• Title/Summary/Keyword: B.subtilis

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The Effect of Container Types on the Growth of Bacteria during Kimchi Fermentation (김치 발효 시 용기의 종류가 세균 생장에 미치는 영향)

  • Han, Kook-Il;Kim, Mi-Jung;Kwon, Hyun-Jung;Kim, Yong Hyun;Kim, Wan-Jong;Han, Man-Deuk
    • The Korean Journal of Food And Nutrition
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    • v.26 no.2
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    • pp.249-257
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    • 2013
  • This study is being performed to confirm the container effects during the fermentation processes of kimchi. Kimchi fermentation was prepared in the laboratory with four different types of containers; namely, a traditional Onggi vessel (Korean traditional clay pot, TOV), plastic airtight covered Onggi vessel (PAOV), plastic covered vessel (PCV) and plastic airtight covered vessel (PACV). The kimchi fermentation in the different containers was followed by taking samples at 48 hour intervals for 10 days. In all fermentation containers, the pH changes of kimchi were decreased with fermentation days, while salt content was the same for all types of containers. The number of lactic acid bacteria in kimchi were $1.09{\times}10^8$ $CFU/m{\ell}$ at first. But the TOV, PAOV, PCV, and PACV after fermentation for 10 days were $1.42{\times}10^{10}$, $9.13{\times}10^9$, $4.93{\times}10^9$ and $7.46{\times}10^9$ $CFU/m{\ell}$, respectively. The kimchi fermented in the TOV with the most dominant bacterial species were the following 5 strains: Bacillus subtilis, B. licheniformis, B. safensis, Lactobacillus brevis and B. pumilus. The use of different types of containers therefore influenced the number of L. brevis and the four Bacillus species. in kimchi, and may influence the characteristics of the fermented kimchi products. The TOV offered the greatest L. brevis numbers and suggested that it could be the best suited for preparing traditional kimchi fermentation.

Purification and Characterisation of a Burkholderia pseudomallei Protease Expressed in Recombinant E. coli

  • Ling, Jessmi M.L.;Nathan, Sheila;Hin, Lee Kok;Mohamed, Rahmah
    • BMB Reports
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    • v.34 no.6
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    • pp.509-516
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    • 2001
  • A genomic DNA fragment that contains the gene, which codes for a novel extracellular serine protease in Burkholderia pseudomallei, was cloned by using pQE40 as a vector. It was maintained in Escherichia coli JM109. The expression of the gene(s) resulted in the production of a 52 kDa protease. The recombinant protease was purified from the culture filtrate via ammonium sulfate fractionation, gel filtration, and anion-exchange chromatography. The purified protease had an optimum pH and temperature of pH 8.9 and $38^{\circ}C$, respectively. The protease activity was inhibited by EGTA, EDTA, and PMSF, but not 1,10-phenanthroline. The first 11 amino acid residues from the N-terminus of the purified protease were identified as LAPNDPYYYGY. PNDPYY was found to show homology to the Bacillus cereus microbial serine protease and B. subtilis PD498 serine protease. These results indicate that the protease that was purified in this study is an extracellular calcium-dependent serine protease. The purified protease was able to digest the human serum 19A, IgG, albumin, and transferrin, as well as bovine muscle actin and myosin. Furthermore, it was able to promote or cause dermonecrosis in experimental rabbits. These results propose the possible role of a novel B. pseudomallei extracellular calcium-dependent serine protease in the virulence of the pathogen.

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NaCl-dependent Amylase Gene From Badillus circulans F-2 Its Nucleotide Sequence (Bacillus circulans F-2의 NaCl 의존성 amylase 유전자의 DNA 염기배열 결정)

  • 김철호;권석태;타니구치하지메;마루야마요시하루
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.309-316
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    • 1990
  • The sequence of a 1795 bp restriction fragment containing the B. circulans F-2 gene for NaC1- dependent $\alpha$-amylase (CI-amylase) is reported. The probable coding region of the gene is 1005 base pairs (335 amino acida) long. The NaC1-dependent $\alpha$-amylase (el-amy) sequence shows an open reading frame (ORF) with the translated molecular weight of about 38, 006, which correspond to a molecular weight of about 35, 000 (Mi). The gene is preceded by the sequence resembling promoter for the vegetative B, subtitis RNA polymerases. These are followed by the sequences resembling a B. subtilis ribosome binding site 5 nucleotides before the first codon of the gene. Homologous regions with other amylases were found. The N-terminal sequences of the mature proteins expressed in E. eoli were identical to the N-terminal sequences which are anaIysed.

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Antimutagenic effects of browning products reacted with polyphenol oxidase extracted from peach (복숭아 효소 갈변반응 생성물의 돌연변이원성 억제효과)

  • Ham, Seung-Shi;Choi, Kyeong-Kun
    • Applied Biological Chemistry
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    • v.35 no.2
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    • pp.82-86
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    • 1992
  • This research was carried out to investigate antimutagent effect of peach enzymatic browning reaction products(PEBRP) obtained by reacting each of polyphenol compounds with oxidase extracted from Korea-cultivated peach. In methods, rec-assay with B. subtilis strains $H17(rec^+)\;and\;M45(rec^-)$, and Ames test with S. typhimurium TA98 and TA100 were used. The spore rec-assay of PEBRP, pyrogallol, hydroxyhydroquinone, homocatechol and caffeic acid were not showed mutagenicity. In the effects of various metal ions$(Al^{3+},\;Cu^{2+},\;Fe^{2+},\;Mn^{2+},\;Ni^{2+},\;Pb^{2+},\;Zn^{2+})$ on the rec-assay, all PEBRP except caffeic acid was increased inhibition zone(5 mm) only with $Zn^{2+}$. In paticular, the Py-PEBRP was decreased the difference of inhibition zone of growth on MMC(mitomycin C). In results of Ames test, all PEBRP were not showed mutagenicity on S. typhimurium TA98 and TA100; however, Ca-PEBRP and Hca-PEBRP were suppressed mutagenic effects on Trp-P-1 and B(a)P in the presence of S-9Mix.

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Carbon Storage Regulator A (csrA) Gene Regulates Motility and Growth of Bacillus licheniformis in the Presence of Hydrocarbons

  • Angel, Laura Iztacihuatl Serrano;Segura, Daniel;Jimenez, Jeiry Toribio;Barrera, Miguel Angel Rodriguez;Pineda, Carlos Ortuno;Ramirez, Yanet Romero
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.185-192
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    • 2020
  • The global carbon storage regulator (Csr) system is conserved in bacteria and functions as a regulator in the exponential and stationary phases of growth in batch culture. The Csr system plays a role in the central carbon metabolism, virulence, motility, resistance to oxidative stress, and biofilm formation. Although the Csr was extensively studied in Gram negative bacteria, it has been reported only in the control of motility in Bacillus subtilis among Gram positive bacteria. The goal of this study was to explore the role of the csrA gene of Bacillus licheniformis M2-7 on motility and the bacterial ability to use hydrocarbons as carbon source. We deleted the csrA gene of B. licheniformis M2-7 using the plasmid pCsr-L, harboring the spectinomycin cassette obtained from the plasmid pHP45-omega2. Mutants were grown on culture medium supplemented with 2% glucose or 0.1% gasoline and motility was assessed by electron microscopy. We observed that CsrA negatively regulates motility by controlling the expression of the hag gene and the synthesis of flagellin. Notably, we showed the ability of B. licheniformis to use gasoline as a unique carbon source. Our results demonstrated that CsrA is an indispensable regulator for the growth of B. licheniformis M2-7 on gasoline.

Biological Control of Sclerotinia sclerotiorum in Lettuce Using Antagonistic Bacteria (길항세균을 이용한 상추 균핵병의 생물학적 방제)

  • Chon, Bong-Goan;Park, Suji;Kim, Jin-Won
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.12-20
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    • 2013
  • To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

Solubility, Antioxidative and Antimicrobial Activity of Chitosan-Ascorbate (키토산-아스코베이트의 용해성, 항산화성 및 항균성)

  • Lee, Seung-Bae;Lee, Ye-Kyung;Kim, Soon-Dong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.8
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    • pp.973-978
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    • 2006
  • This study was conducted to investigate the solubility, antioxidative and antimicrobial activity of the freeze dried chitosan-ascorbate (CAs) and chitosan-acetate (CAc). In the results of solubility, CAs was soluble over 0.5% in distilled water, vinegar, green tea, soju (distilled liquor), beer and red wine, while it was not soluble in soy sauce, soy milk, milk, orange juice, coffee, sesame oil, soy milk and soybean oil. The solubility of CAc in the liquid foods was similar to those of CAs, but it was soluble less than 0.1% in beer, and formed curd in red wine. Electron donating activity, antioxidative activity and SOD activity of CAs were 48.2, 90.6 and 67.5%, respectively, while the activities of the CAc were 0, 40.0 and 10.0%, respectively. The minimal inhibitory concentrations of CAs and CAc were $200\;{\mu}g/disc$ against Bacillus circulans, Bacillus brevis, Bacillus licheniformis, Bacillus arabitane and Bacillus sterothermophillus, $400\;{\mu}g/disc$ against Escherichia coli O157, Listeria monocytogenous, Bacillus cereus and Bacillus subtilis. There was no significant difference in Hunter's L* value between CAs and CAc $(81.95{\sim}82.97)$, but Hunter's a* and b* values of the CAs was higher than those of CAc. While sour and bitter tastes of CAs were lower than those of CAc, there was no significant difference in astringent taste. From these results, it suggested that CAs has more extensive utility in liquid foods with antimicrobial and antioxidant activity as well as sensory quality compared to CAc.

Antioxidative and Antimicrobial Activities of Aruncus dioicus var. kamtschaticus Hara Extracts (눈개승마(Aruncus dioicus var. kamtschaticus Hara) 용매 추출물의 항산화 및 항균활성)

  • Kim, Mi-Seon;Kim, Kyoung-Hee;Jo, Ji-Eun;Choi, Jong-Jin;Kim, Young-Jin;Kim, Jong-Hwan;Jang, Soon-Ae;Yook, Hong-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.47-55
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    • 2011
  • The solvent extracts of Aruncus dioicus var. kamtschaticus Hara, which were extracted by using several solvents with different polarities, were performed to investigate the antioxidant activities, whitening effect and antimicrobial activity. The content of total polyphenol of fractions from Aruncus dioicus var. kamtschaticus Hara extract showed the highest value ($335.88{\pm}2.26$ mg/g GAE) on ethyl acetate fraction. The ethyl acetate and n-butanol fractions were 0.06 mg/mL and 0.25 mg/mL as $IC_{50}$ values on DPPH radical scavenging, and $99.16{\pm}0.09%$ and $89.29{\pm}0.64%$ on ABTS radical scavenging activity, respectively. Also, reducing power and FRAP value were significantly higher on ethyl acetate fraction. The SOD like activity showed $80.76{\pm}0.61%$ on ethyl acetate and $72.34{\pm}0.79%$ on n-butanol. Tyrosinase inhibition activities (at 5 mg/mL) were $59.08{\pm}0.98%$ on ethyl acetate fraction. The chloroform fraction showed the strongest antimicrobial activities against B. cereus (14 mm), B. subtilis (12.5 mm), S. aureus (10.8 mm), E. coli (20.7 mm) at 0.1 mg/disc and the inhibition zone diameter of ethyl acetate fraction was 17.2 mm against E. coli at 0.5 mg/disc. The minimum inhibitory concentrations (MIC) of chloroform fraction against B. cereus and E. coli were 50 and $25{\mu}g$/mL, respectively. From these results, it is suggested that ethyl acetate and n-butanol fractions of Aruncus dioicus var. kamtschaticus Hara could be used as functional material for food additive ingredient and chloroform fraction could be suitable for the development of a food preservative.

Bacterial Community Profiling during the Manufacturing Process of Traditional Soybean Paste by Pyrosequencing Method (Pyrosequencing을 이용한 전통된장 제조과정 중 세균군집구조의 분석)

  • Kim, Yong-Sang;Jeong, Do-Yeon;Hwang, Young-Tae;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.275-280
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    • 2011
  • In order to evaluate the diversity and change of bacterial population during the manufacturing process of traditional soybean paste (doenjang), bacterial communities were analyzed using 16S rRNA gene-based pyrosequencing. In rice straw, the most important inoculum source for fermentation, the bacterial sequences with a relative abundance greater than 1% were assigned to four phyla, Proteobacteria (71%), Actinobacteria (20.6%), Bacteroidetes (4.2%), and Firmicutes (1.3%). Unlike bacterial community composition of rice straw, a different pattern of bacterial population in meju was observed with predominantly high abundance (99.1%) of Firmicutes. Phylum composition in young doenjang was almost same as that of meju. Major genera in young doenjang were Bacillus (81.3%), Clostridium (6.9%) and Enterococcus (6.3%) and the predominant species among bacterial population was B. amyloliquefaciens (63.6%). Abundance of the phylum Firmicutes in mature doenjang was 99.98%, which was even higher value than those in meju and young doenjang. Predominant species in mature doenjang were B. amyloliquefaciens (67.3%), B. atrophaeus (12.7%), B. methylotrophicus (6.5%), B. mojavensis (3.2%), and B. subtilis. (2.5%), which were also identified as major species of the microbial flora in meju. These results suggested that rice straw was a primary source for supplement of Bacillus species in manufacturing the traditional doenjang and that some species of Bacillus strains were mainly involved in the fermentation process of traditional doenjang.

Evaluation of Environment-friendly Control Agents for the Management of Powdery Mildew Infection during Seedling Stage of Three Cucurbitaceae Vegetables (친환경 육묘시 세 가지 박과채소의 흰가루병에 대한 친환경 제제의 방제효과)

  • Yeo, Kyung-Hwan;Jang, Yoon Ah;Kim, Su;Um, Young Chul;Lee, Sang Gyu;Rhee, Han Cheol
    • Journal of Bio-Environment Control
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    • v.22 no.4
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    • pp.413-420
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    • 2013
  • The purpose of this study was to evaluate the control efficacy of major environment-friendly control agents against powdery mildew, as affected by the application condition such as disease-developing stage and microclimate, as compared with the efficacy of pesticides in plug seedling of three Cucurbitaceae vegetables, including cucumber, melon, and oriental melon. Single or combined application of major six environment-friendly control agents was used in the experiment: two biofungicide (Ampelomyces quisqualis 94013 and Bacillus subtilis Y1336), two plant extracts (neem oil and extracts of Rheum undulatum), and two mineral materials (wettable sulfur powder and lime bordeaux mixture). These control agents were treated to the plug seedlings for preventing powdery mildew and curative applications for managing powdery mildew. In all treatments, the disease incidence declined as daily average temperatures increased to $30^{\circ}C$ for consecutive 6~8 days with maximum temperature over $40^{\circ}C$. In preventative application, the control efficacy against powdery mildew was the highest in the treatment of wettable sulfur powder, and lowest in the B. subtilis Y1336, with values of 20~40%. In cucumber seedlings, the preventive single application of neem oil or wettable sulfur powder was more effective than curative application of fungicides, while the control efficacy of these agents was similar to those of fungicides in melon seedlings. The single application of R. undulatum extracts was also effective in preventing the disease for both cucumber and melon seedlings, showing a higher control efficacy than those of biofungicides during seedling stage. The treatment of water spray was not effective and showed a higher disease incidence than the untreated control plot in the oriental melon and melon seedlings. The curative application with environment friendly control agents, after powdery mildew was first detected, could not successfully controlled the disease at the middle stage (5~10% of disease incidence) of disease development. The curative combined application of [R. undulatum extracts (1st application) + wettable sulfur powder (2nd) + neem oil (3rd)] showed the highest control efficacy among the other treatments, with control value over 80% at the early stage (less than 1% of disease incidence) of disease development.