• Bulletin of the Korean Chemical Society
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• 제31권11호
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• pp.3212-3216
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• 2010

A new rhodamine-based sensor 1 was designed and synthesized by incorporating rhodamine B and benzimidazole moieties. Sensor 1 exhibits high selectivity and sensitivity to $Cu^{2+}$ in $CH_3CN$-water solution (HEPES buffer, pH = 7.0) with an obvious color change from colorless to pink. Other metal ions such as $Hg^{2+}$, $Ag^+$, $Pb^{2+}$, $Sr^{2+}$, $Ba^{2+}$, $Cd^{2+}$, $Ni^{2+}$, $Co^{2+}$, $Fe^{2+}$, $Mn^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Ce^{3+}$, $Mg^{2+}$, $K^+$ and $Na^+$ had no such color change and have no significant influence on $Cu^{2+}$ recognition process. The interaction of $Cu^{2+}$ and sensor 1 was proven to adopt a 1:1 binding stoichiometry and the recognition process is reversible.

• 미생물학회지
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• 제30권4호
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• pp.305-309
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• 1992

Bacillus stearothermophilus 의 cytosine deaminase (EC 3. 5. 4.1 )를 수율 52.7% 로 7.2배 부분정제했다. 부분정제된 cytosine deaminase 는 cytosine 을 유일한 기질로 이용하였으머 5-methylcytosine 과 5-fluorocytosine 은 기질고 이용되지 못햇으며 cytosine 에 대한 Michaelis 정수 Km 값은 5.9 mM 이었다. 본효소는 pH 4.0 에서 7.0 까지의 폭 넓은 pH 영역에서 안정했으며 80.deg.C 에서 10 분간 열처리하여도 75% 이상의 효소활성이 잔존하여 높은 내열성 효소였다. 본 효소의 반응최적 pH 는 7.0-7.5 였으며 반응 최적 온도는 35. 37.deg.C 였다. 그리고 Arrhenium plot 에 의하여 계산된 활성화 에너지 값(Ea value) 은 26 Kcal/mol 이었다. 본 효소는 중금속인 1 mM의 $Cd^{2+}$ , $Hg^{2+}$ 및 $Cu^{2+}$ 에 의하여 완정히 실활되었으며 GMP 와 CMP 에 의해서는 효소활성이 촉진되었다. 특히 본 효소는 p-chloromercuribenzoate 에 의하여 효소 활성이 강하게 저해되어 thiol 효소임을 추정할 수 있었다.

• BMB Reports
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• 제28권2호
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• pp.170-176
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• 1995

Rhodanese from mouse liver was purified to near homogeneity by ammonium sulfate precipitation, CM-Sephadex ion exchange, hydroxyapatite and Sephacryl S-200-HR gel filtration chromatographies with a purification of 776 folds. The molecular weight was determined by Sephadex G-150 gel filtration and found to be 34.8 KDa. SOS-PAGE showed molecular weight 34 KDa and two identical subunits splitting by aging for 3 weeks at $-70^{\circ}C$ the molecular weight of which was 17 KDa. The optimal pH of enzyme activity was 9.4 and the pI value of the enzyme was 6.6. Rhodanese showed the optimal reaction temperature of $25^{\circ}C$ and near linear increasing pattern until 10 min. incubation. $K_m$ values of rhodanese for KCN and $Na_{2}S_{2}O_{3}$ as substrates were 12.5 mM and 8.3 mM, respectively. Rhodanese activity was inhibited by more than 70% at a concentration of 100 ${\mu}M$ of $Ni^{2+}$, $Zn^{2+}$, $Cd^{2+}$, $Hg^{2+}$ and $Cu^{2+}$. Other metal ions, such as $Mn^{2+}$, $Mg^{2-}$, $Ca^{2+}$, and $Fe^{2+}$ showed no effect on rhodanese activity.

• Journal of Microbiology and Biotechnology
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• 제20권7호
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• pp.1069-1076
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• 2010

A novel laccase from Tricholoma mongolicum was purified by using a procedure that entailed ion-exchange chromatographies on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and FPLC-gel filtration on Superdex 75. The purified enzyme was obtained with a specific activity of 1,480 U/mg-protein and a final yield of 15%. It was found to be a monomeric protein with a molecular mass of 66 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was GIGPVADLYVGNRIL, similar to some but also different to other mushroom laccases. The optimum pH and temperature for the purified enzyme were pH 2 to pH 3 and $30^{\circ}C$, respectively. It displayed a low $K_m$ toward 2,7-azinobis (3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) and high $k_{cat}/K_m$ values. The purified laccase oxidized a wide range of lignin-related phenols, but exerted maximal activity on ABTS. It was significantly inhibited by $Hg^{2+}$ ions, and remarkably stimulated by $Cu^{2+}$ ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma HepG2 cells and breast cancer MCF7 cells with an $IC_{50}$ of 0.65 ${\mu}M$, 1.4 ${\mu}M$, and 4.2 ${\mu}M$, respectively, indicating that it is also an antipathogenic protein.

• 한국균학회지
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• 제14권2호
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• pp.101-107
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• 1986

The properties of carboxyl proteinase which was contained in Poria cocos (Schw.) Wolf were investigated by means of the purification with 0.65 ammonium sulfate saturation, DEAE cellulose and Sephadex G-75 gel filtration. This enzyme was found to hydrolyze only peptide bond between glutamyl-L-tyrosine of carbobenzoxy-L-glutamyl-L-tyrosine among the synthetic substrates of carbobenzoxy-L-glutamyl-L-tyrosine, hippuryl- L-phenylalanine and hippuryl-L-arginine. This enzyme was inhibited by $Zn^{+2},\;Fe^{+2},\;Ca^{+2},\;CN^{-1},\;P_2O_7^{-4}$ ions, but stimulated by $Hg^{+2}$ ion. Also, this enzyme was inhibited by organic compounds such as L-lysine, L-phenylalanine, hippuryl-L-phenylalanine, diazoacetyl-DL-norleucine methyl ester (DAN) and 1,2-epoxy-3-(P-nitrophenoxy)propane(EPNP). In particular, the activity was inhibited by L-lysine till 20 minutes of preincubation time rapidly, and by DAN in the presence of $Cu^{+2}$ ion more rapidly after 30 minutes than DAN in the absence of $Cu^{+2}$ ion. L-Lysine was found to be a competitive inhibitor and its $K_i$ value was determined to be 0.12 mmole by Dixon plot.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.331-338
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• 2012

A novel gene coding for an endo-${\beta}$-1,4-mannanase (manA) from Bacillus subtilis strain G1 was cloned and overexpressed in P. pastoris GS115, and the enzyme was purified and characterized. The manA gene consisted of an open reading frame of 1,092 nucleotides, encoding a 364-aa protein, with a predicted molecular mass of 41 kDa. The ${\beta}$-mannanase showed an identity of 90.2-92.9% ${\leq}95%$) with the corresponding amino acid sequences from B. subtilis strains deposited in GenBank. The purified ${\beta}$-mannanase was a monomeric protein on SDS-PAGE with a specific activity of 2,718 U/mg and identified by MALDI-TOF mass spectrometry. The recombinant ${\beta}$-mannanase had an optimum temperature of $45^{\circ}C$ and optimum pH of 6.5. The enzyme was stable at temperatures up to $50^{\circ}C$ (for 8 h) and in the pH range of 5-9. EDTA and most tested metal ions showed a slightly to an obviously inhibitory effect on enzyme activity, whereas metal ions ($Hg^{2+}$, $Pb^{2+}$, and $Co^{2+}$) substantially inhibited the recombinant ${\beta}$-mannanase. The chemical additives including detergents (Triton X-100, Tween 20, and SDS) and organic solvents (methanol, ethanol, n-butanol, and acetone) decreased the enzyme activity, and especially no enzyme activity was observed by addition of SDS at the concentrations of 0.25-1.0% (w/v) or n-butanol at the concentrations of 20-30% (v/v). These results suggested that the ${\beta}$-mannanase expressed in P. pastoris could potentially be used as an additive in the feed for monogastric animals.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1138-1149
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• 2017

The use of microalgal biomass is an interesting technology for the removal of heavy metals from aqueous solutions owing to its high metal-binding capacity, but the interactions with bacteria as a strategy for the removal of toxic metals have been poorly studied. The goal of the current research was to investigate the potential of Burkholderia tropica co-immobilized with Chlorella sp. in polyurethane discs for the biosorption of Hg(II) from aqueous solutions and to evaluate the influence of different Hg(II) concentrations (0.041, 1.0, and 10 mg/l) and their exposure to different contact times corresponding to intervals of 1, 2, 4, 8, 16, and 32 h. As expected, microalgal bacterial biomass adhered and grew to form a biofilm on the support. The biosorption data followed pseudo-second-order kinetics, and the adsorption equilibrium was well described by either Langmuir or Freundlich adsorption isotherm, reaching equilibrium from 1 h. In both bacterial and microalgal immobilization systems in the co-immobilization of Chlorella sp. and B. tropica to different concentrations of Hg(II), the kinetics of biosorption of Hg(II) was significantly higher before 60 min of contact time. The highest percentage of biosorption of Hg(II) achieved in the co-immobilization system was 95% at pH 6.4, at 3.6 g of biosorbent, $30{\pm}1^{\circ}C$, and a mercury concentration of 1 mg/l before 60 min of contact time. This study showed that co-immobilization with B. tropica has synergistic effects on biosorption of Hg(II) ions and merits consideration in the design of future strategies for the removal of toxic metals.

• 한국식품영양과학회지
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• 제13권1호
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• pp.22-26
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• 1984

최근(最近) 산업(産業)의 발전(發展)과 더불어 매연(煤煙)이나 발수(廢水)에서 배출(排出)되어 나오는 유기금속화합물유(有機金屬化合物類)는 더욱 더 증가(增加)하여, 직접, 간접으로 우려의 환경(環境)을 오염(汚染)시켜, 인명(人命), 가축(家畜), 식물(植物) 및 수자원(水資源)에까지 심각한 피해(被害)를 끼치고 있다. 그리하여 본실험(本實驗)에서는 금속화합물중(金屬化合物中) 특(特)히 수은(水銀)의 급여(給與)에 의한 급만성중독시(急慢性中毒時) 나타나는 체내(體內) 물질대사(物質代謝)의 영향(影響)에 관한 검사(檢討)를 목적(目的)으로 증류수에 $HgCl_2$를 용해(溶解)시켜 Hg의 함량(含量)을 달리한 후 (0, 5, 40, 200 ppm) 흰쥐에게 80일간(日間) 급여(給與)하였을 때 다음과 같은 결과(結果)를 얻었다. 1. 체중증가율(體重增加率)은 수은(水銀) 함량(含量)이 높아질수록 현저히 저하(低下)되었으며 또한 각종(各種) 장기(臟器)의 중량(重量)은 200 ppm 군(群)에서 전장기(全臟器)가 유의성(有意性) 있는 증가(增加)를 나타내었다. 2. 혈액(血液)의 Hematocrit치(値)와 혈청(血淸)중의 GOT, GPT는 별다른 변화(變化)를 보이지 않았으나 cholesterol은 200 ppm군(群)에서 유의(有意)하게 증가(增加)되었고 총단백질(總蛋白質)과 albumin함량(含量) 및 A/G ratio도 200 ppm군(群)에서 유의적(有意的)인 감소(滅少)를 관찰(觀祭)할 수 있었다. 3. 간장(肝臟) 및 신장(腎臟)중의 Hg, Ca, Mg의 함량(含量)은 수은(水銀) 함량(含量)이 증가(增加)할수록 현저히 높아졌으나 Cu와 Zn은 수은(水銀) 함량(合量)이 증가(增加)할수록 간장(肝臟)에서는 다소 감소(減少)하였으나 신장(腎臟)에서는 오히려 증가(增加)하였다.

• Applied Biological Chemistry
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• 제13권1호
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• pp.87-92
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• 1970

본(本) Sericin 분해(分解) 효소(酵素) 강력(强力) 분필균주(分泌菌株) $S_4-1-1$이 생성(生成)하는 Sericinase 는 1. 열(熱)에 대(對)해 안정(安定)하여 $5^{\circ}C$에서 1개월간(個月間) $20^{\circ}C$에서 약(約) 30시간(時間)까지 그 활성(活性)이 지속(持續)되며 2. Fibroin 물질(物質)에 대(對)한 분해력(分解力)은 전연(全然) 인정(認定)되지 않았고 3. EDTA 및 $Ag^{++}$, $Hg^{++}$에 의(依)해 거의 완전실활(完全失活)되며 $Cu^{++}$, $Cd^{++}$에 의(依)해서도 비교적(比較的) 강(强)하게 실활(失活)되였다. 4. 계면활성제(界面活性劑) Peretex N의 혼존시(混存時) 그 활성(活性)이 다소(多少) 촉진(促進)되었다. 이와 같은 Sericin분해효소(分解酵素)를 가잠견(家蠶繭)의 해서(解舒)에 이용(利用)하므로서 현행(現行)되고 있는 자견(煮繭)에 비(比)해 1. 조사성적(繰絲成績) 즉 해서율(解舒率), 생사량비율등(生絲量比率等)이 오히려 우수(優秀)했으며 2, 사조반(絲條班), 소절(小節), 인장강도(引長强度)의 성적(成績) 역시(亦是) 자견(煮繭)에 비(比)해 하등(下等)의 손색(遜色)이 없었다.

• Journal of Microbiology and Biotechnology
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• 제19권2호
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• pp.187-193
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• 2009

A metagenomic library of surface-water microbes from the Yangtze River in China was constructed, and a novel esterase, designated as EstY, was isolated and characterized. EstY had 423 amino acids with an estimated molecular mass of 44 kDa and pI of 7.28. It hydrolyzed various p-nitrophenyl esters(acetate, butyrate, caprate, caprylate, laurate, myristate, and palmitate) and its best substrate was p-nitrophenyl caprate(C8). The optimum pH for EstY activity was 9.0 and the optimum temperature was $50^{\circ}C$. Metal ions, such as $Mn^{2+},\;Co^{2+},\;Hg^{2+},\;Zn^{2+},\;and\;Fe^{3+}$, strongly inhibited the activity of EstY, whereas $Mg^{2+}$ was required for maximal activity. Activity remained in the presence of 10% alcohol, acetone, isopropanol, and dimethyl sulfoxide, respectively. An analysis of the amino acid sequence deduced from estY revealed that it had 7 closely related lipolytic enzymes. Moreover, a sequence analysis showed that EstY, like its 7 relatives, did not belong to any known lipolytic enzyme family.