• Title/Summary/Keyword: Bacillus subtilis spore

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Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Dongchimi (동치미로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성)

  • 주길재;이창호;우철주
    • Journal of Life Science
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    • v.11 no.5
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    • pp.432-438
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    • 2001
  • Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

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Characterization of a Chitinase Gene Exhibiting Antifungal Activity from a Biocontrol Bacterium Bacillus licheniformis N1

  • Lee, Kwang-Youll;Heo, Kwang-Ryool;Choi, Ki-Hyuck;Kong, Hyun-Gi;Nam, Jae-Sung;Yi, Young-Byung;Park, Seung-Hwan;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.344-351
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    • 2009
  • A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

Inhibition of growth and toxin production of ochratoxigenic Aspergillus spp. by isolated bacteria (분리세균에 의한 ochratoxin 생성 Aspergillus spp.의 생장 및 독소생성 저해)

  • Hwang, Ji-Seon;Choi, Ho-Yeong;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.226-233
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    • 2019
  • Ochratoxin A (OTA) that is one of mycotoxins produced mainly by Aspergillus spp. is a common contaminant of stored grains and poses health hazards to human and livestock. The aim of this study is to explore the ability of isolated bacteria Bacillus subtilis AF13 and Streptomyces shenzhenensis YR226 to inhibit growth and OTA production of 3 ochratoxigenic Aspergillus strains. The antifungal activity against mycelial growth and sporulation of Aspergillus strains was examined by coculture with AF13 and YR226 on potato dextrose agar plate. AF13 and YR226 reduced 77.58 and 78.48% of fungal colony radius, respectively, and both strains inhibited fungal sporulation up to 99% in 10 days of incubation. YR226 also reduced more than 91% of spore germination of 3 fungal strains. When Aspergillus strains were cocultured with AF13 or YR226 in yeast extract sucrose medium, mycelial growth and OTA production decreased in all three fungal strains. In particular, AF13 completely inhibited the mycelial growth of A. alutaceus and inhibited its OTA production by 99%, and YR226 also reduced mycelial growth and toxin production up to 99%, respectively. Antimicrobial substances produced by AF13 and YR226 included siderophore, chitinase, protease, ${\beta}$-1,3-glucanase and biosurfactant. These results suggest that AF13 and YR226 can be used in a biological method to prevent valuable crops against mycotoxigenic fungi, and therefore decrease economic damage in agriculture and feed industry.

Desmutagenicity of Enzymatically Browned Substances Obtained from the Reaction of Prunus salicina (Red) Enzyme and Polyphenols (재래종 적색자두(Prunus salicina) 효소갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi;Hong, Eun-Hee;Omura, Hirohisa
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.212-219
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    • 1987
  • The rec-assay on Bacillus subtilis strains H17$({Rec}^+)$ and M45$({Rec}^-)$, the Ames test with modification of preincubation on Salmonella typhimurium TA98 and TA100 and DNA-breaking test on double strand calfthymus DNA were carried out using enzymatically browned substances obtained from the reaction of Prunus salicina (Red) enzyme and polyphenols. The spore rec-assay of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone. 3,4-dihydrohyoluene and chlorogenic acid showed non-mutagenic activity The spore rec-assay showed a little influence of ${Zn}^{2+}$ and ${Ni}^{2+}$ on the action of four kinds of enzymatic browning reaction products. The enzymatic browning reaction products of polyphenols did not show DNAbreaking activity. ${Cu}^{2+}$ of various metal ions influenced on DNA-breaking of enzymatic browning reaction products of pyrogallol. However, enzymatic browning reaction products of chlorogenic acid inhibited on DNA-breaking activity. Four kinds of enzymatic browning reaction products showed non-mutagenic activity on Salmonella typhimurium TA98 and TA100 with S-9 mix. In the mutagenicity on Salmonella typhimurium TA98 and TA100 with S-9 mix in the presence of benzo$({\alpha})$pyrene which is the carcinogenic substances, four kinds of enzymatic browning reaction products showed desmutagenic activity.

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Antimutagenic Effects of Browning Products Reacted with Polyphenol Oxidase Extracted from Apple (사과 Polyphenol Oxidase에 의한 효소갈변반응 생성물의 항돌연변이 효과)

  • Baik, Chang-Weon;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.625-631
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    • 1990
  • This study was carried out to investigate the antimutagenic effects of five kinds of apple enzymatic browning reaction products(AEBRP) on mitomycin C (MMC), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroquinoline-1-oxide(4NQO), benzo(${alpha}$)pyrene(B(${alpha}$)P) and 3-amino-1,4-dimethyl-5H-pyri-do [4,3-b]indol (Trp-P-1). In spore rec-assay using B. subtilis Hl7($rec^+$) and M45($rec^-$), homocatechol-AEBRP and hydroquinone-AEBRP showed strong antimutagenic effects on MMC and MNNG as the concentration of AEBRP increased. In the Ames test using S. typhimurium TA98 and TA100, hydroxyhydroquinone-AEBRP and pyrogallol-AEBRP showed strong antimutagenic effects on Trp-p-1 and B(${alpha}$)p in TA98 and TA100 in the presence of S-9 mix. Most of AEBRPs suppressed about 50% to 80% the mutagenesis in S. typhimurium TA98 induced by MNNG, however, AEBRPs except hydroxyhydroquinone-AEBRP showed antimutagenic effects of about 94% in TA100. Antimutagenic effects of the five kinds of AEBRPs on 4-NQO were more or less weak, in particular homocatechol-AEBRP exhibited the inhibitory effect of about 48% in TA98, and homocatechol-AEBRP and hydroquinone-AEBRP showed inhibitory effects of about 46% to 58% in TA 100.

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Survival of Microorganisms on Antimicrobial Filters and the Removal Efficiency of Bioaerosols in an Environmental Chamber

  • Kim, Sung Yeon;Kim, Misoon;Lee, Sunghee;Lee, JungEun;Ko, GwangPyo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1288-1295
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    • 2012
  • Exposure to bioaerosols causes various adverse health effects including infectious and respiratory diseases, and hypersensitivity. Controlling exposure to bioaerosols is important for disease control and prevention. In this study, we evaluated the efficacies of various functional filters coated with antimicrobial chemicals in deactivating representative microorganisms on filters or as bioaerosols. Tested functional filters were coated with different chemicals that included (i) Ginkgo and sumac, (ii) Ag-apatite and guanidine phosphate, (iii) $SiO_2$, ZnO, and $Al_2O_3$, and (iv) zeolite. To evaluate the filters, we used a model ventilation system (1) to evaluate the removal efficiency of bacteria (Escherichia coli and Legionella pneumophila), bacterial spores (Bacillus subtilis spore), and viruses (MS2 bacteriophage) on various functional filters, and (2) to characterize the removal efficiency of these bioaerosols. All experiments were performed at a constant temperature of $25^{\circ}C$ and humidity of 50%. Most bacteria (excluding B. subtilis) rapidly decreased on the functional filter. Therefore, we confirmed that functional filters have antimicrobial effects. Additionally, we evaluated the removal efficiency of various bioaerosols by these filters. We used a six-jet collision nebulizer to generate microbial aerosols and introduced it into the environmental chamber. We then measured the removal efficiency of functional filters with and without a medium-efficiency filter. Most bioaerosol concentrations did not significantly decrease by the functional filter only but decreased by a combination of functional and medium-efficiency filter. In conclusion, functional filters could facilitate biological removal of various bioaerosols, but physical removal of these by functional was minimal. Proper use of chemical-coated filter materials could reduce exposure to these agents.

Reduction Effect of Microorganisms by Nano Plasma ion (NPi) (Nano Plasma ion (NPi)에 의한 미생물 제어)

  • Kang, Hyeon-Cheol;Yun, Han-Seong;Sung, Bong-Jo;Lee, Sung-Hwa;Lee, Jang-Woo;Seo, Yong-Bae;Lee, Myung-Suk
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1710-1715
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    • 2011
  • The bactericidal effect of nano plasma ion (NPi) which was generated by NPi was analyzed using different kinds of microorganisms, exposure times, chamber sizes, ion amounts and distance. As the result of Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella pneumoniae, Staphylococcus aureus and Bacillus subtilis were shown different in decrement. Gram-negative bacteria E. coli showed the highest percentage (96.57%) and Gram-positive bacteria B. subtilis which produced spore has the lowest percentage (57.41%). From the exposure time of NPi most of the microorganisms were extinct at an early stage. According to the size of the chamber we compared the loss of E.coli and the experiment result shown, analyzed NPi using 5 chambers $0.005m^3$ to $30m^3$ for 2 hr, that when volume of the chamber increased, saturation ion and bactericidal effect was decreased. In addition, an NPi generator installed in the $1m^3$ chamber investigated the decrement of E. coli. Saturation ion concentration increased with decrement. Finally, E. coli showed a similar reduction according to the distance from NPi generator.

Screening for Effective Organic Farming Materials for the Control of Tuber Rot of Gastrodia elata Caused by Fusarium spp. (천마 괴경썩음병 방제에 효과적인 유기농업자재 선발)

  • Kim, Chang-Su;Lee, Han-Na;Kim, Hyo-Jin;Seo, Sang-Young;Jo, Jong-Hyun;Lee, Eun-Suk;Ahn, Min Sil;Kim, Hee-Jun
    • Korean Journal of Organic Agriculture
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    • v.29 no.4
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    • pp.561-574
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    • 2021
  • This study was conducted to evaluate the control efficacy of the organic farming materials (OFMs) on tuber rot of Gastrodia elata caused by Fusarium spp. The antifungal activities in vitro as well as the suppressive effect of 15 OFMs on the spore germination and germ tube growth by inoculating spore suspension on immature tubers in vivo were investigated. 7 OFMs inhibited the mycelial growth of Fusarium spp. and 7 of them were microbial agents. In the screening using immature tubers, 3 OFMs were very effective with control efficacy value of 70%. Among them, sulfur provided suppressive effect on both mycelial growth and spore germination against tuber rot of G. elata. Finally, 3 OFMs were selected to test the protective and curative effects, and all chosen OFMs significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, sulfur and Bacillus subtilis gave excellent protective control efficacy with control values of 93.2% and 86.9%, respectively, whereas its curative control effect was relatively low (73.3%, 60.2%). On the other hand, the preventive and curative effects of citronella + paraffin oil + ethyl alcohol were 73.3% and 67.0%, respectively. This study suggests that tuber rot of G. elata can be controlled by some OFMs in the rain shelter greenhouse under continuous cultivation condition and protective treatment is more important and efficient.

Characteristics of Microorganisms Contaminating Seafood Cooking Drips Exposed to Gamma Irradiation (감마선 조사된 수산 자숙액의 오염 미생물군 특성)

  • Choi, Jong-Il;Kim, Yeon-Joo;Kim, Jae-Hun;Chun, Byung-Soo;Ahn, Dong-Hyun;Kwon, Joong-Ho;Hwang, Young-Jung;Byun, Myung-Woo;Lee, Ju-Woon
    • Food Science and Preservation
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    • v.16 no.2
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    • pp.286-291
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    • 2009
  • Microorganisms in seafood cooking drips were counted and identified. Total viable cell counts were 6.40 and 3.10 log CFU/g in cooking drips of Hizikia fusiformis and Thunnus thynnus, respectively. However, microbial populations fell with increased irradiation doses. In H. fusiformis cooking drips, a 5-log reduction in total aerobic bacteria was obtained by irradiation with 5 kGy. In T. thynnus cooking drips, however, contaminating microorganisms were more resistant to gamma irradiation and only a 1-log reduction was seen. DNA sequence analysis showed that the principal contaminating microorganisms in H. fusiformis and T. thynnus cooking drips were Lactobacillus and Bacillus species, respectively. Therefore, the high irradiation resistance of T. thynnus cooking drips microbes may result from spore formation by Bacillus species.

The Overexpression of Subtilisin Enzyme Using Mutations on Transition State Regulatory Proteins of AprE Promoter and Development of Bacillus subtilis Host System (AprE Promoter전이상태 조절인자 변이주를 이용한 공업적 효소의 과발현과 고초균 숙주계의 개발)

  • 류성호;박승환김병기
    • KSBB Journal
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    • v.11 no.1
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    • pp.8-14
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    • 1996
  • Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

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