• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.230-237
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• 2004

In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{＋}$ , $Cu^{ 2+}$, $Hg^{2＋}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2＋}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• Economic and Environmental Geology
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• v.32 no.5
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• pp.445-454
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• 1999

The Sanjeon Au-Ag deposit consists of three subparallel hydrothermal quartz-calcite veins which filled fault-related fractures (generally $N20^{\circ}$ to 35"W-trending and $70^{\circ}$ to $80^{\circ}$ SW-dipping) within quartz porphyry. The vein mineralization shows an apparent variation of mineral assemblages with paragenetic time: (1) early, white quartz + pyrite + arsenopyrite + brown sphalerite, (2) middle, white (vein) to clear quartz (vug) + base-metal sulfides + electrum + argentite, (3) late, calcite + pyrite + native silver. Mineralogic and fluid inclusion data indicate that gold-silver minerals were deposited at temperatures from 2l $0^{\circ}$ to $250^{\circ}$ with salinities of 4 to 5 wt. % equiv. NaCl and log fS2 values from -14.0 to -12.2 atm. The linear relationship between homogenization temperature and salinity data indicates that gold-silver deposition was a result of meteoric water mixing. Ore mineralization occurred at pressure conditions of about 70 bars, which corresponds to the mineralization depths of about 260 m to 700 m. There is a remarkable decrease of the calculated 1)180 values of water from 1.3 to -9.7%0 in hydrothermal fluid with increasing paragenetic time. This indicates a progressive increase of meteoric water influx in the hydrothermal system at the Sanjeon deposit. Oxygen-hydrogen, sulfur, and carbon isotope values of hydrothermal fluids indicate that the ore mineralization was formed largely from meteoric waters with the contribution of sulfur and carbon from a deep igneous source.

• Proceedings of the Korean Institute of Surface Engineering Conference
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• 2018.06a
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• pp.106-106
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• 2018

해양환경용 선박재료는 전기화학적인 부식을 발생시키는 염소이온($Cl^-$)이 다량 포함된 부식 환경에 장기간 노출되어 있어 부식에 대해 취약하다. 따라서 우수한 내식성 및 내침식성을 가진 재료를 선정하는 것은 매우 중요하다. 알루미늄 합금은 충분한 강도와 부동태 피막 형성으로 인해 내식성이 우수하여 해양환경용 선박 재료로서 널리 이용되고 있으며, 이에 따른 부식 특성에 관한 연구도 활발히 이뤄지고 있다. 그러나 선박에서는 부식에 의한 손상뿐만 아니라 전식에 의한 부식 손상도 발생할 수 있다. 특히 선미 부분은 프로펠러의 동합금과 알루미늄 합금의 이종금속 간 전위차에 의한 전식이 발생하여 선체의 다른 부위에 비해 부식이 더 심하게 진행될 수도 있다. 또한 전식은 해안 부두에 접안된 선박의 용접 시미주전류(stray current)에 의한 부식손상이 발생할 수 있으나 이에 대한 연구는 미미한 실정이다. 따라서 본 연구는 해양환경에서의 전식을 인위적으로 모사할 수 있는 부식 정전류 시험법을 이용하여 다양한 크기의 전식 손상을 유발시켰으며, 해양환경 하에서 선박재료로 주로 사용되는 알루미늄 합금인 Al5083-H321, Al5052-O, Al6061-T6에 대한 전식 특성을 비교, 분석하였다. 실험 방법으로 작동전극은 각 재료의 시험편을 $2cm{\times}2cm$ 으로 절단하여 sand paper # 2000 번까지 연마 후 아세톤과 증류수로 세척하고 건조하였으며, 제작된 시험편은 자체 제작한 홀더를 이용하여 $1cm^2$만 노출시킨 후 정전류 가속 실험을 실시하였다. 기준전극은 은/염화은(Ag/AgCl) 전극을, 대응전극은 백금(Pt) 전극을 사용하였다. 정전류 가속 조건은 $0.001mA/cm^2$, $0.1mA/cm^2$, $1mA/cm^2$, $5mA/cm^2$, $10mA/cm^2$의 전류 밀도를 천연해수에서 30분간 인가하였다. 각 재료에 대한 전식 특성은 실험 전후의 무게 감소량으로 전식의 저항 특성을 확인하였다. 그리고 3D 현미경으로 표면 손상 경향과 깊이를 측정하였으며, 주사전자현미경 (SEM)을 통해 표면 형상을 미시적으로 관찰하였다. 부식 정전류 시험 결과 모든 시편에서 $0.01mA/cm^2$에서 미세한 국부적인 부식이 일어났으며, 전류밀도가 증가할수록 표면 전반에 부식이 진행되고 성장하였다. 그리고 모든 인가 전류밀도의 조건에서 Al6061-T6가 5000계열(Al5083-H321, Al5052-O)보다 더 우수한 내식성을 나타났다.

• Applied Biological Chemistry
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• v.34 no.1
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• pp.54-60
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• 1991

Serratia marcescens CK-3, decomposing chitin which is a mar component of cell wall in phyitopathogenic fungi, was isolated from the continuous cropping rhizosphere of pepper and cucumber and its enzymatic property was examined. S. marcescens CK-3 was found tn have an tagonistic effects against, Fusarium axysporum and Rhizoctonia solani and to have complex enzyme system such as chitinase, laminarinase, and proteinase. The preferable composition of the medium for production of chitinase was fond and was as follows : colloidal chitin 1.5%, tryptone 0.5%, glucose 1.0%, peptone 0.2%, $MgSO_4{\cdot}7H_2O\;0.1%,\;K_2HPO_4\;0.1%,\;and\;NaCl\;0.1%$(w/v), pH 6.8. The maximum enzyme production was observed after culture of 72 hours at $30^{\circ}C$ using a medium containing the above chemical composition. The optimal pH and temperature for in vitro activity of chitinase from S. marcescens CK-3 were pH 7.5 and $50^{\circ}C$, respectively. The enzyme activity in-creased by metal ions such as$Ag^+$ and $Mn^{++}$.

• Economic and Environmental Geology
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• v.25 no.3
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• pp.233-243
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• 1992

Ore mineralization of the Hwanghak copper deposit in the Ogsan area occurred in three stages of quartz (stage I and II) and calcite (stage III) veining along fissures in Early Cretaceous sedimentary rocks. Ore minerals are pyrite, pyrrhotite, chalcopyrite (dominant), sphalerite, hematite, galena, and Ag-, Pb-, and Bi-sulfosalts. These were deposited during the first stage at temperatures between $370^{\circ}C$ and < $200^{\circ}C$ from fluids with salinities between 0.5 and 7.6 equiv. wt. % NaCl. There is evidence of boiling and this suggests pressures of less than 180 bars during the first stage. Equilibrium thermodynamic interpretation accompanying with mineral paragenesis and fluid inclusion data indicates that copper precipitation in the hydrothermal system occurred due to cooling and changing in chemical conditions ($fs_2$, $fo_2$, pH). Gradual temperature decrease from $350^{\circ}$ to $250^{\circ}C$ of ore fluids by boiling and mixing with less-evolved meteoric waters mainly led to copper deposition through destabilization of copper chloride complexes. Sulfur isotope values of sulfide minerals decrease systematically with paragenetic time from calculated ${\delta}^{34}S_{H_2S}$ values of 8.2 to 4.7‰. These values, together with the observed change from sulfide-only to sulfide-hematite assemblages and fluid inclusion data, suggest progressively more oxidizing conditions, with a corresponding increase of the $sulfate/H_2S$ ratio of hydrothermal fluids. Measured and calculated hydrogen and oxygen isotope valutls of ore-forming fluids suggest meteoric water dominance, approaching unexchanged meteoric water values.

• Korean Journal of Food Science and Technology
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• v.22 no.1
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• pp.13-18
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• 1990

Crude lipase activator (L. Activator) was extracted with 0.85M NaCl solution from green pepper, Capsicum annuun Lin and then fractionated by 0.2 saturation with ammonium sulfate. The activity of crude L. Activator preparation $(OD_{280}=1.0)$ had proportional relation with its added amounts below 1.0ml. The L.Activator showed optimum temperature at $35^{\circ}C$. The L.Activator was very stable at the temperatures below $50^{\circ}C$ and at pH range of $7{\sim}9$, and its activities also remained 60% even at $100^{\circ}C$, 72% at pH 3, and 85% at pH 10, respectively. The activities of L.Activator decreased by most metal ions besides $Na^+,\;Mg^{++},\;and\;Ca^{++}$. The decreasing effects of heavy metal ions such as $Ag^+\;and\;Hg^{++}$ on L.Activator activity were not, however, so great as compared with the commonly known great effects of them on most enzyme activity. Crude L.Activator was separated into 4 peaks by the cellulofine column chromatography and the main active peak of L.Activator seemed to be contained in the same components as those of the activatory peak from crude L.Inhibitor.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.74-80
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• 2002

The purpose of this work was to investigate the characterization and sequence of catechol 1,2-dioxygenase (Cl,2O) purified from Acinetobacter sp. KS-1 which was grown on benzoate as a sole carbon source. Cl,2O demonstrated its enzyme activity to catechol and 4-methylcatechol. The optimum temperature of Cl,2O was $35^{\circ}C$, and the optimal pH was in the range from pH 7.5 to 9.0. $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of Cl,2O. Molecular weight of the enzyme was determined to approximately 36 kDa by SDS-PAGE and 7-terminal amino acid sequence of Cl,2O was analyzed as $^{1}MNYQQIDALVKQMNVDTAKG^{20}$and exhibited 95% sequence homology with that of Cl,2O from Acinetobacter radioresistens In addition, trypsin digestion and peptide mapping were performed for internal sequencing analysis. Molecular weights of three digested peptide fragments were analyzed as 966.3 Da, 1933.8 Da and 2081.7 Da by MALDI-TOF, which were matched with each internal sequences $^{1}SQSDFNLRR^{9}\, ^{1}HGNRPSHVHYFNSAPGYR^{18}\, ^{1}TIEGPLYVAGAPESVGFAR^{19}$) of. A. radioresistens. PCR product was amplified with the degenerated primers derived from N-terminal and each internal amino acid sequences.

• Journal of Life Science
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• v.14 no.1
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• pp.110-114
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• 2004

The optimized RAPD-PCR conditions, that can be utilized as a basic information for analysis of the gelletic characteristics were developed for genetic analysis of four mulberry varieties, named Milsung, Chungil, Suil, and Hansung using a primer, OPY15 (5'-AGTCGCCCTT-3') from Operon company. We tested several different factors for best PCR condition including concentrations of DNA, primer, Mgclu annealing temperature, number of PCR cycle, and prosence/absence of pre-heating time at the begining of PCR reaction in the $25 \mul$volume. The best RAPD profiles were obtained using 50 ng of DNA, 1 $\mu$M of primer, $1 \mum$of $MgCl_2\;,45^{\circ}C$ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the mulberry varieties, re-establishment of phylogenetic relationships and development of new varieties.

• YAKHAK HOEJI
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• v.37 no.2
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• pp.129-135
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• 1993

Serratia sp. Y-4 was isolated from soil. Many characteristics of the strain and optimum cultivation condition for protease production were investigated.,The protease from Serratia sp. Y-4 was purified and studied for the properties of the enzyme. The isolated strain was identified to the genus Serratia. The strain was cultivated in 1%-casein, 0.5%-Na$_{3}$PO$_{4}$.7H$_{2}$O, 0.1%-NaCl, 0.05%-KCI, 0.02%-MgSO$_{4}$.7H$_{2}$O, 0.02%-CaCl$_{2}$.2H$_{2}$O, 0.02%-ZnSO$_{4}$.7H$_{2}$O, 0.02%-MnCl$_{2}$.4H$_{2}$O and 0.5%-soy bean oil at pH 7.0 for 35 hrs. The enzyme was purified about 5.89 fold from the culture broth with 31.1% recovery and 19,613 u/mg through ultrafiltration, ammonium sulfate, DEAE-sephacel and Superose-12 chromatography. The purified enzyme was identified as one band by isoelectric focusing, SDS- and native-PAGE. It has maxium activity at $37^{\circ}C$ and pH 9.0. Molecular weight of it is approx. 50 kD and pl is about 6.70. Its Km value for casein was 20 mg/ml. 5 mM-EDTA, 5mM-SDS, Ag$^{+1}$, Cu$^{+2}$, Hg$^{+2}$ and Pb$^{+2}$ inhibited the enzyme.