• Title/Summary/Keyword: Stationary Phase

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A Large Scale Separation & Purification of Cyanobacterial Toxins (남조류 독성물질의 대량분리 및 정제)

  • Yoon, Suk Chang;Park, Keun Young;Pyo, Dong Jin
    • Journal of the Korean Chemical Society
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    • v.42 no.1
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    • pp.51-56
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    • 1998
  • It is very difficult to separate and purify the microcystins, cyanobacterial toxins since it exist in a trace level in natural lakes. In this paper, we developed a new analytical method to separate and purify the microcystin RR and LR from the freeze-dried cyanobacterial cells in natural lakes. We used 7.5 g silica gel as a stationary phase and ethyl acetate: isopropanol: water (30: 45: 25) as a mobile phase and microcystins were eluted using an open column. The eluting solvent was collected in a small bottle at the intervals of 3 mL and the fractions were chromatographed with HPLC to confirm the microcystin RR and LR.

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Characterization of an Amylase-sensitive Bacteriocin DF01 Produced by Lactobacillus brevis DF01 Isolated from Dongchimi, Korean Fermented Vegetable

  • Kang, Tae-Kyu;Kim, Wang-June
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.795-803
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    • 2010
  • A DF01 strain that inhibits tyramine-producing Lactobacillus curvatus KFRI 166 was isolated from Dongchimi, a traditional Korean fermented vegetable, and identified as Lactobacillus brevis by biochemical analysis and reverse transcriptase sequencing of 16S rRNA. The antimicrobial compound produced by L. brevis DF01 was secreted at a maximum level of 640 AU/mL in late exponential phase in MRS broth, and its activity remained constant during stationary phase. The activity of bacteriocin DF01 was totally inactivated by $\alpha$-chymotrypsin, pronase E, proteinase K, trypsin, and $\alpha$-amylase, but not by catalase, which indicates the compound was glycoprotein in nature. The activity was not affected by pH changes ranging from 2 to 12 or heat treatment (60, 80, and $100^{\circ}C$ for 30 min), but was reduced after autoclaving. Bacteriocin DF01 had bacteriolytic activity and a molecular weight of approximately 8.2 kDa, as shown by tricine-SDS-PAGE analysis. Therefore, bacteriocin DF01 can be used in the manufacture of fermented meat products due to its inhibition of tyramine-producing L. curvatus and non-inhibition of L. sake, which is used as a starter culture for meat fermentation.

Characterization of Immobilized Denitrifying Bacteria Isolated from Municipal Sewage

  • Kim, Joong-Kyun;Kim, Sung-Koo;Kim, Sang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.756-762
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    • 2001
  • As a component for a recirculating aquaculture system, a new strain of denitrifying bacterium was isolated from municipal sewage. The isolate was motile by means of one polar flagellum, catalase-positive, and a Gram-negative rod-shaped cell measuring $0.5-0.6{\mu}m$ in width and $1.3-1.9{\mu}m$ in length. The isolate was identified as Pseudomonas fluorescens and produced dinitrogen gas via the reduction of nitrate. The optimal growth conditions (pH, temperature, carbon source, and C/N ratio) of the isolate were found to be 6.8, $30^{\circ}C$, malate, and 3, respectively. Under optimal growth conditions of P. fluorescens, dinitrogen gas was first detected in the exponential growth phase, then a small amount of nitrite was developed and converted to dinitrogen gas in the stationary phase. Pseudomonas fluorescens cells were immobilized in modified polyvinyl alcohol (PVA) gel beads, and the maximum denitrification rate was measured as $36.6 {\mu}lN_2h^-1$ per bead with an optimum cell loading of $20mg {\mu}l^-1$ and $2\%$ sodium alginate added to the PVA gel. The operating stability of the modified PVA gel beads remained unchanged for up to 43 repeated batches.

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Characterization of Antimicrobial Substance Producing Lactococcus sp. HM58 Isolated from Gastrointestinal Track of Flounder

  • Jeong Hyun-Mi;Yum Do-Young;Lee Jung-Ki;Choi Mi-Young;Kim Jin-Man
    • Fisheries and Aquatic Sciences
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    • v.6 no.1
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    • pp.27-33
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    • 2003
  • A lactic acid bacterium showing antimicrobial activity against fish pathogen was isolated from gastrointestinal tract of flounder for the purpose of use as an aquaculture probiotics. From the analysis of morphological and physiological characteristics, the isolated strain was named as Lactococcus sp. HM58. Antimicrobial substance (AMS) from Lactococcus sp. HM58 showed strong growth inhibitory activity against Streptococcus sp., which is a fish pathogenic bacterium. AMS was presumed a proteinaceous compound with stability in heat and wide pH range from 2 to 10. It was started to produce in exponential growth phase and was not produced any more in stationary phase. It showed comparatively broad antimicrobial spectrum against most of gram positive bacteria used for this study. About $84\%$ of Lactococcus sp. HM58 was able to survive in the artificial gastric juice though it was low to the extent in the artificial bile juice. In the sensitivity test for various antibiotics, this strain was highly sensitive for doxycycline, erythromycin, amoxicillin clavu1anic acid and ampicillin.

Method development and validation for dieckol in the standardization of phlorotannin preparations

  • Kim, Jiyoung;Um, Minyoung;Yang, Hyejin;Kim, Inho;Lee, Changho;Kim, Yuntai;Yoon, Minseok;Kim, Youngkyoung;Kim, Jimi;Cho, Suengmok
    • Fisheries and Aquatic Sciences
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    • v.19 no.1
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    • pp.3.1-3.6
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    • 2016
  • Phlorotannins are reported to have diverse biological properties. However, no analytical methods for the standardization of phlorotannin preparations have been reported. Herein, we developed and validated an analytical method for the determination of dieckol in phlorotannin extracts (PRT) using high-performance liquid chromatography (HPLC). The optimum HPLC conditions consisted of a Supelco Discovery C18 column stationary phase, a mobile phase (A: 15 % HPLC grade methanol in deionized water, B: methanol), UV detection at 230 nm, and a flow rate of 0.7 mL/min. The optimized chromatographic conditions were validated and exhibited good specificity and linearity ($R^2$ > 0.9994-1.0000). The recoveries were in the range of 100.9-102.3 %. The method had good intermediate (%RSD 1.2) and intra-day (%RSD 0.4-1.7) assay precisions. This HPLC method had good accuracy and quality in the determination of dieckol in PRT.

Isolation and Characterization of a Phenol-Degrading Strain Acinetobacter sp.GEM2 (Phenol을 분해하는 Acinetobacter sp. GEM2의 분리 및 특성)

  • Lee, Chang-Ho;Oh, Hee-Mock;Kwon, Tae-Jong;Kwon, Gi-Seok;Lee, Sung-Gie;Suh, Hyun-Hyo;Yoon, Byung-Dae
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.692-699
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    • 1994
  • A bacterial strain which formed a distinct colony on agar plate containing phenol as a vapor phase and grew well in a liquid minimal medium was isolated and identified as Acinetobac- ter sp. GEM2. The optimal temperature and initial pH for the growth of Acinetobacter sp. GEM2 were 30$\circ$C and 7.0, respectively. Cell growth was inhibited by phenol at the concentration over 1500 ppm. Cell growth dramatically increased from 10 hours after cultivation and almost showed a stationary phase within 24 hours at which 95% of phenol was concomitantly degraded. Acinetobac- ter sp. GEM2 was capable of growing on aromatic compounds, such as benzoic acid, phenol, m- cresol, o-cresol, P-cresol, catechol, gentisic acid, and toluene, but did not grow on benzene, salicylic acid, p-toluic acid, and p-xylene. By the analysis of catechol dioxygenase, it seemed that catechol was degraded through both meta- and ortho-cleavage pathway. The growth-limiting log P value of Acinetobacter sp. GEM2 on organic solvents was 2.0.

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The Influence of Two Phase Flow on Fretting Wear between Steam Generator Tube and Supporting Bar (이상 유동 환경이 증기 발생기 세관과 지지대의 프레팅 마모에 미치는 영향에 대한 연구)

  • Lee, Young-Ze;Park, Jung-Min;Jeong, Sung-Hoon;Kim, Jin-Seon;Park, Se-Min
    • Tribology and Lubricants
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    • v.24 no.6
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    • pp.362-367
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    • 2008
  • Tubes in nuclear steam generators are held up by supports because the tubes are long and slender. Fluid flows of high-pressure and high-temperature in the tubes cause oscillating motions between tubes and supports. This is called as FIV (flow induced vibration), which causes fretting wear in contact parts of tube-support. The fretting wear of tube-support can threaten the safety of nuclear power plant. The tube and support materials were Inconel 690 and STS 409. The wear tests were conducted in various environments, which are in water without flow, in flowing water and in flowing water with air. The results showed that the flow of water influenced on the wear-life of tube. The wear-life of tube decreased in water flow as compared with wear-life in stationary water.

Effects of Cultivation Conditions on the Growth and Polyamine Composition in Methylobacterium extorquens AM1 Growing on Methanol (Methylobacterium extorquens AM1의 메탄올을 이용한 성장과 세포내 폴리아민 구성에 미치는 배양조건의 영향)

  • 엄치용;박기정;강빈구;김영민
    • Korean Journal of Microbiology
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    • v.29 no.6
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    • pp.387-391
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    • 1991
  • Methylobacterium extorquens AM1 growing on methanol as a sole source of carbon and energy was found to grow most rapidly (t$t_{d}$ =6h) at 30.deg.C in a mineral medium (pH 7.0) containing 0.5% (v/v) methanol which was agitated at 200 rpm (optimal cultivation condition). Cells grown under the optimal cultivation condition contained more spermidine, but less putrescine, than the cells grown on 2.5%(v/v) ( $t_{d}$ =8h ) or at 20.deg.C ( $t_{d}$ =8h ). Cells cultivated under the optimal condition was found to contain more spermidine than the cells grown at pH 6.0 (( $t_{d}$ =7h ) and pH 8.0 ($t_{d}$ =7.3h). the cells growing at the stationary phase under the optimal condition accumulated more spermine or putrescine than the cells growing at the same phase on 2.5%(v/v) methanol or at pH 6.0 and pH 8.0, respectively. M. extorquens AM1 grown in a medium agitated at 100 rpm ( $t_{d}$ =8.8h ) contained less spermidine and spermine than the cells grown under the optimal cultivation condition.

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Efficient Expression of a Carbon Starvation Promoter Activity Under Nutrient-Limited Chemostat Culture

  • KIM DAE-SUN;PARK YONG-IL;LEE HYANG BURM;KIM YOUNGJUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.678-682
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    • 2005
  • The promoter region of a carbon starvation gene isolated from Pseudomonas putida was cloned and analyzed for its potential use for in situ bioremediation and bioprocessing. We constructed a recombinant plasmid pMKD101 by cloning the 0.65 kb promoter region of the gene into the promoter proving vector, pMK301, which contains the lacZ for ${\beta}$-galactosidase activity as a reporter gene. pMKD101 was transformed into the wild-type P. putida MK1, resulting in P. putida RPD101, and analyzed for ${\beta}$-galactosidase activity under different culture conditions. When RPD101 was grown on the minimal medium plus $0.1\%$ glucose as a sole carbon source in batch cultures, ${\beta}$-galactosidase activity was found to be 3.2-fold higher during the stationary phase than during the exponential phase. In chemostat cultures, ${\beta}$-galactosidase activity was found to be 3.1-fold higher at the minimal growth rate (dilution rate=$0.05\;h^{-1}$) than at the maximal growth rate (dilution rate=$0.173;h^{-1}$). The results suggest that a carbon starvation promoter can be utilized to maximize the expression of a desired gene under nutrient limitation.

Characterization of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju

  • Lim, Sung-Mee;Park, Mi-Yeon;Chang, Dong-Suck
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.49-57
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    • 2005
  • Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.