• Title/Summary/Keyword: co-substrate

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A novel cold-active lipase from Psychrobacter sp. ArcL13: gene identification, expression in E. coli, refolding, and characterization (새로운 Psychrobacter sp. ArcL13 유래 저온활성 지질분해효소 : 유전자 분리동정, 대장균에서의 발현, refolding 및 특성 연구)

  • Koo, Bon-Hun;Moon, Byung-Hern;Shin, Jong-Suh;Yim, Joung-Han
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.192-201
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    • 2016
  • Recently, Psychrobacter sp. ArcL13 strain showing the extracellular lipase activity was isolated from the Chuckchi Sea of the Arctic Ocean. However, due to the low expression levels of the enzyme in the natural strain, the production of recombinant lipase is crucial for various applications. Identification of the gene for the enzyme is prerequisite for the production of the recombinant protein. Therefore, in the present study, a novel lipase gene (ArcL13-Lip) was isolated from Psychrobacter sp. ArcL13 strain by gene prospecting using PCR, and its complete nucleotide sequence was determined. Sequence analysis showed that ArcL13-Lip has high amino acid sequence similarity to lipases from bacteria of some Psychrobacter genus (84-90%) despite low nucleotide sequence similarity. The lipase gene was cloned into the bacterial expression plasmid and expressed in E. coli. SDS-PAGE analysis of the cells showed that ArcL13-Lip was expressed as inclusion bodies with a molecular mass of about 35 kDa. Refolding was achieved by diluting the unfolded protein into refolding buffers containing various additives, and the highest refolding efficiency was seen in the glucose-containing buffer. Refolded ArcL13-Lip showed high hydrolytic activity toward p-nitrophenyl caprylate and p-nitrophenyl decanoate among different p-nitrophenyl esters. Recombinant ArcL13-Lip displayed maximal activity at $40^{\circ}C$ and pH 8.0 with p-nitrophenyl caprylate as a substrate. Activity assays performed at various temperatures showed that ArcL13-Lip is a cold-active lipase with about 40% and 73% of enzymatic activity at $10^{\circ}C$ and $20^{\circ}C$, respectively, compared to its maximal activity at $40^{\circ}C$.

Nutrient Dynamics in Decomposing Leaf Litter and Litter Production at the Long-Term Ecological Research Site in Mt. Gyebangsan (계방산 장기생태조사지의 낙엽 생산량 및 낙엽 분해에 따른 양분 동태)

  • Lee, Im-Kyun;Lim, Jong-Hwan;Kim, Choon-Sig;Kim, Young-Kul
    • Journal of Ecology and Environment
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    • v.29 no.6
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    • pp.585-591
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    • 2006
  • We measured the litterfall quantity and investigated the nutrient dynamics in decomposing litter for three years at the LTER sites installed in a deciduous broadleaf natural forest in Mt. Gyebangsan, South Korea. Litterfall production was significantly different among the sampling dates, whereas it was not significantly different among the years. The total annual mean litterfall production for three years was 6,593 kg $ha^{-1}$ $yr^{-1}$ and leaf litter accounted for 82.6% of the litterfall. The leaf litter quantity was highest in Quercus mongolia, followed by leaf of other species, Betula schmidtii, Kaplopanax pictus, Acer pseudo-sieboldianum, etc., which are dominant tree species in the site. The mass loss from the decomposition of leaf litter was fastest in Cortinus controversa (100%), followed by A. preudo-sieboldianum, K. pictus, and B. schmidtii. 100% of litter for C. controversa, 96.1% for A. pseudo-sieboldianum, 92.8% for K. pictus decomposed, while 66.2% of litter for Q. mongolia decayed for 1,003 days. The lower rate of the mass loss in the litter of Q. mongolia may be attributed to the difference in substrate quality, such as lower nutrient concentrations compared with those of other tree species. The concentrations of N, P, and Ca for five litter types increased over time, while the concentrations of K and Mg decreased over time. Compared with the nutrients in the litter of Q. mongolia, the nutrients (N, P, K, Ca, Mg) in the litter of other species, C. controversa, A. pseudo-sieboldianum, and K. pictus, were released more rapidly. The results showed that the mass loss and the nutrient dynamics in the litter are variable depending on the tree species even in the same site conditions.

Rumen Fermentation was Changed by Feed Inoculation Method in In Vitro (사료 접종 방법에 의한 in vitro 반추위 발효 상성 변화)

  • Yoo, Dae-Kyum;Moon, Joon-Beom;Kim, Han-Been;Yang, Sung-Jae;Park, Joong-Kook;Lee, Se-Young;Seo, Ja-Kyeom
    • ANNALS OF ANIMAL RESOURCE SCIENCES
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    • v.30 no.3
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    • pp.111-120
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    • 2019
  • The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

Analysis of the Effect of the Etching Process and Ion Injection Process in the Unit Process for the Development of High Voltage Power Semiconductor Devices (고전압 전력반도체 소자 개발을 위한 단위공정에서 식각공정과 이온주입공정의 영향 분석)

  • Gyu Cheol Choi;KyungBeom Kim;Bonghwan Kim;Jong Min Kim;SangMok Chang
    • Clean Technology
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    • v.29 no.4
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    • pp.255-261
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    • 2023
  • Power semiconductors are semiconductors used for power conversion, transformation, distribution, and control. Recently, the global demand for high-voltage power semiconductors is increasing across various industrial fields, and optimization research on high-voltage IGBT components is urgently needed in these industries. For high-voltage IGBT development, setting the resistance value of the wafer and optimizing key unit processes are major variables in the electrical characteristics of the finished chip. Furthermore, the securing process and optimization of the technology to support high breakdown voltage is also important. Etching is a process of transferring the pattern of the mask circuit in the photolithography process to the wafer and removing unnecessary parts at the bottom of the photoresist film. Ion implantation is a process of injecting impurities along with thermal diffusion technology into the wafer substrate during the semiconductor manufacturing process. This process helps achieve a certain conductivity. In this study, dry etching and wet etching were controlled during field ring etching, which is an important process for forming a ring structure that supports the 3.3 kV breakdown voltage of IGBT, in order to analyze four conditions and form a stable body junction depth to secure the breakdown voltage. The field ring ion implantation process was optimized based on the TEG design by dividing it into four conditions. The wet etching 1-step method was advantageous in terms of process and work efficiency, and the ring pattern ion implantation conditions showed a doping concentration of 9.0E13 and an energy of 120 keV. The p-ion implantation conditions were optimized at a doping concentration of 6.5E13 and an energy of 80 keV, and the p+ ion implantation conditions were optimized at a doping concentration of 3.0E15 and an energy of 160 keV.

Preparation and Gas Permeation Performance of Pd-Ag-Cu Hydrogen Separation Membrane Using α-Al2O3 Support (α-Al2O3 지지체를 이용한 Pd-Ag-Cu 수소 분리막의 제조 및 기체투과 성능)

  • Sung Woo Han;Min Chang Shin;Xuelong Zhuang;Jae Yeon Hwang;Min Young Ko;Si Eun Kim;Chang Hoon Jung;Jung Hoon Park
    • Membrane Journal
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    • v.34 no.1
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    • pp.50-57
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    • 2024
  • In this experiment, Pd-Ag-Cu membrane was manufactured using electroless plating on an α-Al2O3 support. Pd, Ag and Cu were each coated on the surface of the support through electroless plating and heat treatment was performed for 18 h at 500℃ in H2 in the middle of electroless plating to form Pd alloy. The surface of the Pd-Ag-Cu membrane was observed through Scanning Electron Microscopy (SEM), and the thickness of the Pd membrane was measured to be 7.82 ㎛ and the thickness of the Pd-Ag-Cu membrane was measured to be 3.54 ㎛. Energy dispersive X-ray spectroscopy and X-ray diffraction analysis confirmed the formation of a Pd-Ag-Cu alloy with a composition of Pd-78wt%, Ag-8.81wt% and Cu-13.19wt%. The gas permeation experiment was conducted under the conditions of 350~450℃ and 1~4 bar in H2 single gas and H2/N2 mixed gas. The maximum H2 flux of the hydrogen separation membrane measured in H2 single gas is 74.16 ml/cm2·min at 450℃ and 4 bar for the Pd membrane and 113.64 ml/cm2·min at 450℃ and 4 bar for the Pd-Ag-Cu membrane. In the case of the separation factor measured in H2/N2 mixed gas, separation factors of 2437 and 11032 were measured at 450℃ and 4 bar.