2%, and 73.5%, respectively. All of the isolates had at least one or more enterotoxin gene and 20 isolates (58.8%) had hemolysin BL enterotoxin producing ability. The risk of food poisoning from the environment-friendly vegetables and the GAP vegetables has been shown as constant. Thus, it is necessary to expand the supply of GAP vegetables showing lower B. cereus contamination than the environment-friendly vegetables. The characteristics of the environment-friendly vegetables and the GAP vegetables that must be consumed after cleaning should be disseminated to consumers regarding food poisoning prevention.

Prevalence and Toxin Characteristics of Microorganism on Hand Towels Using for Children in Child Care Center (보육시설 유아 사용 수건의 미생물 분포 및 독소 특성)

  • Kim, Jung-Beom;Kim, Nan-Yong;Kang, Suk-Ho;Do, Young-Sook;Eom, Mi-Na;Yoon, Mi-Hye;Lee, Jong-Bok
    • Journal of Food Hygiene and Safety
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    • v.28 no.2
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    • pp.138-145
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    • 2013
  • This study was conducted to evaluate the microbiological contamination on commonly used hand towels in the child care centers and to investigate the toxin gene and toxin production ability of food-borne pathogens. A total of 22 commonly used hand towels including 7 for before use and 15 for during use were tested. The average number of total aerobic bacteria and fungi were 6.2 log CFU/100 $cm^2$ and 4.1 log CFU/100 $cm^2$. Coliform bacteria were detected in 4 out of 7 before used towels (57.1%) and all of during used towels (100%). These results showed that the sanitary conditions of hand towels in the child care centers should be improved promptly. Among the pathogenic bacteria, Staph. aureus and B. cereus without Salmonella spp. were detected in 5 (22.7%) and 11 (50.0%) out of 22 hand towels. All of Staphy. aureus isolated in this study did not possess any toxin genes and did not produce enterotoxin. The detection rate of hblC, hblD, and hblA toxin genes in B. cereus was 72.7, 72.7, and 54.5% respectively. The possession rate of nheA, nheB, and nheC toxin genes showed 81.8, 72.7, and 54.5% respectively. The cytK and entFM toxin genes were presented at 45.5 and 90.0% in B. cereus. The HBL was detected in 8 out of 11 B. cereus isolates (72.7%) and 5 B. cereus isolates produced NHE (45.5%). Ten out of eleven B. cereus isolates (90.9%) produced one or more enterotoxin such as HBL and NHE. From the results, using a private hand towel or paper towel is required to prevent the cross-contamination between commonly used hand towel and children's hands in the child care center.

Plumbagin from Plumbago Zeylanica L Induces Apoptosis in Human Non-small Cell Lung Cancer Cell Lines through NF-κB Inactivation

  • Xu, Tong-Peng;Shen, Hua;Liu, Ling-Xiang;Shu, Yong-Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2325-2331
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    • 2013
  • Objective: To detect effects of plumbagin on proliferation and apoptosis in non-small cell lung cancer cell lines, and investigate the underlying mechanisms. Materials and Methods: Human non-small cell lung cancer cell lines A549, H292 and H460 were treated with various concentrations of plumbagin. Cell proliferation rates was determined using both cell counting kit-8 (CCK-8) and clonogenic assays. Apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry and TUNEL assay. The levels of reactive oxygen species (ROS) were detected by flow cytometry. Activity of NF-${\kappa}B$ was examined by electrophoretic mobility shift assay (EMSA) and luciferase reporter assay. Western blotting was used to assess the expression of both NF-${\kappa}B$ regulated apoptotic-related gene and activation of p65 and $I{\kappa}B{\kappa}$. Results: Plumbagin dose-dependently inhibited proliferation of the lung cancer cells. The IC50 values of plumbagin in A549, H292, and H460 cells were 10.3 ${\mu}mol/L$, 7.3 ${\mu}mol/L$, and 6.1 ${\mu}mol/L$ for 12 hours, respectively. The compound concentration-dependently induced apoptosis of the three cell lines. Treatment with plumbagin increased the intracellular level of ROS, and inhibited the activation of NK-${\kappa}B$. In addition to inhibition of NF-${\kappa}B$/p65 nuclear translocation, the compound also suppressed the degradation of $I{\kappa}B{\kappa}$. ROS scavenger NAC highly reversed the effect of plumbagin on apoptosis and inactivation of NK-${\kappa}B$ in H460 cell line. Treatment with plumbagin also increased the activity of caspase-9 and caspase-3, downregulated the expression of Bcl-2, upregulated the expression of Bax, Bak, and CytC. Conclusions: Plumbagin inhibits cell growth and induces apoptosis in human lung cancer cells through an NF-${\kappa}B$-regulated mitochondrial-mediated pathway, involving activation of ROS.

Molecular Characterization and Toxin Profile of Bacillus cereus Strains Isolated from Ready-to-eat Foods (유통 중인 즉석·편의식품류에서 분리한 Bacillus cereus의 산생 Toxin 및 분자유전학적 특성 조사)

  • Kim, Tae Sun;Kim, Min Ji;Kang, Yu Mi;Oh, Geune;Choi, Su Yeon;Oh, Mu Sul;Yang, Yong Shik;Seo, Jung-Mi;Ryu, Mi-Geum;Kim, Eun-Sun;Ha, Dong-Ryong;Cho, Bae Sik
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.334-340
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    • 2014
  • Toxin-producing Bacillus cereus is the causative agent of two different types of food poisoning: the emetic and the diarrheal types. This study was conducted to investigate the presence of enterotoxin and emetic toxin genes in 263 B. cereus isolated from 619 different ready-to-eat food items. Hemolytic enterotoxins hblA, hblC, and hblD were detected in 85.6, 41.1, and 76.8%, respectively, of the B. cereus isolates. About 67.0% (175/263) of the isolates presented all of three genes. Non-hemolytic enterotoxins nheA, nheB, and nheC were detected in 100, 97.0, and 68.4% of the isolates, respectively. Approximately 90.0% (236/263) of the isolates presented all of these three non-hemolytic enterotoxin genes. Emetic toxin gene, CER, was detected in 132 of 263 (50.2%) isolates. Computer-assisted cluster analysis of Rep-PCR profiles showed a high genetic diversity among the isolates. All B. cereus isolates from food samples tested in this study carried at least 6 of 10 toxin genes.