• Journal of Animal Science and Technology
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• 제62권2호
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• pp.141-158
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• 2020

Skin is a major thermoregulatory organ in the body controlling homeothermy, a critical function for climate adaptation. We compared genes expressed between tropical- and temperate-adapted cattle to better understand genes involved in climate adaptation and hence thermoregulation. We profiled the skin of representative tropical and temperate cattle using RNA-seq. A total of 214,754,759 reads were generated and assembled into 72,993,478 reads and were mapped to unique regions in the bovine genome. Gene coverage of unique regions of the reference genome showed that of 24,616 genes, only 13,130 genes (53.34%) displayed more than one count per million reads for at least two libraries and were considered suitable for downstream analyses. Our results revealed that of 255 genes expressed differentially, 98 genes were upregulated in tropically-adapted White Fulani (WF; Bos indicus) and 157 genes were down regulated in WF compared to Angus, AG (Bos taurus). Fifteen pathways were identified from the differential gene sets through gene ontology and pathway analyses. These include the significantly enriched melanin metabolic process, proteinaceous extracellular matrix, inflammatory response, defense response, calcium ion binding and response to wounding. Quantitative PCR was used to validate six representative genes which are associated with skin thermoregulation and epithelia dysfunction (mean correlation 0.92; p < 0.001). Our results contribute to identifying genes and understanding molecular mechanisms of skin thermoregulation that may influence strategic genomic selection in cattle to withstand climate adaptation, microbial invasion and mechanical damage.

• 대한화학회지
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• 제24권1호
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• pp.25-33
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• 1980

토양에서 분리한 Pseudomonadaceae 속 박테리아로부터 pyrocatechase를 추출, 분리 정제하였으며, 이 효소의 특성을 검토한 결과 pyrocatechase는 catechol에 대하여 기질 특이성을 보여줌을 알았다. 효소 활성도의 최적조건은 pH 7∼10 부근과 온도 $35^{\circ}C$임을 알았으며, catechol에 대한 $K_m$값은 $1.9{\times}10^{-6}M$ 로 얻어졌다. 기질 유도체에 의한 효소 저해 실험결과 벤젠 고리의 ortho 위치에 두개의 수산기는 효소-기질간의 결합반응에 참여될 것이라고 추측했다. 기타 SH-잔기와 작용하는 화합물 또는 중금속 이온등의 첨가에 따른 효소 활성도의 저해 효과를 검토 하였으며, 효소 활성부위에 대하여도 검토해 보았다.

• Journal of Microbiology
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• 제45권4호
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• pp.333-338
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• 2007

Intracellular NADH:quinone reductase involved in degradation of aromatic compounds including lignin was purified and characterized from white rot fungus Trametes versicolor. The activity of quinone reductase was maximal after 3 days of incubation in fungal culture, and the enzyme was purified to homogeneity using ion-exchange, hydrophobic interaction, and gel filtration chromatographies. The purified enzyme has a molecular mass of 41kDa as determined by SDS-PAGE, and exhibits a broad temperature optimum between $20-40^{\circ}C$, with a pH optimum of 6.0. The enzyme preferred FAD as a cofactor and NADH rather than NADPH as an electron donor. Among quinone compounds tested as substrate, menadione showed the highest enzyme activity followed by 1,4-benzoquinone. The enzyme activity was inhibited by $CuSO_4,\;HgCl_2,\;MgSO_4,\;MnSO_4,\;AgNO_3$, dicumarol, KCN, $NaN_3$, and EDTA. Its $K_m\;and\;V_{max}$ with NADH as an electron donor were $23{\mu}M\;and\;101mM/mg$ per min, respectively, and showed a high substrate affinity. Purified quinone reductase could reduce 1,4-benzoquinone to hydroquinone, and induction of this enzyme was higher by 1,4-benzoquinone than those of other quinone compounds.

• 한국재료학회지
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• 제16권11호
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• pp.692-697
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• 2006

Deodorization of natural zeolites have been improved not only for polar but also for non-polar pollutants by sucessive ion exchanges of H and Ag ions starting from Korean natural zeolite with high adsorption capacity. The modified zeolites with $TiO_2$ coating on the surface revealed high deodorization and photocatalytic decomposition effects. Further modification was made with $10{\sim}20nm$ silica nano particles coating on the surface, the resulting composite particles of $SiO_2/TiO_2/modified$ natural zeolite revealed not only comparable deodorization but also better durability and resisatnce to color change compared to the $TiO_2$/modified natural zeolite without much compensation of photocatalytic decomposition effect, when the composite particles were exposed to the polypropylene non-woven fiber coated with organic binder. It is expected for the composite particle prepared here to be used as indoor building materials for indoor air quality control.

• Food Science and Biotechnology
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• 제17권4호
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• pp.766-771
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• 2008

The proteolytic enzyme from Saccharomyces sp. B101 was purified to homogeneity by ammonium sulfate fractionation, ultrafiltration, diethyl aminoethyl (DEAE)-Sephadex A-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography from the culture supernatant of Saccharomyces sp. B101. The specific activity and the purification fold of the purified enzyme were 4,688.9 unit/mg and 18, respectively. The molecular weight of the purified enzyme was estimated to be 33 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature for the enzyme activity were pH 8.5 and $30^{\circ}C$, respectively. The enzyme activity was relatively stable in the pH range of 6.5-8.5 at below $35^{\circ}C$. The salt-tolerance and stability for the enzyme activity were relatively stable even at NaCl concentrations of 10 and 15%. The activity of enzyme was inhibited by $Ag^{2+}$ and $Fe^{2+}$, and activated by $Mn^{2+}$. In addition, the enzyme activity was potently inhibited by ethylenediaminetetraacetic acid (EDTA) and phenylmethyl sulfonylfluoride (PMSF). Based on these findings we concluded that the purified enzyme was a serine protease. Km and Vmax values for hammastein milk casein were 1.02 mg/mL and 278.38 unit/mL, respectively.

• 산업기술연구
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• 제39권1호
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• pp.15-19
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• 2019

This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.

• 생명과학회지
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• 제18권6호
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• pp.789-795
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• 2008

어류 질병 치료를 위한 probiont의 개발에 목적을 두고 어병균 Vibrio anguillarum NCMB1의 생육에 저해물질을 생산하는 Bacillus amyloliquefaciens H41균주를 분리하고 이 물질의 특성을 규명하기 위하여 정제를 시행하였다. 분리 균을 배양한 배양 상등액을 70% 염석, 투석하여 조 효소액으로 제작하고 조 효소액을 DEAE-sephadex, A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography을 통하여 정제하고 SDS-PAGE 를 통하여 단일밴드를 확인하고 최종 회수율 2.9%을 얻을 수 있었으며 40.8배의 정제된 V. anguillarum NCMB1 생육저해물질을 얻을 수 있었다. 정제된 저해물질은 저해정도에 따라 단위를 설정하여 활성을 측정하였으며, 분자량은 48 kDa 로 확인되었으며 정제물질의 활성을 위한 최적 반응 pH와 온도는 pH 7.5와 $30^{\circ}C$로 확인되었다. 금속이온의 효과에 있어서는 $CoCl_2$, $HgCl_2$, $ZnSO_4$, $AgNo_3$에서는 완전히 저해되는 양상을 나타내었고 $MgSO_4$, $MnSO_4$에서 미미한 효소활성의 증가를 나타내었다. 그리고 염에 관한 안정성은 일반 해수의 농도인 3%의 농도에서도 활성을 나타내는 것으로 확인되었다. 정제된 저해물질을 현재 상업적으로 사용하고 있는 화학처리제나 항생제와 함께 효율성을 비교 해 보았을 때 저해물질은 약 78%의 저해 활성을 나타내는 것을 알 수 있었고 항생제보다는 효율성이 낮았으나 독성검사를 위해 정제물질을 살아있는 어류에 투여하였으나 어떤 해수어도 폐사하지 않는 것으로 보아 어류 자체엔 독성을 나타내지 않는 물질로 나타났다. 따라서 B. amyloliquefaciens H41 균주가 생산하는 정제 물질이 V. anguillarum 생육에 저해물질로 작용하는 것으로 밝혀졌으며 친환경적인 특성을 가진 물질로 밝혀졌다.

• 한국응용과학기술학회지
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• 제18권3호
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• pp.197-213
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• 2001

Conjugate trienoic acids (CTA) occurred in triacylglycerols (TGs) of the seed oils of Trichosanthes kirilowii, Momordica charantia and Aleurites fordii, and they were easily converted to their methyl esters in a mixture of sodium methoxide-methanol without any structural destruction. The main fatty acids in triacylglycerol (TG) fraction of the seed oils of Trichosanthes kirilowii are $C_{18:2{\omega}6}$ (32.2 mol %), $C_{18:3{\;}9c.11t,13c}$ (38.0 mol %) and $C_{18:1{\omega}9}$ (11.8 mol %), followed with $C_{16:0}$ (4.8 mol %) and $C_{18:0}$ (3.1 mol %). The TG fraction was resolved into 20 TG molecular species according to the partition number (PN) by reversed-phase (RP)-HPLC. The main TG species were $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$, of which amounts reached 63 mol % of total TG molecular species. The TG sample was fractionated into 11 fractions according to the number of double bond in the molecule by $Ag^{+}-HPLC$ and the species of $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$ were also eluted as main components. The TG species containing CTA showed unusual behaviours in the order of elution by HPLC ; first, TG moleular species of $DT_{c2}$ (D; dienoic acid, $T_{c}$; punicic acid, $T_{ci}$; ${\alpha}-eleostearic$ acid, M ; monoenoic acid, $S_{t}$; stearic acid) was eluted earlier than $Mt_{c2}$, although they have the same PN number of 40, and, secondly, the species of $DT_{ci2}$ with eight double bonds was eluted earlier than that of $D_2T_{ci}$ with seven double bonds. Intact TG of the seed oils of Momordica charantia contained mainly fatty acids such as $C_{18:3{\omega}9c,11t,13t}$ (57.7 mol %), $C_{18:1{\omega}9}$ (17.4 mol %), $C_{18:0}$ (12.3 mol %) and $C_{18:2{\omega}6}$ (10.6 mol %), and was classified into 13 fractions by RP-HPLC. The main TG species were as follows ; $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.1 mol %] and $S_{t}T_{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_2$, 33.9 mol %] comprising about 73 mol % of total TG species, accompanied by $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 7.3 mol %], $D_{2}T_{ci}$ [$ (C_{18:2{\omega}6})_{2}(C_{18:3\;9c,11t,13t})$, 3.6 mol %] and $MDT_{ci}$ [$(C_{18:1{\omega}9})(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})$, 3.5 mol %]. Simple TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_3]$ was present in a small amount of 1.4 mol %, but other simple TG species were not detected. The TG was also resolved into 11 fractions according to the number of double bond by $Ag^{+}-HPLC$, and the species were mainly occupied by $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.4 mol %] and $S_tT-{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_{2}$, 35.4 mol %] $DT_{ci2}$ species with eight double bonds was also developed faster than $D_2T_{ci}$ one with seven double bonds as indicated in the analysis of TG of the seed oils of T. kirilowii, and $MT_{ci2}$ species with cis, trans, trans-configurated double bond was eluted earlier than $MT_{c2}$ species with cis, trans, cis-configurated double bond. The main components of fatty acid in total TG fraction isolated from the seed oils of of Aleurites fordii were in the following order ; $C_{18:3\;9c,11t,13t}$ (81.2 mol %)> $C_{18:2{\omega}6}$ (8.5 mol %)> $C_{18:1{\omega}9}$ (5.4 mol %)$. With resolution of the TG by RP-HPLC, eight fractions such as $T_{ci3}$, $Dt_{ci2}$, $D_{2}T_{ci}$, $MT_{ci2}$, $PT_{ci2}$ (P; palmitic acid), $PMT_{ci}$, $PDT_{ci}$ and $S_{t}T_{ci2}$ ($S_{t}$; stearic acid) were isolated, respectively. TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_{3}$, 54.2 mol %], $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 15.0 mol %] and $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3 9c,11t,13t})_{2}$, 14.8 mol %] were present as main species.

• Restorative Dentistry and Endodontics
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• 제22권1호
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• pp.1-33
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• 1997

The objective of this study was to analyze the in vitro and in vivo corrosion products of low and high copper amalgams. The four different types of amalgam alloy used in this study were Fine cut, Caulk spherical, Dispersalloy, and Tytin. After each amalgam alloy and Hg were triturated according to the directions of the manufacturer by means of the mechanical amalgamator(Amalgam mixer. Shinhung Co. Korea), the triturated mass was inserted into a cylindrical metal mold which was 12mm in diameter and 10mm in height. The mass was condensed by 150Kg/cm compressive force. The specimen was removed from the mold and aged at room temperature for about seven days. The standard surface preparation was routinely carried out by emery paper polishing under running water. In vitro amalgam specimens were potentiostatically polarized ten times in a normal saline solution at $37^{\circ}C$(potentiostat : HA-301. Hukuto Denko Corp. Japan). Each specimen was subjected to anodic polarization scan within the potential range -1700mV to+400mV(SCE). After corrosion tests, anodic polarization curves and corrosion potentials were obtained. The amount of component elements dissolved from amalgams into solution was measured three times by ICP AES(Inductive Coupled Plasma Atomic Emission Spectrometry: Plasma 40. Perkim Elmer Co. U.S.A.). The four different types of amalgam were filled in occlusal and buccal class I cavities of four human 3rd molars. After about five years the restorations were carefully removed after tooth extraction to preserve the structural details including the deteriorated margins. The occlusal surface, amalgam-tooth interface and the fractured surface of in vivo amalgam corrosion products were analyzed. In vivo and in vitro amalgam specimens were examined and analyzed metallographically by SEM(Scanning Electron Microscope: JSM 840. Jeol Co. Japan) and EDAX(Energy Dispersive Micro X-ray Analyser: JSM 840. Jeol Co. Japan). 1. The following results are obtained from in vitro corrosion tests. 1) Corrosion potentials of all amalgams became more noble after ten times passing through the in vitro corrosion test compared to first time. 2) After times through the test, released Cu concentration in saline solution was almost equal but highest in Fine cut. Ag and Hg ion concentration was highest in Caulk spherical and Sn was highest in Dispersalloy. 3) Analyses of surface corrosion products in vitro reveal the following results. a)The corroded surface of Caulk spherical has Na-Sn-Cl containing clusters of $5{\mu}m$ needle-like crystals and oval shapes of Sn-Cl phase, polyhedral Sn oxide phase. b)In Fine cut, there appeared to be a large Sn containing phase, surrounded by many Cu-Sn phases of $1{\mu}m$ granular shapes. c)Dispersalloy was covered by a thick reticular layer which contained Zn-Cl phase. d)In Tytin, a very thin, corroded layer had formed with irregularly growing Sn-Cl phases that looked like a stack of plates. 2. The following results are obtained by an analysis of in vivo amalgam corrosion products. 1) Occlusal surfaces of all amalgams were covered by thick amorphous layers containing Ca-P elements which were abraded by occlusal force. 2) In tooth-amalgam interface, Ca-P containing products were examined in all amalgams but were most clearly seen in low copper amalgams. 3) Sn oxide appeared as a polyhedral shape in internal space in Caulk spherical and Fine cut. 4) Apical pyramidal shaped Sn oxide and curved plate-like Sn-Cl phases resulted in Dispersalloy. 5) In Tytin, Sn oxide and Sn hydroxide were not seen but polyhedral Ag-Hg phase crystal appeared in internal space which assumed a ${\beta}_l$ phase.

• 한국수산과학회지
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• 제19권6호
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• pp.547-557
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• 1986

전보(Pyeun and Kim, 1986)에서와 같이 고등어 유문수조직에서 정제한 3종의 알칼리성단백질분해효소에 대하여 활성최적조건, 열안정성. 기질친화도, 화학약제에 대한 영향 및 분자량 등을 규명하였다. 각 정제효소의 반응최적조건을 기질별로 검토한 결과, casein에 대하여 Enz. A는 pH 9.4, Enz. B와 Enz. C는 pH 9.8이었으며, Hb에 대하여 Enz. A는 pH 9.2, Enz. B는 pH 10.2, 그리고 Enz. C는 pH 9.8이었고, 최적반응온도는 공히 $45^{\circ}C$였다. 효소농도 $2{\mu}g/ml,\;2\%$ casein 기질의 반응조건에서 반응시간(x)에 대한 활성도 (y)의 관계를 분석한 결과, Enz. A는 60분. Enz. B는 40분, 그리고 Enz. C는 50분까지 1차 반응의 관계가 성립하였으며, 이때의 반응속도식은 Enz. A는 y=3.6x, Enz. B는 Y=6.0x, 그리고 Enz. C는 y=4.2x였다. 열안정성을 검토하기 위하여 $50^{\circ}C$에서 5분간 가열했을때, Enz, A는 $90\%$, Enz. B는 $33\%$, 그리고 Enz. C는 $37\%$가 각각 불활성화하였다. Lineweaver-Burk의 도식에 의한 효소의 기질친화도를 측정한 결과, casein기질에 대하여 Enz. A는 Km이 $5.0{\times}10^{-3}\%$, Enz. B는 Km이 $1.0{\times}10^{-3}\%$, 그리고 Enz. C는 Km이 $3.6{\times}10^{-3}\%$였다. 금속 ion에 의한 영향을 검토한 결과, $Ag^+,\;Hg^{2+}$는 효소활성을 저하시켰으나, $Mn^{2+},\;Sn^{2+}$ 및 $Pb^{2+}$ 이온은 활성을 증가시켰다. Enz. B와 Enz. C는 soybean trypsin inhibitor에 의해 상당히 저해되었다. 따라서 효소 B와 C는 serine 계 단백질분해효소로 판단되었다. SDS-PAG 전기영동과 Sephadex G-100 겔 여과법에 의하여 각 정제효소의 분자량을 측정한 결과, Enz. A는 $27,500{\pm}2.500$, Enz. B는 $20,500{\pm}1,500$, 그리고 Enz. C는 $15,250{\pm}250$이었다.