• Title/Summary/Keyword: Microbial reaction

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Purification and Properties of Protease from Thermophilic Actinomyces (고온성 방선균이 생산하는 단백질 분해효소의 정제와 특성)

  • 김중배
    • The Korean Journal of Food And Nutrition
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    • v.13 no.2
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    • pp.176-180
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    • 2000
  • Microbial protease has been interesting due to the biological roles in the producing microorganism. A thermophilic Actinomyces produing protease was isolated from soil. The optimal medium composition and culture conditions for maximum protease production was as follows 0.5% soluble starch, 0.5% yeast extract. 0.1% K2HPO4, 0.05% CaCl2, initial pH 8.0 at 50$^{\circ}C$ for 48hours. The protease was purified by the procedure of ammonium sulfate precipitation, anion exchange chromatography(LC), DEAE high performance liquid chromatography and GPC HPLC. The purification fold of the purified enzyme was increased about 22.6. The optimal pH and temperature for reaction of the purified enzyme were 7.5 and 60$^{\circ}C$. The purified enzyme was stable for the pH range from 6.0 to 8.5, but was unstable when treated at 80$^{\circ}C$ for 10 minutes. The activity of the enzyme was inhibited by Ag+ and Cu2+.

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유기용매 내에서 중쇄지방질의 합성

  • Gwon, Dae-Yeong
    • Bulletin of Food Technology
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    • v.7 no.2
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    • pp.64-73
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    • 1994
  • Using 20 lipases from various microbial origins medium chain glycerides, namely, mono-, di-, and tri-carproyl glycerols from glycerol and acid were synthesized in isooctane. Enzyme reaction was performed at 0.35 M of capric acid, 0.025M of glycerol and the same mass of silica gel to remove water in 5ml of isooctane with 30mg of lyophilized lipase. Among 20 lipases, eleven lipases showed good synthetic activities, especially lipase from Pseudomonas aeruginosa (Lipase PS), Rhizomucor miehei origined lipase and Chromobacterium viscosum lipase (Lipase CV) showed good activities for production of tricaproylglycerol, while Lipase OF-360 (origined from Candida rugosa) and Lipase D (Rhizopus delemar) were good for production of dicaprolyglycerol. The lipases, especially Lipase PS, have high thermal stability at $ 60^{circ}C$, and optimum pH of lyophilization for dehydrating the lipase was pH 6.5.

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The Effect of Barbaloin on LPS-stimulated Inflammatory Reaction in Mice Peritoneal Macrophages

  • Jeon, Yong-Deok;Lee, Jong-Hyun
    • Korean Journal of Plant Resources
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    • v.30 no.3
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    • pp.280-286
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    • 2017
  • Barbaloin is a major component of Aloe vera, which has been used for a laxative. Also, barbaloin is C-glucoside of aloe emodin anthrone which is founded in Aloe vera. Barbaloin has varieties of pharmacological activity such as inhibitory effects on inflammation, histamine release, cancer and microbial infection. But the effect of barbaloin on lipopolysaccharide (LPS)-stimulated macrophages has not been understood. In this study, we evaluated the effects of barbaloin against LPS-stimulated production of nitric oxide (NO), inflammatory cytokines and MAPKs activation in macrophage. We treated barbaloin (0.1, 1, 10, $100{\mu}M$) in LPS-stimulated mice peritoneal macrophage. Our results showed that barbaloin significantly inhibited production of NO and cytokines of tumor necrosis factor $(TNF)-{\alpha}$, interleukin (IL)-6, interleukin $(IL)-1{\beta}$ in LPS-stimulated peritoneal macrophage. Moreover, barbaloin inhibited the phosphorylation of ERK and JNK in a dose dependent manner. These results indicated that barbaloin could be useful for inflammatory diseases.

Isolation of Urease Positive Vibrio parahaemolyticus and Urease Production (Urease를 생산하는 Vibrio parahaemolyticus의 분리 및 urease 생산)

  • 김종숙;김영희
    • Journal of Life Science
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    • v.10 no.1
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    • pp.94-100
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    • 2000
  • Urease is an important microbial enzyme and its production is a marker to predict potential pathogenicity. An unusual halophilic bacterium producing urease was isolated from sea product and identified as Vibrio parahaemolyticus KH410. Its biochemical properties were indole negative, gelatin positive, sodium citrate positive and Kanagawa positive whereas other characteristics were identical as the standard strain except it showed a positive reaction on Christensen's urea agar. V. parahaemolyticus urease production was directly related to urea concentration. The production of urease was noticeable by the addition of 0.2% urea, 0.5% glucose, 2% NaCl in LB broth, and the initial pH of 5.5. The maximum production reached after 6 hr of incubation at 37$^{\circ}C$. However, NiCl2, metal ions, phosphorus did not affect production of urease.

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CHARACTERISTICS OF BIOHYDROGEN PRODUCTION AND MICROBIAL COMMUNITY AS A FUNCTION OF SUBSTRATE CONCENTRATION

  • Youn, Jong-Ho;Shin, Hang-Sik
    • Environmental Engineering Research
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    • v.10 no.1
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    • pp.7-14
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    • 2005
  • The feasibility of hydrogen production with a raw seed sludge through direct acclimation of feedstock was investigated at acidogenic stage, and methane was harvested at followed methanogenic stage in an anaerobic two-stage process. Hydrogen content was higher than 57% at all tested organic loading rates (OLRs) and the yield of hydrogen ranged from 1.5 to 2.4 mol H2/mol hexose consumed and peaked at 6 gVSl-1day-1. Normal butyrate and acetate were main volatile fatty acids (VFAs), whereas the concentration of propionate was insignificant. The hydrogen-producing bacteria, Clostridium thermosaccharolyticum, was detected with strong intensity at all tested organic loading rates (OLRs) by denaturing gradient gel electrophoresis (DGGE) of the polymerase chain reaction (PCR) analysis. From COD balance in the process, the fraction of the feed-COD converted to the hydrogen-COD at acidogenic stage ranged from 7.9% to 9.3% and peaked at 6 gVSl-1day-1, whereas the fraction of feed-COD converted to the methane-COD at methanogenic stage ranged from 66.2% to 72.3% and peaked at 3 gVSl-1day-1.

A Study on Optimum Moisture Content and C/N ratio of Sewage Sludge Treatment Using Composting (퇴비화를 이용한 하수슬러지 처리에 있어서 적정 수분함량과 C/N비에 관한 연구)

  • 손현석;양원호;정문식
    • Journal of Environmental Health Sciences
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    • v.23 no.2
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    • pp.44-56
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    • 1997
  • When sewage sludge is treated by cornposting, higher moisture content and lower C/N ratio on sewage sludge is problems. This paper project to alesolve two problems. The almost trends in run 3 of MC 70% are similar to these in run 1 and 4 of MC 65%. A retention time of the highest temperature (>50$\circ$C) and increase rate of temperature in run 3 are an affinity to these in run 4. Particularly, decrease rate of temperature in run 3 is slower than others and this data shows the more activity of thermal microbial in run 3 than that in others. C/N ratio trends in run 3 shows slow reaction in initial stage but, after 9 days, is similar to that in run 1 and 4. Carbon trends in each run are a similarity to C/N ratio trends. Temperature, MC, carbon and C/N ratio trends in run 5, whose C/N ratio is 15, show less microbial activity than that in run 6, whose C/N ratio is 20. But temperature increase of the beginning stage and pH of the final stage in run 5 are greater than that in run 6. Final MC and carbon content in run 5 and 6 have a similar values. That is, final MC in run 5 and 6 is 49.39% and 48.97% and final carbon content in each run is 25.15% and 22.20%. Expecially, a temperature increase and C/N ratio decrease rate of the beginning stage in run 5 are greater than these in run 6. This shows the shorter lag time in run 5 than lag time in run 6.

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Development of a Species-specific PCR Assay for Three Xanthomonas Species, Causing Bulb and Flower Diseases, Based on Their Genome Sequences

  • Back, Chang-Gi;Lee, Seung-Yeol;Lee, Boo-Ja;Yea, Mi-Chi;Kim, Sang-Mok;Kang, In-Kyu;Cha, Jae-Soon;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.212-218
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    • 2015
  • In this study, we developed a species-specific PCR assay for rapid and accurate detection of three Xanthomonas species, X. axonopodis pv. poinsettiicola (XAP), X. hyacinthi (XH) and X. campestris pv. zantedeschiae (XCZ), based on their draft genome sequences. XAP, XH and XCZ genomes consist of single chromosomes that contain 5,221, 4,395 and 7,986 protein coding genes, respectively. Species-specific primers were designed from variable regions of the draft genome sequence data and assessed by a PCR-based detection method. These primers were also tested for specificity against 17 allied Xanthomonas species as well as against the host DNA and the microbial community of the host surface. Three primer sets were found to be very specific and no amplification product was obtained with the host DNA and the microbial community of the host surface. In addition, a detection limit of $1pg/{\mu}l$ per PCR reaction was detected when these primer sets were used to amplify corresponding bacterial DNAs. Therefore, these primer sets and the developed species-specific PCR assay represent a valuable, sensitive, and rapid diagnostic tool that can be used to detect three specific pathogens at early stages of infection and may help control diseases.

Relationship of Oral Bacterial Load Over One Year of Smoking Cessation

  • Kim, Sunghyun;Seo, Min-Seock;Hwang, Soo-Jeong
    • Journal of dental hygiene science
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    • v.19 no.4
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    • pp.213-219
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    • 2019
  • Background: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking. Methods: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient. Results: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF. Conclusion: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.

PCR Method Based on the ogdH Gene for the Detection of Salmonella spp. from Chicken Meat Samples

  • Jin, Un-Ho;Cho, Sung-Hak;Kim, Min-Gon;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Kim, Kwang-Yup;Chung, Duck Hwa;Lee, Young-Choon;Kim, Cheorl-Ho
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.216-222
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    • 2004
  • In a previous paper, the ogdH gene that encodes 2-oxoglutarat dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-l and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method.

Effect of Low Salt Concentrations on Microbial Changes During Kimchi Fermentation Monitored by PCR-DGGE and Their Sensory Acceptance

  • Ahmadsah, Lenny S. F.;Min, Sung-Gi;Han, Seon-Kyeong;Hong, Yeun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2049-2057
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    • 2015
  • Various salt concentrations (1.0%, 1.3%, 1.6%, 1.9%, and 2.1% labeled as sample A, B, C, D, and E, respectively) were investigated for microbial diversity, identification of Lactic Acid Bacteria (LAB) in salted kimchi cabbage, prepared under laboratory conditions. These samples were stored at 4°C for 5 weeks in proper aluminum-metalized pouch packaging with calcium hydroxide gas absorber. A culture-independent method known as polymerase chain reaction - denaturing gradient gel electrophoresis was carried out to identify LAB distributions among various salt concentration samples that had identified 2 Weissella (W. confusa and W. soli), 1 Lactobacillus (Lb. sakei), and 3 Leuconostoc (Lc. mesenteroides, Lc. lactis, and Lc. gelidum) in the overall kimchi samples. The pH, titratable acidity, viable cell counts, and coliform counts were not affected by salt variations. In order to assess sensory acceptance, the conducted sensory evaluation using a 9-point hedonic scale had revealed that samples with 1.3% salt concentration (lower than the manufacturer's regular salt concentration) was more preferred, indicating that the use of 1.3% salt concentration was acceptable in normal kimchi fermentation for its quality and safety. Despite similarities in pH, titratable acidity, viable cell counts, coliform counts, and LAB distributions among the various salt concentrations of kimchi samples, the sample with 1.3% salt concentration was shown to be the most preferred, indicating that this salt concentration was suitable in kimchi production in order to reduce salt intake through kimchi consumptions.