• Title/Summary/Keyword: 배양액 농도

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Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.

유리화 동결법에 의한 난자의 동결 중 항동해제에 Cytoskeleton Stabilizer, Taxol의 처리 후 배발달율의 개선에 관한 연구

  • 정형민;박성은;임정묵;고정재;정미경;차광렬
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.74-74
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    • 2001
  • 난자 동결방법의 선별은 보다 효과적인 난자은행의 개발에 필수 불가결한 중요한 요소이다. 이전의 연구에서 마우스의 난자를 ethylene glycol과 electron microscope grid를 이용한 유리화 동결법으로 동결 융해한 결과 기존의 slow freezing 방법에서보다 높은 생존율과 배발달율이 나타남을 관찰하였다. 그러나 동결융해후의 난자는 방추사와 염색체의 이상성이 대조군에 비해 높은 빈도로 나타나 융해후의 배발달율을 감소시키는 것으로 보고되었다. 이에 본 연구에서는 유리화동결법동안 항동해제에 Cytoskeleton system을 안정화시키는 cytoskeleton stabilizer인 taxol을 첨가시킨후 동결시켰을때 생존율과 발달율을 개선시킬 수 있는지 알아보고자 본 실험을 시행하였다. ICR mouse의 성숙란을 채취하여 연구목적에 따라 taxol을 첨가시키지 않은 대조군과 첨가시킨 실험군으로 분류하였다. 동결방법은 난자를 1.5 M ethylene glycol (EG)에 2분 30초간 노출시킨후 5.5 M EG와 1 M sucrose가 첨가된 동결액에 20초간 노출시킨 후 Grid에 난자를 부착시킨후 직접 액체질소에 침지하여 동결하였다. 동결후 난자는 5단계로 융해를 실시한 후 정자와 체외수정을 시킨 후 수정된 난자는 modified P1 배약액에 124 h까지 발달율을 관찰하였고, 배양 후 발달된 배반포는 대조군과 실험군, 각각 4마리의 발정동기화된 recipient에 이식을 시행하였다. 배발달율은 대조군에 비해 실험군에서 4세포기 (48 vs. 84.4%), 8세포기 (34% vs. 70.6%), 상실배 (26% vs. 58.6%) 그리고 배반포 발달율은 (24% vs. 58.6%)로 높게 관찰되었다. 배아이식후 대조군과 실험군에서 각각 2 마리가 임신이되어 정상적인 산자를 분만하였다. 따라서 항동해제에 taxol의 첨가는 동결 융해후의 난자의 배발달율을 증진시킬 수 있었다..8%로 나타나 난할율 및 배반포 발생율에 있어서 융합조건에 따라 큰 차이는 없었으나 1.9㎸/cm, 30$\mu\textrm{s}$ 2회의 조건이 다른 조건들에 비하여 유의적으로 낮았다. 따라서, 체세포와 수핵란 세포질간의 융합율과 배반포 발생에 미치는 영향은 전압보다는 시간에 더 크게 받음을 알 수 있었으며, 이와 같은 결과에서 융합시 시간을 오래 주는 것보다 전압을 높이는 것이 수핵난자의 세포질에 상해를 줄이고 이후 배반포 발생에 유리할 것으로 사료되었다.면에서도 더욱 더 활발할 것으로 기대된다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며 큰 자기폭풍일수록 현저했다. 주상에서 관측된 이러한 특성은 서브스톰 확장기 활동이 자기폭풍의 발달과 밀접한 관계가 있음을 시사한다.se that were all low in two aspects, named "the Nonsignificant group". And the issues were high risk perception in general setting and

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Continuous Hydrogen Production by Heterotrophic Growth of Citrobacter amalonaticus Y19 in Trickle Bed Reactor (Citrobacter amalonaticus Y19의 영양종속 성장을 이용한 Trickle Bed Reactor에서의 연속적인 수소생산)

  • Park, Ji-Young;Lee, Tae-Ho;Oh, You-Kwan;Kim, Jun-Rae;Seol, Eun-Hee;Jung, Gyoo-Yeol;Kim, Mi-Sun;Park, Sung-Hoon
    • KSBB Journal
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    • v.20 no.6
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    • pp.458-463
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    • 2005
  • [ $H_2$ ] from CO and water was continuously produced in a trickle bed reactor(TBR) using Citrobacter amalonaticus Y19. When the strain C. was cultivated in a stirred-tank reactor under a chemoheterotrophic and aerobic condition, the high final cell concentration of 13 g/L was obtained at 10 hr. When the culture was switched to an anaerobic condition with the continuous supply of gaseous CO, CO-dependent hydrogenase was fully induced and its hydrogen production activity approached 16 mmol/g cell/hr in 60 hr. The fully induced C. amalonaticus Y19 cells were circulated through a TBR packed with polyurethane foam, and the TBR was operated for more than 20 days for $H_2$ production. As gas retention time decreased or inlet CO partial pressure increased, $H_2$ production rate increased but the conversion from CO to $H_2$ decreased. The maximum $H_2$ production rate obtained was 16 mmol/L/hr at the gas retention time of 25 min and the CO inlet partial pressure of 0.4 atm. The high $H_2$ production rate was attributed to the high cell density in the liquid phase circulating the TBR as well as the high surface area of polyurethane foam used as packing material of the TBR.

Optimization of Bisphenol A Biodegradation by Trametes versicolor (Trametes versicolor에 의한 Bisphenol A 생분해의 최적조건)

  • Kang, Ae-Ri;Choi, Hyoung-Tae;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.37-42
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    • 2008
  • Optimal conditions for the biodegradation of endocrine-disrupting bisphenol A (BPA) were examined for the white rot fungus Trametes versicolor isolated in Korea. T. versicolor degraded 100% of 50 mg/L bisphenol A during 12 hr in yeast extract-malt extract-glucose (YMG) medium. When BPA was added to the 5-day preincubated fungal culture in YMG medium, all BPA was removed in 2 hr. T. versicolor could efficiently degrade BPA at $35^{\circ}C$, pH 6 in YMG medium. T. versicolor could more easily remove BPA of $1{\sim}25\;mg/L$ than that of higher concentrations ($50{\sim}100\;mg/L$) in YMG medium. T. versicolor degraded 100% of 50 mg/L BPA for 36 h in a minimal medium, which is lower degradation rate than that in YMG medium. Optimal conditions for BPA biodegradation in the minimal medium were similar to those in YMG medium. When BPA (50 mg/L) was added into domestic wastewater, it could be completely removed by T. versicolor. During the biodegradation of BPA by T. versicolor in YMG medium, its estrogenic activity decreased.

Comparative Analysis of Resistance to Antibiotics in Populus alba$\times$P. glandulosa Transformed by nptII or hpt Gene (NPTII 및 HPT 유전자가 삽입된 현사시의 항생제에 대한 저항성 분석)

  • 이은정;노은운;박재인
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.243-248
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    • 2001
  • This study was peformed to find out the optimal conditions for the selection of transformed cells using already established transgenic plants. Several transgenic poplar (Populus alba$\times$P giandulosa) lines carrying npt II or hpt gene as a selectable marker were tested against kanamycin or hygromycin. Two culture explants, leaf discs and nodes, were compared regarding their sensitivity to the antibiotics. When leaf discs of untransformed control plants were cultured on callus inducing media in the presence of varying levels of kanamycin or hygromycin, only those cultured on the media containing lower than 50 mg/L kanamycin or 2 mg/L hygromycin formed callus. However, much higher concentration of kanamycin was needed to suppress the growth of axillary buds of untransformed plants. On the other hand, hygromycin at the concentration of 5 mg/L effectively suppressed shoot growth of untransformed plants. Root induction from untransformed plants could also be suppressed at the concentration of 50 mg/L kanamycin or 5 mg/L hygromycin. The transgenic plants showed resistance to 100 mg/L kanamycin or 50 mg/L hygromycin in the growth of callus, shoots, and roots. Hygromycin appeared to be more efficient in selecting untransformed cells than kanamycin.

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Isolation of Bacillus velezensis SSH100-10 with Antifungal Activity from Korean Traditional Soysauce and Characterization of Its Antifungal Compounds (전통재래 간장으로부터 항진균 활성 B. velezensis SSH100-10의 분리와 그 항진균 물질의 특성 구명)

  • Chang, Mi;Moon, Song Hee;Chang, Hae Choon
    • Food Science and Preservation
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    • v.19 no.5
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    • pp.757-766
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    • 2012
  • The SSH100-10 bacterial strain, which exhibits strong antifungal (anti-mold and anti-yeast) activity, was isolated from traditional korean soysauce aged 100 years. The strain was identified as Bacillus velezensis based on Gram-staining, the biochemical properties and 16S rRNA gene sequence determination. B. velezensis SSH100-10 showed strong proteinase activity and NaCl tolerance, but did not produce enterotoxin. Two-antifungal compounds from B. velezensis SSH100-10 were purified using SPE, preparative HPLC, and reverse phase-HPLC. The purified antifungal compounds were identified as $C_{14}$ and $C_{15}$ iturin through MALDI-TOF-MS and amino acid composition analysis. The stability characteristics of the antifungal compounds after temperature, pH, and enzyme treatments suggested that B. velezensis SSH100-10 produced more than two antifungal compounds; pH-stable $C_{14}$ iturin A and $C_{15}$ iturin A, and unidentified pH-unstable compounds. The results suggested that B. velezensis SSH100-10 can be used in soybean fermentation as a starter. Moreover it has potential as a biopreservative in the food and feed industry and as a biocontrol agent in the field of agriculture.

Some Factors Affecting the Protoplast Formation and Regeneration from the Mycelium of Ganoderma lucidum(Fr.) Karsten (영지(靈芝)버섯 균사(菌絲)의 원형질체(原形質體) 나출(裸出) 및 재생(再生))

  • Shin, Gwan Chull;Yeo, Un Hyung;Yoo, Young Bok;Park, Yong Hwan
    • Korean Journal of Agricultural Science
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    • v.13 no.2
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    • pp.185-192
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    • 1986
  • Some factors affecting the protoplast release from mycelia of Ganoderma lucidum and regeneration of the protoplast were investigated and the results obtained are summarized as follows; Novozym 234 as a lytic enzyme was the most effective for the protoplast release from mycelia of Ganoderma lucidu m and its optimal concentration was 10mg per ml of osmotic stabilizer. The highest number of protoplasts were released after 3 hours incubation in the reciprocal shaking bath at 120 oscillations a minute. Among six osmotic stabilizers tested, 0.6M sucrose showed the best result. SCM medium showed good mycelial growth and high yields of protoplasts. The protoplasts released from the mycelium of G. lucidum were regenerated at 0.20 to 0.27 percent on MCM, MMM and SCM. Of the cultures obtained from protoplasts regenerated, 13 to 29 percent were monokaryon.

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Evaluation of Properties of Sulfur-Oxidizing Bacteria Growth and Resistance to Biochemical Corrosion by Simulation Test (시뮬레이션 시험에 의한 황산화세균의 생장 특성 및 생화학적 부식 저항성 평가)

  • Kim, Gyu-Yong;Lee, Eui-Bae;Khil, Bae-Su;Lee, Seung-Hun
    • Journal of the Korea Concrete Institute
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    • v.20 no.4
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    • pp.495-502
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    • 2008
  • To analyze the growth of SOB(Thiobacillus novellus) and biochemical corrosion of concrete, simulation test method and device were developed, and basic conditions for SOB growth were established. Two types of simulation tests were conducted according to a transplant method and a concentration of $H_2SO_4$. As a result, the SOB growth in distinct manners and antibiosis of specimen were observed. In the case of the specimens indirectly transplanted with SOB through culture solution submersion at a hydrogen sulfide level of 120 ppm, the rapid activation of SOB and the resulting sulfuric acid production were observed. However, SOB were shown to grow rapidly and then die out in a relative short period of time. Meanwhile, in the case of the specimens directly transplanted with SOB at a hydrogen sulfide level of 50 ppm, the long-term growth of SOB was possible, but the production of sulfuric acid by SOB did not progress. In the case of the antibiotic metal-mixed specimens, SOB with destroyed cell membranes and internal organizations were observed.

Characterization and Purification of Subtilosin A Produced by Bacillus vallismortis MCBL 1012 Isolated from Seasoned Dried Radish

  • Se-Yeon Lee;Dae-Ook Kang
    • Journal of Life Science
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    • v.34 no.8
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    • pp.576-587
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    • 2024
  • In this study, diverse bacterial strains were isolated from fermented foods to screen those with antibacterial activity. Among them, one strain, identified as Bacillus vallismortis MCBL 1012 through 16S rRNA gene sequence analysis, was selected for its bacteriocin production. The culture supernatant of B. vallismortis MCBL 1012 showed antibacterial activity, mainly against Gram-positive bacteria. Scanning electron microscopy (SEM) revealed that bacteriocin treatment led to cellular content leakages in Listeria monocytogenes KCCM 40307, Enterococcus faecium KCCM 12118, and Streptococcus mutans KCTC 3065. PCR analysis confirmed B. vallismortis MCBL 1012 harbored subtilosin A gene (sbo A). Antibacterial activity was decreased by proteolytic enzymes like proteinase K, subtilisin A, and α-chymotrypsin. The bacteriocin demonstrated stability at 40℃ and 60℃ for 120 min, and up to 80℃ for 60 min, with rapid activity loss at 100℃. It retained full antibacterial activity within a pH range of 4.0 to 8.0 and was not affected by up to 100% organic solvents like ethanol, methanol, acetonitrile, and tetrahydrofuran. Nevertheless, activity decreased with more than 40% isopropanol and 80% acetone. Most tested inorganic salts and detergents had no effect on antibacterial activity except, CuSO4 and NiSO4 at specified concentrations. The bacteriocin exerted its antibacterial effect through bactericidal action against L. monocytogenes KCCM 40307. The bacteriocin was purified by ammonium sulfate precipitation, DEAE anion exchange chromatography, and RP-HPLC. The purification resulted in a final yield of 0.03% and a 283.7-fold increase in specific activity. MALDI-TOF MS analysis determined the exact molecular weight of purified bacteriocin to be 3,326.1 Da.

The Effect of Surfactant on Neutrophil Apoptosis in Lipopolysaccharide Induced Acute Lung Injury in Rat (기관내 내독소 투여로 유도한 백서의 급성 폐손상 모델에서 surfactant가 호중구의 아포토시스에 미치는 영향)

  • Yoo, Ji-Hoon;Lee, Byoung-Jun;Jeong, Do-Young;Lee, Sang-Hoon;Shin, Jong-Wook;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung-Whui
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.4
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    • pp.409-419
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    • 2002
  • Background : The therapeutic effects of surfactant on acute lung injury derive not only from its recruiting action on collapsed alveoli but also from its anti-inflammatory effects. Pro-apoptotic action on alveolar neutrophils represents one of the important anti-inflammatory mechanisms of surfactant. In the present study, we evaluated the effects of sufactant on the apoptosis of human peripheral and rat alveolar neutrophils. Methods : In the (Ed- the article is not definitely needed but it helps to separate the two prepositions 'in') in vitro study, human neutrophils were collected from healthy volunteers. An equal number of neutrophils ($1{\times}10^6$) (Ed-confirm) was treated with LPS (10, 100, 1000ng/ml), surfactant (10, 100, $1000{\mu}g/ml$), or a combination of LPS (1000ng/ml) and surfactant (10, 100, $1000{\mu}g/ml$). After incubation for 24 hours, the apoptosis of neutrophils was evaluated by Annexin V method. In the in vivo study, induction of acute lung injury in SD rats by intra-tracheal instillation of LPS (5mg/kg) was followed by intra-tracheal administration of either surfactant (30mg/kg) or normal saline (5ml/kg). Tenty-four hours after LPS instillation, alveolar neutrophils were collected and the apoptotic rate was evaluated by Annexin V method. In addition, changes of the respiratory mechanics of rats (respiratory rate, tidal volume, and airway resistance) were evaluated with one chamber body plethysmography before, and 23 hours after, LPS instillation. Results : in the in vitro study, LPS treatment decreased the apoptosis of human peripheral blood neutrophils (control: $47.4{\pm}5.0%$, LPS 10ng/ml; $30.6{\pm}10.8%$, LPS 100ng/ml; $27.5{\pm}9.5%$, LPS 1000ng/ml; $24.4{\pm}7.7%$). The combination of low to moderate doses of surfactant with LPS promoted apoptosis (LPS 1000ng/ml + Surf $10{\mu}g/ml$; $36.6{\pm}11.3%$, LPS 1000ng/ml +Surf $100{\mu}g/ml$; $41.3{\pm}11.2%$). The high dose of surfactant ($1000{\mu}g/ml$) decreased apoptosis ($24.4{\pm}7.7%$) and augmented the anti-apoptotic effect of LPS (LPS 1000ng/ml + Surf $1000P{\mu}g/ml$; $19.8{\pm}5.4%$). In the in vivo study, the apoptotic rate of alveolar neutrophils of surfactant-treated rats was higher than that of normal saline-treated rats ($6.03{\pm}3.36%$ vs. $2.95{\pm}0.58%$). The airway resistance (represented by Penh) of surfactant-treated rats was lower than that of normal saline-treated rats at 23 hours after LPS injury ($2.64{\pm}0.69$ vs. $4.51{\pm}2.24$, p<0.05). Conclusion : Surfactant promotes the apoptosis of human peripheral blood and rat alveolar neutrophils. Pro-apoptotic action on neutrophils represents one of the important anti-inflammatory mechanisms of surfactant.