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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 $40^{\circ}C$.

Bioleaching of Mn(II) from Manganese Nodules by Bacillus sp. MR2 (Bacillus sp. MR2에 의한 망간단괴의 생물용출)

  • Choi, Sung-Chan;Lee, Ga-Hwa;Lee, Hong-Keum
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.411-415
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    • 2009
  • Some microorganisms are capable of leaching Mn(II) from nonsulfidic manganese ores indirectly via nonenzymatic processes. Such reductive dissolution requires organic substrates, such as glucose, sucrose, or galactose, as a source of carbon and energy for microbial growth. This study investigated characteristics of Mn(II) leaching from manganese nodules by using heterotrophic Bacillus sp. strain MR2 provided with corn starch as a less-expensive substrate. Leaching of Mn(II) at 25.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$ was accompanied with cell growth, but part of the produced Mn(II) re-adsorbed onto residual $MnO_2$ particles after 24 h. Direct contact of cells to manganese nodule was not necessary as a separation between them with a dialysis tube produced similar amount [24.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$]. These results indicated an involvement of extracellular diffusible compound(s) during Mn(II) leaching by strain MR2. In order to optimize a leaching process we tested factors that influence the reaction, and the most efficient conditions were $25\sim35^{\circ}C$, pH 5~7, inoculum density of 1.5~2.5% (v/v), pulp density of 2~3 g/L, and particle size <75 ${\mu}m$. Although Mn(II) leaching was enhanced as particle size decrease, we suggest <212 ${\mu}m$ as a proper size range since more grinding means more energy consumption The results would help for the improvement of bioleaching of manganese nodule as a less expensive, energy-efficient, and environment-friendly technology as compared to the existing physicochemical metal recovery technologies.

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.

Growth of CdSe thin films using Hot Wall Epitaxy method and their photoelectrical characteristics (HWE방법에 의한 CdSe 박막 성장과 광전기적 특성)

  • Hong, K.J.;Lee, K.K.;Lee, S.Y.;You, S.H.;Shin, Y.J.;Suh, S.S.;Jeong, J.W.;Jeong, K.A.;Shin, Y.J.;Jeong, T.S.;Kim, T.S.;Moon, J.D.;Kim, H.S.
    • Journal of Sensor Science and Technology
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    • v.6 no.4
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    • pp.328-336
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    • 1997
  • The CdSe thin films were grown on the Si(100) wafers by a hot wall epitaxy method (HWE). The source and substrate temperature are $600^{\circ}C$ and $430^{\circ}C$ respectively. The crystalline structure of epilayers was investigated by double crystal X-ray diffraction(DCXD). Hall effect on the sample was measured by the van der Pauw method and studied on the carrier density and mobility dependence on temperature. From Hall data, the mobility was increased in the temperature range 30K to 150K by impurity scattering and decreased in the temperature range 150k to 293k by the lattice scattering. In order to explore the applicability as a photoconductive cell, we measured the sensitivity(${\gamma}$), the ratio of photocurrent to darkcurrent(pc/dc), maximum allowable power dissipation(MAPD), spectral response and response time. The results indicated that the photoconductive characteristic were the best for the samples annealed in Cu vapor compare with in Cd, Se, air and vacuum vapour. Then we obtained the sensitivity of 0.99, the value of pc/dc of $1.39{\times}10^{7}$, the MAPD of 335mW, and the rise and decay time of 10ms and 9.5ms, respectively.

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Remediation of Heavy Metal Contaminated Groundwater by Using the Biocarrier with Dead Bacillus sp. B1 and Polysulfone (Bacillus sp. B1 사균과 Polysulfone으로 이루어진 미생물 담체를 이용한 중금속 오염 지하수 정화)

  • Lee, Min-Hee;Lee, Ji-Young;Wang, Soo-Kyun
    • Economic and Environmental Geology
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    • v.43 no.6
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    • pp.555-564
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    • 2010
  • Remediation process by using the bio-carrier (beads) with dead Bacillus sp. B1 and polysulfone was investigated for heavy metal contaminated groundwater. Sorption batch experiments using the bio-carrier were performed to quantify the heavy metal removal efficiencies from the contaminated solution. The analyses using SEM/EDS and TEM for the structure and the characteristic of precipitates on/inside the beads were also conducted to understand the sorption mechanism by the bio-carrier. Various amounts of freeze-dried dead Bacillus sp. B1 were mixed with polysulfone + DMF(N,N-dimethylformamide) solution to produce the bio-carrier (beads; less than 2mm in diameter) and 5% of Bacillus sp. B1 in the bio-carrier was optimal for Pb removal in the solution. The removal efficiency ratings of the bio-carrier for Pb, Cu and Cd were greater than 80% after adding 2g of bio-carrier in 50ml of aqueous solution (<10mg/L of each heavy metal concentration). Reaction time of the bio-carrier was very fast and most of the sorption reaction for heavy metals were completed within few hours. Batch experiments were duplicated at various pH conditions of aqueous solutions and Cu and Pb removal efficiencies highly maintained at wide pH ranges (pH 2-12), suggesting that the bio-carrier can be useful to clean up the acidic waste water such as AMD. From SEM/EDS and TEM analyses, it was observed that the bio-carrier was spherical shape and was overlapped by many porous layers. During the sorption experiment, Pb was crystallized on the surface of porous layers and also was mainly concentrated at the boundary of Bacillus sp. B1 stroma and polysulfone substrate, showing that the main mechanism of the bio-carrier to remove heavy metals is the sorption on/inside of the bio-carriers and the bio-carriers are excellent biosorbents for the removal of heavy metal ions from groundwater.

The Role of Factors Controlling the Accumulation of Biogenic Amines in Various Cheeses as Milk-Based Products: A Review (낙농유제품인 치즈에 축적된 생체 아민의 다양한 영향 인자에 관한 연구: 총설)

  • Chon, Jung-Whan;Kim, Dong-Hyeon;Kim, Hyun-Sook;Song, Kwang-Young;Lim, Jong-Soo;Choi, Dasom;Kim, Young-Ji;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.77-92
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    • 2014
  • Fermented foods have often been implicated as causative agents in poisoning due to toxic levels of biogenic amines. Cheese, a milk-based fermented food, is the product most likely to contain potentially harmful levels of biogenic amines, such as tyramine, histamine, putrescine, and so on. Recently, the risk awareness of a dietary uptake of high loads of biogenic amines has increased. Hence, we here review the published literature on several factors known to affect the biosynthesis of biogenic amines and their accumulation in milk-based foods. Furthermore, with regard to risk analysis, we discuss the control of factors related to the synthesis and accumulation of biogenic amines as a means to reduce their incidence in milk-based products, and thus to increase food safety.

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Effect of Water Activity and Temperature on Growth, Germination, Sporulation, and Utilization of Carbon Source of Penicillium oxalicum (PENOX) as a Biocontrol Agent(BCA) for control of Clover(Trifolium repens L.) (토끼풀(Trifolium repens L.) 방제용 생물제제 Penicillium oxalicum (PENOX)의 발아, 생장, 포자생성 및 탄소원이용에 미치는 수분활성 및 온도의 영향)

  • Lee, Hyang-Burm;Kim, Chang-Jin
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.68-74
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    • 2000
  • Penicillium oxalicum (PENOX) has shown the potential as a biocontrol agent(5CA) for control of a weed, clover(Trifolium repens L.) in grass plots. The bioherbicidal activity may be due to germinative and growth capacities and substrate availability of the agent over a range of environmental factors. The influences of different water activities($0.94{\sim}0.995\;a_w$) and temperatures($18{\sim}30^{\circ}C$) on mycelial growth, conidial germination, sporulation oil 2% MEA(malt extract agar) adjusted to different water activities with glycerol, and carbon source utilization using BIOLOG GN MicroPlate were determined in vitro. Decreases in $a_w$ on MEA caused a reduction in mycelial growth and conidial germination depending on temperature. The mycelial growth of PENOX was greatest at $30^{\circ}C/0.995\;a_w$. At some lowered water activity($0.97\;a_w$), the growth was similar between 25 and $30^{\circ}C$, and considerably decreased at lowered temperature($20^{\circ}C$). The germination rate was also greatest at $30^{\circ}C/0.995\;a_w$. Lag phase times for PENOX at $18^{\circ}C$ on MEA were >6hrs at tile whole $a_w$ level tested, and at 18 and $25^{\circ}C$ they were >18hrs and >12hrs at $0.94\;a_w$, respectively. However, its sporulation was some better at $0.97\;a_w$ than $0.995\;a_w$ or $0.94\;a_w$, and better at $20^{\circ}C$ than $30^{\circ}C$. In contrast, the number of carbon sources(niche size) utilized by PENOX varied with $a_w$ and temperature. Under some water stress condition($0.95\;a_w$), the agent utilized smaller number of carbon sources than $0.995\;a_w$ depending on temperature. The niche size at 0.995 and $0.95\;a_w$ were highest at $25^{\circ}C$, and showed 86 and 65, respectively. At $30^{\circ}C$, the niche size at 0.995 and $0.95\;a_w$ showed 84 and 50, respectively. There was no carbon source utilized by PENOX at $0.90\;a_w$ regardless of temperature. These information of tile fungal ecophysiology will be useful for the effective development of BCA.

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Three Phases and Water Characteristics of Horticultural Substrates (원예(園藝) 상토재료(床土材料)의 삼상(三相)과 수분특성(水分特性))

  • Jo, In-Sang;Hyun, Byung-Keun;Cho, Hyun-Jun;Jang, Yong-Seon
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.1
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    • pp.56-61
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    • 1997
  • A study was carried out to find out the basic information in physical properties for selection and manufacturing the good seedling media through the analysis of the physical properties, such as particle size, water retention and three phases of the major horticultural substrates. Easily available water(EAW), the water contents between 1kPa and 5kPa water potental, was highest in peatmoss with 39%, and perlite 34.0%, vermiculite 16.9%, but the values of osmunda and bark were lower than 4.8%. Water buffering capacity(WBC), the water content between 5kPa and 10kPa, was 6.1% in peatmoss and 2.3% in vermiculite but it was lower than 1.0% in other substrates. To adjust the suitable range of water potential at crossing point of water and air curves to 1.5~2.5kPa, more finer materials were needed in osmunda and bark, and more coarser materials must be added to peatmoss, perlite and vermiculite. Water potentials of substrates in saturated pot condition were equivalent to 2.2kPa in peatmoss and others were ranged in 1.0kPa to 4.3kPa of water potential in pressure chamber.

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Solar Module with a Glass Surface of AG (Anti-Glare) Structure (연요철(Anti-Glare) 구조의 표면 유리 기판을 가지는 고효율 태양전지 모듈)

  • Kong, Dae-Young;Kim, Dong-Hyun;Yun, Sung-Ho;Bae, Young-Ho;Yu, In-Sik;Cho, Chan-Seob;Lee, Jong-Hyun
    • Journal of the Korean Vacuum Society
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    • v.20 no.3
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    • pp.233-241
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    • 2011
  • Currently, solar module is using the two methods such as a glass-filled method or a super-straight method. The common point of these methods is to use glass structure on the front of solar module. However, the reflectance of the solar module is high depending on the height of the incident sunlight due to the flat surface of the module front glass. Purposed to solve these problems, AG (anti-glare) structures were formed on the glass surface. Next is fabrication methods of AG structure. First, uneven structure made by micro blaster equipment was dipped in Hydro-fluidic acid (HF) acid. HF acid process was carried out to remove particles and to make high transmittance. The reflectance and transmittance of the anti-glare glass was compared to those of the bare glass. The reflectance of anti-glare glass decreased approximately 1% compared with bare glass. The transmittance of anti-glare glass was similar to bare glass. According to the sample angle, the difference of the reflectance between bare glass and the anti-glare glass was about 19%. Isc and efficiency value of anti-glare glass on bare solar cell appeared about 3.01 mA and 0.228% difference compared with bare glass. Anti-glare glass on textured solar cell appeared about 9.46 mA and 0.741% difference compared with bare glass. As a result, the role of anti-glare in the substrate is to reduces the loss of sunlight reflected from the surface. In this study, therefore, AG structure on the solar cell was used to improve the efficiency of solar cell.

Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents (세탁세제 첨가용 효소 개발을 위한 남극 해양세균 유래 저온성 단백질분해효소의 특성 연구)

  • Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.60-68
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    • 2018
  • A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.