• Title/Summary/Keyword: stationary phase

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Culture of Tricholoma matsutake Mycelium using Solid Matrix (고체 매질을 이용한 송이(松栮)(Tricholoma matsutake)균 배양)

  • Lee, Wi Young;Ahn, Jin Kwon;Ka, Kang Hyeon;Park, Hyun
    • Journal of Korean Society of Forest Science
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    • v.95 no.3
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    • pp.358-364
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    • 2006
  • This study was conducted to develop optimal solid culture medium for Tricholoma matsutake. As the solid matrix, granitic soil, perlite, vermiculate, pine sawdust and peat moss were compared regarding their effected on mycelial growth. Ergosterol content which is a fungal wall component was used as the growth index of the mycelia. Among the various solid matrixes, the granitic soil, perlite and mixture of the two supported the growth most. Barely flour appeared to be very effective on the stimulating of the mycelial growth when added to the solid matrix. An mixture of the matrix contained an even (1:1:1:1, v/v/v/v) mixture of granitic soil, perlite, vermiculate and pine sawdust. T. matsutake started growth 2 weeks after inoculation and reached stationary growth phase after 8th weeks in the solid matrix mixture. The mycelial density in the solid matrix was 7 times higher than that in fairy-ring soil. In addition, 30~70% water content and 10% humus soil in the solid matrix also supported good growth suggesting that T. matsutake needs humus soil for a nutrient sources. The solid matrix developed in the present study could be used to study physiological characteristics of T. matsutake as well.

Purification and Characterization of a Fibrinolytic Enzyme Produced by Bacillus amyloliquefaciens HC188 (Bacillus amyloliquefaciens HC188이 생산하는 혈전분해 효소의 정제 및 특성)

  • Shin, So Hee;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.33-43
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    • 2013
  • A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Physiological and Ecological Characteristis of Hemolytic Vibrios and Development of Sanitary Countermeasure of Raw Fisheries Foods. 2. Physiological and Psychrotrophic Characteristics of Vibrio mimicus SM-9 Isolated from sea Water (용혈독소를 생산하는 기수성 비브리오균의 생리.생태적 특성과 수산식품의 위생대책 2. 해수에서 분리된 vibrio mimicus SM-9의 생리적 특성 및 저온내성)

  • 장동석;김신명;박욱연;박미연;김영만
    • Journal of Food Hygiene and Safety
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    • v.12 no.1
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    • pp.9-14
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    • 1997
  • Vibrio mimicus is a closely related species with V. cholerae, and has been reported to be associated with gastrointestinal infections. Although extraintestinal infections of these vibrios have also been reported in Japan and Southeast Asia. But little research papers on V. mimicus was reported in Korea. Therefore, we tried to isolate V. mimicus from the environmental sea water from April to July in Pusan, Korea. Among the isolated strains, we selected the strongest hemolytic strain and then named V. mimicus SM-9. In this paper, we checked the antibiotic susceptibility and psychrotrophic characteristics of the isolated strain. Hemolytic activity of the hemolysin produced by the isolated strain was also measured. V. mimicus was not detected from the sea water samples in April and May, but its detection rate was relatively high in June and July in Pusan, Korea. The bacteriological characteristics of V. mimicus SM-9 were Gram-negative rods, motile, oxidase positive, Voges-Proskauer negative and sucrose negative. In 23 kinds of antibiotics susceptibility test, V. mimicus SM-9 showed susceptibility to the most of antibiotics submitted while it was resistive against lincomycin, oxacillin, rifampin and vancomycin. Hemolytic activity of the hemolysin produced by V. mimicus SM-9 was highest in stationary phase of the growth curve in BHI broth at 37$^{\circ}C$ and its activity was reached 18 HU per $m\ell$ of culture supernatant. For checking the psychrotrophic property of V. mimicus SM-9, the decreasing rate of the strain in phosphate buffer solution and yellowtail flesh homogenate was examined during the storage at 4, 0, -4 and -2$0^{\circ}C$. The decreasing rates of the selected strain stored in phosphate buffer solution were greater than those in fish homogenate. Decreasing rates of V. mimicus SM-9 stored in phosphate buffer solution were not significantly different by the storage temperatures. The viable cell counts of the strain were decreased as 5 log cycles after 120 hours at all the tested temperatures. While decreasing numbers of the strain in fish homogenates were 2*4 log cycles after 120 hours. The decreasing pattern of the strain numbers were very slow after 200 hours at all the stored temperatures.

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Analysis of Molecular Species of Vegetable Oil Triglycerides by Capillary Column GC-MS (Capillary Column GC-MS에 의한 식물유 트리글리세리드 분자종의 분석)

  • Yoon, Hyeung-Sik;Kim, Seon-Bong;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.21 no.3
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    • pp.391-398
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    • 1989
  • Triglyceride molecular species In some vegetable oils were analyzed by capillary column gas chromatography and electron impact ionization mass spectrometry utilizing selected ion monitoring. Triglycerides were separated according to their molecular weights and their degrees of unsaturation on $25m{\times}0.25mm$ fused silica open tubular capillary column coated with a phenylmethylsilicone gum stationary phase and in an analysis time less than 13 min. Triglyceride molecular species were identified by analyzing the fragment ions having the same time on the selected ion monitoring profile . The major triglyceride molecular species in each oils were $C_{18:1}.\;C_{18:2}.\;C_{18:2}(OLL:18.3%),\;C_{18:2}.\;C_{18:2}.\;C_{18:2}(LLL;\;14.3%),\;C_{18:0}.\;C_{18:2}.\;C_{18:2}(SLL;\;14.1%),\;C_{16:0}.\;C_{18:2}.\;C_{18:2}(PLL;\;13.2%),\;C_{16:0}.\;C_{18:2}.\;C_{18:1}(PLO;\;11.6%)$ in corn oil, $C_{18:2}.\;C_{18:2}.\;C_{18:2}(LLL;\;18.0%),\;C_{18:1}.\;C_{18:2}.\;C_{18:2}(OLL;\;18.0%),\;C_{16:0}.\;C_{18:2}.\;C_{18:2}(PLL;\;17.1%)$ in safflower oil, $C_{16:0}.\;C_{18:2}.\;C_{18:2}(PLL;\;23.5%),\;C_{16:0}.\;C_{18:2}.\;C_{18:1}(PLO;\;13.8%),\;C_{18:0}.\;C_{18:1}.\;C_{18:1}(SOO;\;13.5%),\;C_{18:1}.\;C_{18:2}.\;C_{18:2}(OLL;\;10.6%)$ in cottonseed oil.

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Comparison of Biomechanical Characteristics of Rowing Performance between Elite and Non-Elite Scull Rowers: A Pilot Study

  • Kim, Jin-Sun;Cho, Hanyeop;Han, Bo-Ram;Yoon, So-Ya;Park, Seonhyung;Cho, Hyunseung;Lee, Joohyeon;Lee, Hae-Dong
    • Korean Journal of Applied Biomechanics
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    • v.26 no.1
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    • pp.21-30
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    • 2016
  • Objective: This study aimed to examine the characteristics of joint kinematics and synchronicity of rowing motion between elite and non-elite rowers. Methods: Two elite and two non-elite rowers performed rowing strokes (3 trials, 20 strokes in each trial) at three different stroke rates (20, 30, 40 stroke/min) on two stationary rowing ergometers. The rowing motions of the rowers were captured using a 3-dimensional motion analysis system (8-infrared camera VICON system, Oxford, UK). The range of motion (RoM) of the knee, hip, and elbow joints on the sagittal plane, the lead time ($T_{Lead}$) and the drive time $T_{Drive}$) for each joint, and the elapsed time for the knee joint to maintain a fully extended position ($T_{Knee}$) during the stroke were analyzed and compared between elite and non-elite rowers. Synchronicity of the rowing motion within and between groups was examined using coefficients of variation (CV) of the $T_{Drive}$ for each joint. Results: Regardless of the stroke rate, the RoM of all joints were greater for the elite than for non-elite rowers, except for the RoMs of the knee joint at 30 stroke/min and the elbow joint at 40 stroke/min (p < .05). Although the $T_{Lead}$ at all stroke rates were the same between the groups, the $T_{Drive}$ for each joint was shorter for the elite than for the non-elite rowers. During the drive phase, elite rowers kept the fully extended knee joint angle longer than the non-elite rowers (p < .05). The CV values of the TDrive within each group were smaller for the elite compared with non-elite rowers, except for the CV values of the hip at all stroke/min and elbow at 40 stroke/min. Conclusion: The elite, compared with non-elite, rowers seem to be able to perform more powerful and efficient rowing strokes with large RoM and a short $T_{Drive}$ with the same $T_{Lead}$.

Free Radical Scavenging Activity and Ascorbate Content in Various Plant Cell Lines (다양한 식물배양세포주에서 자유라디칼 포착활성과 Ascorbate 함량)

  • AHN, Young-Ock;CHOI, Yong-Hwa;KWON, Seok-Yoon;LEE, Haeng-Soon;KIM, Suk-Won;PARK, Il-Hyun;KWAK, Sang-Soo
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.289-293
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    • 1998
  • We investigated the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in methanol extracts of 64 cultured cell lines, which were derived from various plant species, and the ascorbate content in cell lines, which showed a high radical scavenging activity. Thirteen cell lines revealed the antioxidative activity ($IC_{50}$) by methanol extracts of less than 50 mg in cell fresh wt. Of them, six cell lines showed the same Rf value as ascorbate on the DPPH sprayed silica gel TLC. The ascorbate content in cell lines of Rosa multiflora, Scutellaria baicalensis, and Achyranthes japonica showed 48.5, 30.3, and $16.8\;\mu\textrm{g}$ per g cell fresh wt by HPLC analysis, respectively. In callus cultures of S. baicalensis, the concentration of ascorbate reached a maximun ($39{\pm}3.4\;\mu\textrm{g}/g$ cell fresh wt) on 30 days after subculture, which corresponded to the stationary growth phase, and subsequently decreased by successive culturing.

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Optimization of Indole-3-Acetic production by phosphate solubilization bacteria isolated from waste mushroom bed of Agaricus bisporus

  • Walpola, Buddhi Charana;Noh, Jae-Geun;Kim, Chan Kyem;Kyung, Ki-Cheon;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.53-62
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    • 2013
  • A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$ $ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$ $ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.

Isolation and Characterization of Cryptococcus sp. CS-2 Secreting Polygalacturonase from Soil (토양으로부터 Cryptococcus sp. CS-2의 분리 및 균주가 분비하는 Polygalacturonase의 특성에 관한 연구)

  • 강희경;문명님;임채영;양영기
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.158-163
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    • 1999
  • A ploygalacturonase-produchg yeast was isolated from Cheju soil by selective eivichment media. One strain which has the highesl activity of polygalacturonase was selected. The characle~ishcs of the strain CS-2 were as follows: CS-2 utilized xylose. sucrose, maltose, u.ehalose, cellobiose. melibiose, lactose, raffinose, inosiiol, dulicilol, and dextrose, but did not utilized galactose, nitrate. nit~te, and lysine. Growth of CS-2 was inhibited by cyclohexamide, 1% acetic acid, and high concenaation (over 50%) of glucose. It grew at $30^{\circ}C$ but did 'IIOL $35^{\circ}C$. The cell size ofthe strain CS-2 was 2.9 p ~ n in length and 1.3 $\mu$ in diameter. Vegetable reproductmn was multiple budding and ascospre was present I to 4. Pseudomycelia or true myceliua formation were not observed In any of the cullureq. These results suggest that strain CS-2 is most likely a strain related Cryptococcus spp. (Cryptococcu spp. CS-2). When polygalacturonase or ihe yeast was induced by addition of polygalactoronic acid, polygalacturonase activity was detected in culture supernatent. There was a peak of specific activity a1 he mid-stationary phase(3 days culture) of growth. Polygalacturonase specific activity of Crylmcoccus sp. CS-2 was 2.96 unitsling. The molecular weighl ol'polygalacturonase was showed to be 46 KDa by both SDS-PAGE and activity stailling.

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Minority report; Diketopiperazines and Pyocyanin as Quorum Sensing Signals in Pseudomonas aeruginosa (Minority report; Pseudomonas aeruginosa의 정족수 인식(쿼럼 센싱) 신호물질로써의 Diketopiperazines과 Pyocyanin)

  • Lee, Joon-Hee
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.85-92
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    • 2008
  • Pseudomonas aeruginosa is an opportunistic human pathogen, causing a wide variety of infections including cystic fibrosis, microbial keratitis, and burn wound infections. The cell-to-cell signaling mechanism known as quorum sensing (QS) plays a key role in these infections and the QS systems of P. aeruginosa have been most intensively studied. While many literatures that introduce the QS systems of P. aeruginosa have mostly focused on two major acyl-homo serine lactone (acyl-HSL) QS signals, N-3-oxododecanoyl homoserine lactone (3OC12) and N-butanoyl homoserine lactone (C4), several new signal molecules have been discovered and suggested for their significant roles in signaling and virulence of P. aeruginosa. One of them is PQS (Pseudomonas quinolone signal; 2-heptyl-3-hydroxy-4-quinolone), which is now considered as a well-characterized major signal meolecule of P. aeruginosa. In addition, recent researches have also suggested some more putative signal molecules of P. aeruginosa, which are diketopiperazines (DKPs) and pyocyanin. DKPs are cyclic dipeptides and structurally diverse depending on what amino acids are involved in composition. Some DKPs from the culture supernatant of P. aeruginosa are suggested as new diffusible signal molecules, based on their ability to activate Vibrio fischeri LuxR biosensors that are previously considered specific for acyl-HSLs. Pyocyanin (1-hydroxy-5-methyl-phenazine), one of phenazine derivatives produced by P. aeruginosa is a characteristic blue-green pigment and redox-active compound. This has been recently suggested as a terminal signaling factor to upregulate some QS-controlled genes during stationary phase under the mediation of a transcription factor, SoxR. Here, details about these newly emerging signaling molecules of P. aeruginosa are discussed.