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A Direct Approach for Finding Functional Lipolytic Enzymes from the Paenibacillus polymyxa Genome

  • JUNG, YEO-JIN;KIM, HYUNG-KWOUN;KIM, JIHYUN F.;PARK, SEUNG-HWAN;OH, TAE-KWANG;LEE, JUNG-KEE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.155-160
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    • 2005
  • Abstract A direct approach was used to retrieve active lipases from Paenibacillus polymyxa genome databases. Twelve putative lipase genes were tested using a typical lipase sequence rule built on the basis of a consensus sequence of a catalytic triad and oxyanion hole. Among them, six genes satisfied the sequence rule and had similarity (about 25%) with known bacterial lipases. To obtain the six lipase proteins, lipase genes were expressed in E. coli cells and lipolytic activities were measured by using tributyrin plate and pnitrophenyl caproate. One of them, contig 160-26, was expressed as a soluble and active form in E. coli cell. After purifying on Ni-NTA column, its detailed biochemical properties were characterized. It had a maximum hydrolytic activity at $30^{\circ}C$ and pH 7- 8, and was stable up to $40^{\circ}C$ and in the range of pH 5- 8. It most rapidly hydrolyzed pNPC$_6$ among various PNPesters. The other contigs were expressed more or less as soluble forms, although no lipolytic activities were detected. As they have many conserved regions with lipase 160-26 as well as other bacterial lipases throughout their equence, they are suggested as true lipase genes.

A Study on Bending Fatigue Strength of One Side Fillet Welded T-Joint by SM 490A steel (Sm 490A강으로 제작된 T형 편면용접이음재의 굽힘피로강동에 관한 연구)

  • 엄동석;강성원;이태훈;이해우;조수형
    • Journal of Welding and Joining
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    • v.16 no.5
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    • pp.134-141
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    • 1998
  • In this study, a fillet size for bending fatigue strength of one side fillet welded T-joint, used in box type girder and other welding structure, was investigated by bending fatigue test with or without edge preparation and burn through, with variation of joint shape. As a result, the following conclusions were obtained. (1) In one side fillet welded T-joint, the larger the leg length, the greater the bending fatigue strength. The increase in bending fatigue strength. (2) One side filet welded T-joint with edge preparation showed higher bending fatigue strength than that with twofold-large leg length and without edge preparation. (3) In one side fillet welded T-joint without edge preparation, both manual welding and automatic welding were carried out with same condition. In this case, automatic welding shoed deeper penetration and more increased horizontal leg length than manual welding, so that automatic welding offers grater bending fatigue strength. (4) For one side fillet welded T-joint without edge preparation, the ratio(h/t) of the leg length (h) and the main plate thickness (t) in which toe crack can occur was 1.2 over. (5) In one side fillet welded T-joint with edge preparation, the burn through led to reduced bending fatigue strength. However, this bending fatigue strength was higher than that of one side fillet welded T-joint without edge preparation and with a larger leg length.

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Effects of Functional Ingredients Addition on Quality Characteristics of Kochujang (기능성 소재의 첨가가 고추장의 품질특성에 미치는 영향)

  • Choi, Shin-Yang;Cho, Gyeong-Hyeon;Lim, Seong-Il
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.779-784
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    • 2006
  • To produce bio-active Kochujang, 1.2% of pomegranate powder, 1.0% of hawthorn berry extract powder, 0.01% of ginko biloba extract, 1.2% of red yeast rice powder, 1.2% of silk powder, 0.2% of garlic oil, 0.2% of chitosan, 1.2% of wax gourd extract powder, and 1.0% of pine mushroom were added individually with raw materials that are needed for Kochujang process. These Kochujangs were fermented for 45 days at $30^{\circ}C$, and examined for changes in their physicochemical properties. There were no significant differences in pH, acidity, formol-N content, color and total cell count by plate count agar between Kochujangs fortified individually with functional ingredients and the control Kochujang. When 0.2% of garlic oil was added, antioxidant activity of its Kochujang product increased twice as much as that of the control Kochujang. Improved ACE inhibiting activities were observed in 1.2% of red yeast rice powder. When 1.2% of silk powder or 0.01% of ginko biloba extract were added, antithrombotic abilities were remarkably enhanced up to two times of the control Kochujang.

Production, Immobilization, and Characterization of Croceibacter atlanticus Lipase Isolated from the Antarctic Ross Sea (남극 로스해에서 분리한 Croceibacter atlanticus균 유래 리파아제의 생산, 고정화, 효소특성 연구)

  • Park, Chae Gyeong;Kim, Hyung Kwoun
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.234-243
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    • 2018
  • The Antarctic Ocean contains numerous microorganisms that produce novel biocatalysts that can have applications in various industries. We screened various psychrophilic bacterial strains isolated from the Ross Sea and found that a Croceibacter atlanticus strain (Stock No. 40-F12) showed high lipolytic activity on a tributyrin plate. We isolated the corresponding lipase gene (lipCA) by shotgun cloning and expressed the LipCA enzyme in Escherichia coli cells. Homology modeling of LipCA was carried out using the Spain Arreo lake metagenome alpha/beta hydrolase as a template. According to the model, LipCA has an ${\alpha}/{\beta}$ hydrolase fold, Gly-X-Ser-X-Glymotif, and lid sequence, indicating that LipCA is a typical lipase enzyme. Active LipCA enzyme was purified fromthe cell-free extract by ammonium sulfate precipitation and gel filtration chromatography. We determined its enzymatic properties including optimum temperature and pH, stability, substrate specificity, and organic solvent stability. LipCA was immobilized by the cross-linked enzyme aggregate (CLEA) method and its enzymatic properties were compared to those of free LipCA. After cross-linking, temperature, pH, and organic solvent stability increased considerably, whereas substrate specificities did not changed. The LipCA CLEA was recovered by centrifugation and showed approximately 40% activity after 4th recovery. This is the first report of the expression, characterization, and immobilization of a C. atlanticus lipase, and this lipase could have potential industrial application.

Degradation of Chlorinated Phenolic Compounds by Soil Actinomycetes Isolated from the Contami-nated Soil Nearby the Kyung-An River (경안천 유역 오염토양에서 분리한 방선균의 염화 페놀계 화합물 분해)

  • 김성민;김창영;김응수
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.287-292
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    • 2002
  • Lignin-peroxidase (LiP) has been considered as one of the most important industrial enzymes for biodegradation of various recalcitrant toxic compounds such as chlorinated aromatic hydrocarbons and azo-dyes. Recently, several soil actinomycetes have been reported to secrete a functionally-similar lignin-peroxidase called actinomycetes lig-nin-peroxidase (ALiP). In this manuscript, we isolated over 100 morphologically distinct actinomycetes from the contaminated soils around 10 different gas stations located nearby the Kyung-An river. Among these actinomycetes screened based on the congo-red dye-decolorization activities, one newly-isolated actinomycetes named SMA-2 showed the most significant dye-decoloring activity on the congo-red plate as well as a significant ALiP activity in a yeast-extract-malt-extract liquid media supplemented with starch. The optimum SMA-2 culture condition fur ALiP production was determined and the kinetic parameters fur the SMA-2 AkIP activity were characterized. The optimally-cultured SMA-2 also exhibited the oxidation activities toward various recalcitrant aromatic compounds including phenol, 2- chlorophenol, 4- chlorophenol, 2,4- dichlorophenol ,2,6- dichlorophenol, and 2,4, f-trichlorophe - not, suggesting a potential application of SMA-2 for contaminated soil bioremediation.

Effects of Size and Rate of Maturing on Carcass Composition of Pasture- or Feedlot- Developed Steers

  • Brown, A.H. Jr.;Camfield, P.K.;Baublits, R.T.;Pohlman, F.W.;Johnson, Z.B.;Brown, C.J.;Tabler, G.T.;Sandelin, B.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.661-671
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    • 2006
  • Steers (n = 335) of known genetic backgrounds from four fundamentally different growth types were subjected to two production systems to study the main effects and possible interactive effects on carcass composition. Growth types were animals with genetic potential for large mature weight (LL), intermediate mature weight-late maturing (IL), intermediate mature weight-early maturing (IE), and small mature weight-early maturing (SE). Each year, in a nine year study, calves of each growth type were weaned and five steers of each growth type were developed on pasture or feedlot and harvested at approximately 20 and 14 mo of age, respectively. Data recorded were chilled carcass weight and percentages of forequarter, foreshank, chuck, rib, plate, brisket, hindquarter, round, rump, shortloin, sirloin, flank, lean, fat, bone, and retail cuts. The growth $type{\times}production$ system interaction was an important source of variation in chilled carcass weight (p = 0.0395) and percentage retail cuts (p = 0.001), lean (p = 0.001), fat (p = 0.001), rump (p = 0.0454), shortloin (p = 0.0487), and flank (p = 0.001). The ranking of the growth $type{\times}production$ system means for percentage lean was LL-pasture>IL-pasture = IE-pasture = SE-pasture>LL-feedlot, IL-feedlot>IE-feedlot = SE-feedlot. The growth $type{\times}production$ system interaction was non-significant (p>0.05) for forequarter, foreshank, chuck, rib, plate, brisket, hindquarter, round and bone. Growth types of IE and SE yielded greater (p<0.05) mean forequarter than did growth types of IL and LL ($51.6{\pm}0.3$ and $51.5{\pm}0.3$ vs. $51.1{\pm}0.3$ and $50.8{\pm}0.3%$). Mean bone was highest (p<0.05) for the LL growth type and lowest (p<0.05) for the SE growth type ($19.5{\pm}0.5$ vs. $16.8{\pm}0.5%$). Mean bone was greater (p<0.05) for the pastured steers than for the feedlot steers ($21.8{\pm}0.8$ vs. $14.5{\pm}0.6%$). These data indicate that growth type responded differently in the two production systems and that these results should be helpful in the match of genetics to production resources.

Development of a Korean Type Totally Implantable TAH (한국형 완전이식 인공심장의 개발)

  • Min, B.G.;Choi, W.W.;Ahn, J.M.;Park, S.K.;Park, C.Y.;Chang, J.K.;Kim, J.W.;Kim, H.C.;Kim, W.K.;Roh, R.J.
    • Proceedings of the KOSOMBE Conference
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    • v.1996 no.05
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    • pp.199-202
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    • 1996
  • Artificial hearts are intended for use in patients with severe forms of heart disease for which no surgical repair is possible. The moving-actuator pump was developed to decrease the overall volume size of the electromechanical total artificial heart (TAH) by eliminating the occupied space of the fixed-actuator in the conventional pusher-plate type pump. In our pump, the actuator moves back and forth for alternative ejections of left and right ventricles. The problem of fitting the TAH to atrial remnants and arterial vessels could also be improved by circular or penduluous mot ion of the actuator instead of linear mot ion of the pusher-plate in the conventional pumps. We have evaluated two types of moving- actuator pump; one is a rolling cylinder type, and the other a pendulum type pump. In the rolling cylinder pump, frictional energy loss exists between the pump housing's guide bars and the actuator's end caps, while the bottom rack under the cylindrical actuator increases the height of the pump, the pump is therefor not implantable inside the small chest of human-sized animals with a body weight of less than 70kg. The new human type pump has a penduluous mot ion actuator to correct the above problems while maintaining the advantage of the moving- actuator's small total volume. The totally implantable TAH is composed of a blood pump, a control system and pheriperal equipments. The blood pump, which is constructed by a moving actuator, a right and left blood sac, and four artificial valves, is implanted in the thoracic. In 1988, the first implantation of the rolling cylinder TAH was performed into a female calf weighing 100kg, and the cal f recovered to the degree of voluntary standing and eat ing and survived to 100 hrs. We then survived two female sheep weighing about 63kg with the new human type TAH for three days.

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Isolation of Antifungal Substances by Bacillus amyloliquefaciens IUB158-03 and Antagonistic Activity against Pathogenic Fungi (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 분리와 항균활성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.96-103
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    • 2009
  • For the control of pathogenic microorganisms, Bacillus spp. were isolated from diseased pepper fruits in Korea. Among them, Bacillus sp. IUB158-03 showed high inhibitory effect on mycelial growth and spore germination of C. gloeosporioides and Botrytis cinerea. The strain was identified as B. amyloliquefaciens IUB158-03 based on its physiological, biochemical characteristics and Microlog analysis. The highest level of antifungal substances by B. amyloliquefaciens IUB158-03 were obtained when the bacterium was cultured in medium containing 2% soluble starch, 3% yeast extract, 0.5% tryptone, 0.5% $NH_4H_2PO_4$, and 1% NaCl (pH 6.0) at $25^{\circ}C$ for 72 hrs. The antifungal substances were purified by butanol extraction, silica gel column chromatography, preparative thin layer chromatography, and high performance liquid chromatography. The purified antifungal substance was confirmed $R_f$ 0.27 by TLC. This substance exhibited antifungal activity against Fusarium solani, Rhizoctonia solani, Botrytis cineria, Alternata alternaria of plant pathogenic fungi and Trichophyton mentagrophytes, Epidermophyton floccosum, Cryptococcus neoformans of human pathogenic fungi.

Effects of Embryo Sources and Culture Systems on the Membrane Permeability and Viability of Bovine Blastocysts Cryopreserved by GMP Vitrification (소 수정란의 생산체계가 세포막 투과력 및 GMP Vitrification 동결융해 후 생존성에 미치는 영향)

  • Kong, I.K.;Cho, S.G.
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.191-198
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    • 2001
  • The purpose of this study was to investigate the effects of embryo sources such as in vivo vs. in vitro produced blastocyst, and culture systems on the membrane permeability and viability of bovine blastocyst following GMP vitrification. To produce in vivo embryos, six cows were superovulated by administration of follicle stimulation hormone (FSH) and prostaglandin $F_{2{\alpha}}$(PG $F_{2{\alpha}}$). in vitro embryos were produced by two different culture systems, oviduct co-culture (OCS) and defined culture system (HECM-6; DCS). Ovaries were picked up at a local slaughterhouse and transported to laboratory in 3$0^{\circ}C$ saline within 2 h. Ovaries were washed with same saline three times and then placed in saline on warm plate adjusted at 3$0^{\circ}C$ during aspiration. The blastocysts produced were assigned for membrane permeability and viability following GMP vitrification. The membrane permeability of blastocysts was checked in 0.5 M sucrose solution on warm plate at 35$^{\circ}C$ for 0, 2, 5 and 7 min, respectively. Then the diameters (width and length) of embryo cytoplasms were measured by a eyepiece meter, and they were converted to their volume by 4/3 $\pi$ $r^3$. The blastocysts were cryopreserved by GMP vitrification method, where they were sequentially placed into vitrification solution before being loaded into GMP vessels and immersed into L$N_2$ within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for 5 min each, and then cultured in TCM 199 supplemented with 10% FCS for 24 or 48 h. The volume change of in vivo blastocyst at 0, 2, 5 and 7 min (100, 37.1, 34.3 and 31.6%) was significantly more shrunk than those of in vitro blastocysts derived from OCS (100, 59.8, 48.9, 47.9%) and DCS (100, 57.2, 47.3 and 46.9%) (P<0.05). The viability of post-thaw blastocyst derived from in vivo (93.6%) was also significantly different from those in OCS and DCS (81.9 and 83.6%; P<0.05). In the present culture system, the morphology of embryos produced in vitro was similar to that of in vivo embryos, but the quality in membrane permeability and post-thaw viability showed a big difference from their sources as in vivo or in vitro derived from OCS and DCS. The results indicated that the quality of in vivo embryos in membrane permeability and post-thaw viability was better than those of in vitro embryos derived from OCS or DCS.

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Interfacial Electrical Studios on Adhesion of Hematite Particles to Polyester Fabric and their Removal from the Fabric(Part 2) (Polyester 직물에의 Hematite 입자의 부착과 제거에 관한 계면 전기적고찰 (제2보))

  • 강인숙;김성련
    • Journal of the Korean Society of Clothing and Textiles
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    • v.19 no.5
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    • pp.765-773
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    • 1995
  • Effect of interfacial electrical conditions such as, the f potential of PET fiber and u-Fe203 particles, the stability parameter and potential energy of interaction on adhesion of a-Fe903 particles to PET fabric and their removal from the fabric, were investigated as functions of pH, electrolyte and ionic strength. The stability parameter, potential energy of interaction between a-Fe2O3 particles and PET fabric were calculated by using the heterocoagulation theory for a sphere-plate model The adhesion of a-Fe2O3 particles to PET fabric and their removal from PET fabric were carried out by using water bath shaker and Terg-O-Tometer under various solution conditions. The adhesion of a-Fe2O3 particles to the PET fabric and the removal of a-Fe2O3 particles from the PET fabric were biphasic and were maximum and minimum at pH 7~8, respectively. With high pH and polyanion electrolytes in solution, the adhesion of a-Fe2O3 particles to the PET fabric was low but effects of electrolytes on the removal of a-Fe2O3 particles from the PET fabric was small. The adhesion of a-Fe2O3 particles to the PET fabric and the removal of a-Fe2O3 Particles from the PET fabric were biphasic, and were lowest and highest at the ionic strength 1$\times$10-3, respectively. The adhesion of a-Fe2O3 particles to PET fabric was well related with the interfacial electrical conditions; it was negatively correlated with the f potentials of a-Fe2O3 Particles of its absolute value, the stability parameter and the maximum of total potential energy, while, the adhesion was not related with the t potentials of PET fiber itself. Therefore, the primary factor determining the adhesion of a-Fe203 particles to PET fabric may be the stability of dispersed particles caused by the electrical repulsion of particles. The removal of a-Fe203 particles from PET fabric was not related to such interfacial electrical conditions as the t potentials of PET fiber, the stability parameter and the maximum of total potential energy but removal was related to t potential of a-Fe203 particles.

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