• 제목/요약/키워드: High temperature incubation

검색결과 216건 처리시간 0.028초

Mouse 배의 Glass화 보존 (Cryopreservation of Mouse Embryos by Vitrification)

  • 권오용
    • 한국가축번식학회지
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    • 제13권2호
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    • pp.63-69
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    • 1989
  • Eight-cell mouse embryos equilibrated with vitrification solution(VS3), consisting of glycerol and polyethlene glycol, were plunged directly into liquid nitrogen. The embryos were cryopreserved by vitrification without intra- and extracellular ice formation. After the vitrified embryos were warmed at 4$^{\circ}C$, sucrose dilution procedures were examined and the survival embryos were transferred to recipients after 48h incubation in vitro. The results were obtained as follows. 1. Mouse embryos equilibrated with VS3 were diluted with 1.5m sucrose-HB 1 solution, 0.5M sucrose-HB1 solution for 5min at room temperature, respectively. The proportion of vitrified-warmed embryos developed to blastocyst(85.7%) was as high as that of the embryos diluted with 1.04M sucrose-HB1 solution at 4$^{\circ}C$(85.5%). 2. Normal live youngs were obtained in 53.9%(55/102) of the vitrified-warmed embryos after tranfer to pseudopregnant recipients.

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Studies on Microbial Transformation of Meloxicam by Fungi

  • Shyam Prasad, G.;Girisham, S.;Reddy, S.M.
    • Journal of Microbiology and Biotechnology
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    • 제19권9호
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    • pp.922-931
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    • 2009
  • Screening-scale studies were performed with 26 fungal cultures for their ability to transform the anti-inflammatory drug meloxicam. Among the different fungi screened, a filamentous fungus, Cunninghamella blakesleeana NCIM 687, transformed meloxicam to three metabolites in significant quantities. The transformation of meloxicam was confirmed by high-performance liquid chromatography (HPLC). Based on the liquid chromatography-tandem mass spectrometry (LC-MS/MS) data, two metabolites were predicted to be 5-hydroxymethyl meloxicam and 5-carboxy meloxicam, the major mammalian metabolites reported previously. A new metabolite was produced, which is not detected in mammalian systems. Glucose medium, pH of 6.0, temperature of $27^{\circ}C$, 5-day incubation period, dimethylformamide as solvent, and glucose concentration of 2.0% were found to be suitable for maximum transformation of meloxicam when studied separately. It is concluded that C. blakesleeana can be employed for biotransformation of drugs for production of novel metabolites.

Enzymatic Properties of Protease from the Hepatopancreas of Shrimp, Penaeus japonicus

  • Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • 제3권3_4호
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    • pp.188-194
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    • 2000
  • A protease purified from hepatopancreas of shrimp, Penaeus japonicus, had maximum activity at $70^{\circ}C$ and in neutral and alkaline pH ranges. Specific activity at optimum reaction condition of the protease was estimated to be approximately 12 U/mg/min. The protease was stable in neutral and alkaline pH ranges and activity was retained after heat treatment at $50^{\circ}C$ for 30 min. Apparent $K_m$ and $V_{max}$ value against casein substrate were estimated to be $0.29\%$ and $7.8see^{-1}$, respectively, and those against N-CBZ-L-tyrosine p-nitropheny1 ester (CBZ­Tyr-NE) were 0.38 mM and $2,400 see^{-1}$, respectively. The N-termina1 sequence of the protease showed high homology to the trypsin from same species and the proteases from shrimp. Myosin heavy chain (MHC) from shrimp tail meat was the most susceptible to the protease and actin/tropomyosin were degraded progressively during 4 hr incubation, but to a lesser degree than MHC.

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Flammulina velutipes에 의한 Laccase의 생산과 효소적 특성 (Production and Enzymatic Properties of Laccase from Flammulina velutipes)

  • 이재성;서달선
    • 한국균학회지
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    • 제13권2호
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    • pp.111-114
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    • 1985
  • The production of lac case by the funguson various media was studied. The characteristics of the enzyme were also studied regarding to the optimum pH, stability, Km value, and inactivation. The maximum activity of laccase reached the 40 days of incubation and the barley straw extract appeared to be a strong inducer for laccase. The enzyme showed stability at wide range of pH with optimum pH of 6.6. Temperature stability of the enzyme was high. Laccase was not inactivated by the organic solvents used for the precipitation. The enzyme, how­ever, was completely inactivated by trichloroacetic acid and sodium azide.

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IoT 기반의 스마트 팜 시스템 구조설계에 관한 연구 (A Study on the Architecture Design of Smart Farm System based on IoT Technology)

  • 길민식;곽동걸;최신형;신종근
    • 전력전자학회:학술대회논문집
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    • 전력전자학회 2019년도 전력전자학술대회
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    • pp.543-545
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    • 2019
  • Recently, the demand for smart farms is increasing due to the increase in the cultivation area such as horticulture, fruit trees and special crops. However, due to the irregular weather changes and the cultivation method of the crops due to the different cultivation environment, there are frequent occurrence of diseases and insect pests and infectious diseases due to system error or carelessness, and the cycle is also very short. In addition, the Smart Farm business has been built by combining various sensors (temperature, humidity, CO2, illumination) and LED lighting, but it is costly in terms of frequent errors, lack of power supply, And thus the management can not be efficiently managed. Therefore, this paper combines real time sensing technology based on IoT Platform and high performance control technology to control pests and equipment errors and monitor the growth status of crops in real time based on big data analysis and Artificial Intelligence System.

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Novel Low-Temperature-Active Phytase from Erwinia carotovora var. carotovota ACCC 10276

  • Huang, Huoqing;Luo, Huiying;Wang, Yaru;Fu, Dawei;Shao, Na;Yang, Peilong;Meng, Kun;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • 제19권10호
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    • pp.1085-1091
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    • 2009
  • A phytase with high activity at low temperatures has great potential for feed applications, especially in aquaculture. Therefore, this study used a degenerate PCR and TAIL PCR to clone a phytase gene from Erwinia carotovora var. carotovota, the cause of soft rot of vegetables in the ground or during cold storage. The full-length 2.5-kb fragment included an open reading frame of 1,302 bp and encoded a putative phytase of 45.3 kDa with a 50% amino acid identity to the Klebsiella pneumoniae phytase. The phytase contained the active site RHGXRXP and HD sequence motifs that are typical of histidine acid phosphatases. The enzyme was expressed in Escherichia coli, purified, and displayed the following characteristics: a high catalytic activity at low temperatures (retaining over 24% activity at $5^{\circ}C$) and remarkably thermal lability (losing >96% activity after incubation at $60^{\circ}C$ for 2 min). The optimal phytase activity occurred at pH 5.5 and ${\sim}49^{\circ}C$, and the enzyme activity rapidly decreased above $40^{\circ}C$. When compared with mesophilic counterparts, the phytase not only exhibited a high activity at a low temperature, but also had a low $K_m$ and high $k_{cat}$. These temperature characteristics and kinetic parameters are consistent with low-temperature-active enzymes. To our knowledge, this would appear to be the first report of a low-temperature-active phytase and its heterogeneous expression.

High-Level Expression of Pseudomonas sp. LBC505 Endoglucanase Gene in Escherichia coli

  • Chun, Sung-Sik;Kim, Yang-Woo;Chung, Young-Chul;Kim, Kyeong-Sook;Sung, Nack-Kie
    • Journal of Microbiology and Biotechnology
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    • 제5권1호
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    • pp.14-17
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    • 1995
  • Endoglucanase gene of Pseudomonas sp. LBC505 was previously cloned in pUC19 to yield plasmid pLCl. The Pseudomonas sp. LBC505 endoglucanase gene was subcloned in a temperature-regulated Es-cherichia coli expression vector, pAS1, containing the leftward promoter $P_L$ of bacteriophage lambda. The level of gene expression was controlled by the thermal inactivation of the heat-sensitive lambda cI857 repressor. Best yield of endoglucanase was obtained by lowering the incubation temperature to $37^{\circ}C$ after induction at $42^{\circ}C$ for 1h. Under these conditions enzyme production continued for about 5h at a gradually decreasing rate. Ecoli harboring recombinant plasmid pASC10 expressed 4.3 times as much CMCase activity as E.coli containing pLCl. To enhance the expression level of endogl, ucanase gene, we have also changed the presumptive Shine-Dalgamo sequence (AGAGGT) of the gene to consensus sequence (AGGAGGT) by site-directed mutagenesis. The genes mutated were subcloned in pASl resulting in the formation of recombinant plasmid pASS50. E.coli harboring the plasmid pASS50 expressed 6.2-fold higher levels of CMCase activity than that of E.coli harboring pLC1.

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지열시스템의 그라우트 및 수온변화가 미생물에 미치는 영향 실험 (Effects of Grouts and Temperature Change on Microorganisms in Geothermal Heat Pump)

  • 조윤주;이진용;김창균;한지선
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제14권4호
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    • pp.10-14
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    • 2009
  • 본 논문은 지열시스템의 설치 시 사용되는 그라우트와 운영에 따른 수온변동이 유발할 수 있는 미생물학적 영향을 실내실험을 통해 살펴보았다. 시료는 지열히트펌프가 아직 가동되지 않는 관정(한방병원)과 지열 히트펌프가 가동 중인 곳(창업보육센터)에서 채취하였다. 실험에 사용한 그라우트는 볼클레이 벤토나이트로 나트륨(Na)계이며 Real-time PCR을 사용하여 각 시료에서 추출된 총 유전자 DNA 중 세균이 가지고 있는 16S rDNA를 정량함으로써 전체 세균의 양을 평가하였다. 실험 결과 지열히트펌프가 가동 중인 지하수에서 총 세균양이 가장 많았으며 벤토나이트를 주입하면서 세균수가 증가하는 경향을 보였다. 한편 그라우트 및 수온영향을 보다 명확하게 파악하기 위해서는 장기적인 현장모니터링이 요구된다.

Immobilization of GH78 α-L-Rhamnosidase from Thermotoga petrophilea with High-Temperature-Resistant Magnetic Particles Fe3O4-SiO2-NH2-Cellu-ZIF8 and Its Application in the Production of Prunin Form Naringin

  • Xu, Jin;Shi, Xuejia;Zhang, Xiaomeng;Wang, Zhenzhong;Xiao, Wei;Zhao, Linguo
    • Journal of Microbiology and Biotechnology
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    • 제31권3호
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    • pp.419-428
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    • 2021
  • To efficiently recycle GH78 thermostable rhamnosidase (TpeRha) and easily separate it from the reaction mixture and furtherly improve the enzyme properties, the magnetic particle Fe3O4-SiO2-NH2-Cellu-ZIF8 (FSNcZ8) was prepared by modifying Fe3O4-NH2 with tetraethyl silicate (TEOS), microcrystalline cellulose and zinc nitrate hexahydrate. FSNcZ8 displayed better magnetic stability and higher-temperature stability than unmodified Fe3O4-NH2 (FN), and it was used to adsorb and immobilize TpeRha from Thermotoga petrophilea 13995. As for properties, FSNcZ8-TpeRha showed optimal reaction temperature and pH of 90℃ and 5.0, while its highest activity approached 714 U/g. In addition, FSNcZ8-TpeRha had better higher-temperature stability than FN. After incubation at 80℃ for 3 h, the residual enzyme activities of FSNcZ8-TpeRha, FN-TpeRha and free enzyme were 93.5%, 63.32%, and 62.77%, respectively. The organic solvent tolerance and the monosaccharides tolerance of FSNcZ8-TpeRha, compared with free TpeRha, were greatly improved. Using naringin (1 mmol/l) as the substrate, the optimal conversion conditions were as follows: FSNcZ8-TpeRha concentration was 6 U/ml; induction temperature was 80℃; the pH was 5.5; induction time was 30 min, and the yield of products was the same as free enzyme. After repeating the reaction 10 times, the conversion of naringin remained above 80%, showing great improvement of the catalytic efficiency and repeated utilization of the immobilized α-L-rhamnosidase.

Supplementation of Essential Oil Extracted from Citrus Peel to Animal Feeds Decreases Microbial Activity and Aflatoxin Contamination without Disrupting In vitro Ruminal Fermentation

  • Nam, I.S.;Garnsworthy, P.C.;Ahn, Jong Ho
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권11호
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    • pp.1617-1622
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    • 2006
  • Long-term storage of feeds or feedstuffs in high temperature and humid conditions can be difficult because of microbial contamination. Essential oil isolated from industrial waste citrus peel could be used as a preservative because it is likely to have anti-bacterial and anti-fungal activity. Our objective was to determine whether different levels (0.028, 0.056 and 0.112 g/kg) of citrus essential oil (CEO) would provide anti-microbial activity and enhance preservation of animal feed without influencing rumen fermentation. At 0.112 g/kg, CEO inhibited growth of Escherichia coli (ATCC 25922) and Salmonela enteritidis (IFO 3313). Growth of E. coli recovered after 24 h of incubation, but S. enteritidis continued to be inhibited for 72 h. Preservation of antibiotic-free diets for swine was assessed by observing anti-aflatoxin activity. Aflatoxin was detected in control feed samples on days 16 (8 ppb) and 21 (8 ppb) and in anti-fungal agent (AA) treated samples on days 16 (2 ppb) and 21 (4 ppb). However, aflatoxin was not detected in feed samples treated with CEO. Treatment with CEO and AA did not influence ruminal pH, dry matter digestibility (DMD) or organic matter digestibility (OMD) over 48 h of incubation in rumen fluid. Acetate and propionate were slightly higher with CEO treatment (p<0.05), but total concentration of volatile fatty acid (VFA) was not significantly affected by treatment. Ammonia-N concentration was slightly higher for the control treatment (p<0.05). This study showed that treating feed with CEO enhances preservation of animal feed without influencing in vitro rumen fermentation.