• 생약학회지
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• 제22권2호
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• pp.101-111
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• 1991

Polyphenol oxidase(PPO) was purified from an extract of Puerariae Radix by ammonium sulfate fractionation followed by Sephadex G-150 column chromatography, which resulted in a 56-fold increase in specific activity. The enzyme was optimum of pH 6.5. The optimum temperature of enzymic reaction was about $40^{\circ}$. The enzyme was thermostable with a half-life equal to 32 min at $70^{\circ}$. Km values of the PPO for catechol and pyrogallol from Lineweaver Burk plots were $1.3{\times}10^{-2}M$, $1.16{\times}10^{-2}M$, respectively. The substrate specificity of the Puerariae Radix PPO showed high affinity toward pyrogallol. Reducing reagents such as cysteine, potassium metabisulfite, ascorbic acid, 2-mercaptoethanol completely inhibited the PPO activity at $10^{-2}M$ level. Linewear-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, 4-nitrocatechol, potassium cyanide was competitive with Ki values of $4.3{\times10^{-2}M,\;0.73{\times}10^{-6}M,\;6.9{\times}10^{-6}M,\;6.4{\times}10^{-7}M$, respectively. The browning reaction by PPO was observed to decrease temporarily with the addition of sodium diethyl dithiocarbamate, a well known copper chelating agent. Among the divalent cations, $Cu^{2+}$ ion was strong activator on PPO and $Mn^{2+},\;Co^{2+}$ ions was effect on PPO activity. $Zn^{2+},\;Mg^{2+}$ ions was inhibitor on PPO.

• Membrane and Water Treatment
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• 제11권1호
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• pp.59-68
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• 2020

The study objective was to evaluate the enhanced removal of high concentrations of phosphorus from synthetic wastewater (solely phosphorus-containing) and real wastewater (pig manure) by using carbon nanotube (CNT)-coated steel slag. Generally, phosphorus removal by steel slag is attributed to Ca²⁺ eluted from the slag. However, in this study, CNT was used to control the excess release of Ca²⁺ from steel slag and increase the phosphorus removal. The phosphorus removal rate by the uncoated steel slag was lower than that of the CNT-coated steel slag, even though the Ca²⁺ concentrations were higher in the solution containing the uncoated steel slag. Therefore, the phosphorus removal could be attributed to both precipitation with Ca²⁺ eluted from steel slag in aqueous solution and adsorption onto the surface of the CNT-coated steel slag. Furthermore, the protons released from the CNT surface by exchanging with divalent cations acted to reduce the pH increase of the solution, which is attributed to the OH^- eluted from the steel slag. The adsorption isotherm and kinetics of the CNT-coated steel slags followed the Freundlich isotherm and pseudo-second-order model, respectively. The maximum adsorption capacity of the uncoated and CNT-coated steel slags was 6.127 and 9.268 mg P g^-1 slag, respectively. In addition, phosphorus from pig manure was more effectively removed by the CNT-coated steel slag than by the uncoated slag. Over 24 hours, the PO₄-P removal in pig manure was 12.3% higher by the CNT-coated slag. This CNT-coated steel slag can be used to remove both phosphorus and metals and has potential applications in high phosphorus-containing wastewater like pig manure.

• The Korean Journal of Physiology and Pharmacology
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• 제3권2호
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• pp.199-205
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• 1999

Vibrio vulnificus cytolysin caused platelet cytolysis and increased intracellular calcium concentration $([Ca^{2+}]_i)$ of rat platelets in a concentration-dependent manner. In the presence of V. vulnificus cytolysin (3 HU/ml), lactate dehydrogenase (LDH) activity was increased from $1.3{\pm}0.4%$ of control to $64.3{\pm}3.4%$ in platelet suspension buffer. In $Ca^{2+}-free$ platelet suspension buffer, however, V. vulnificus cytolysin did not induce $[Ca^{2+}]_i$ increase and LDH release. Addition of EGTA (2 mM) to suspension buffer after the initial $Ca^{2+}$ influx reversed $[Ca^{2+}]_i$ to the control level. However, a $Ca^{2+}$ channel blocker verapamil $(20\;{\mu}M)$ or mefenamic acid $(20\;{\mu}M)$ did not inhibit V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. Divalent cations such as $Co^{2+},\;Cd^{2+}\;or\;Mn^{2+}$ (2 mM each) also did not alter V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. V. vulnificus cytolysin (3 HU/ml)-induced calcium influx was completely blocked by lanthanum (2 mM). Lanthanum (2 mM) also completely blocked V. vulnificus cytolysin (3 HU/ml)-induced LDH release. Osmotic protectants such as, raffinose, sucrose or PEG600 (50 mM each) did not inhibit the lytic activity of V. vulnificus cytolysin. In conclusion, lanthanum sensitive $Ca^{2+}$ influx plays a significant role in Vibrio vulnificus cytolysin-induced platelet cytolysis and thrombocytopenia in V. vulnificus infection.

• 생약학회지
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• 제30권3호
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• pp.306-313
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• 1999

Polyphenol Oxidase(PPO) was purified from an extract of Ginkgo biloba leaves by ammonium sulfate fractionation followed by sephadex G-150 column chromatography, which resulted in a 18-fold increase in specific activity. The enzyme was most active at pH 8.5 and the temperature optimum for the PPO catechol oxidation reaction was $45^{\circ}C$. Heat inactivation studies showed that heating for 7, 9 and 48 min, at 80, 70 and $60^{\circ}C$ respectively caused a 50% loss in enzymatic activity and that the enzyme was completely inactivated after heat treatment at $90^{\circ}C$ for 60 min. Km values of the PPO for catechol, hydroquinone and 4-methylcatechol derived from Lineweaver-Burk plots were $6.06\;{\times}\;10^{-4}M,\;1.02\;{\times}\;10^{-3}M,\;1.41\;{\times}\;10^{-3}M$ respectively. Of the substrates tested, 4-methylcatechol was oxidized most readily and the enzyme did not oxidize monophenols. The enzyme datalyzed browning reaction was completely inhibited in the presence of reducing reagents, namely ascorbic acid, cysteine, glutathione, 2-mercaptoethanol, potassium metabisulfite at 0.5 mM level. Sodium chloride showed very little inhibition effect on Ginkgo biloba leaves PPO. Lineweaver-Burk analysis of inhibition data revealed that the inhibition by cysteine, 2-mercaptoethanol, potassium cyanide was competitive with ki values of $1.1\;{\times}\;10^{-5}M,\;2.4\;{\times}\;10^{-5}M,\;8\;{\times}\;10^{-5}M$, respectively. Among the divalent cations, $Cu^{2+}ion$ was a strong activator on PPO and $Mn^{2+}ion$ was little or no effect on PPO activity $Ni^{2+}ion$ was an inhibitor on PPO.

• Molecules and Cells
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• 제40권10호
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• pp.787-795
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• 2017

Avoiding ingestion of excessively salty food is essential for cation homeostasis that underlies various physiological processes in organisms. The molecular and cellular basis of the aversive salt taste, however, remains elusive. Through a behavioral reverse genetic screening, we discover that feeding suppression by $Na^+$-rich food requires Ionotropic Receptor 76b (Ir76b) in Drosophila labellar gustatory receptor neurons (GRNs). Concentrated sodium solutions with various anions caused feeding suppression dependent on Ir76b. Feeding aversion to caffeine and high concentrations of divalent cations and sorbitol was unimpaired in Ir76b-deficient animals, indicating sensory specificity of Ir76b-dependent $Na^+$ detection and the irrelevance of hyperosmolarity-driven mechanosensation to Ir76b-mediated feeding aversion. Ir76b-dependent $Na^+$-sensing GRNs in both L- and s-bristles are required for repulsion as opposed to the previous report where the L-bristle GRNs direct only low-$Na^+$ attraction. Our work extends the physiological implications of Ir76b from low-$Na^+$ attraction to high-$Na^+$ aversion, prompting further investigation of the physiological mechanisms that modulate two competing components of $Na^+$-evoked gustation coded in heterogeneous Ir76b-positive GRNs.

• 한국식품과학회지
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• 제31권2호
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• pp.522-526
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• 1999

S. mutans에 대한 키토산의 항균효과는 그 분자량에 따라 뚜렷한 차이를 보였으며, 약 30,000의 분자량을 갖는 키토산이 가장 높은 항균활성을 보였다. 키토산 처리에 의해 세포로부터 단백질, 핵산, 2가 양이온 등이 상당량 유출되는 것이 관찰되었으며, 세포내 물질의 유출정도는 키토산의 항균효과와 밀접한 상관성을 보였다. 이러한 세포내 물질의 유출은 양이온성을 띠는 키토산의 아민기와 세포벽내의 음이온 성분과의 강한 결합에 의해 세포벽의 결합력이 약화되면서 생기는 결과로서 추측되었다. 키토산처리에 의해 S. mutans의 세포벽이 원형질로부터 분리되고 세포내 물질이 세포벽 내에서 밖으로 유출되는 현상은 투과전자현미경(TEM)에 의해 확인되었다.

• 한국동물학회지
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• 제27권4호
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• pp.199-212
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• 1984

대장균의 세포질내에 존재하는 단백질 분해효소(protease Ci)를 추출하여 그 성질을 조사하였다. 이 효소는 insulin, glucagon 및 bovine growth hormone을 분해시키나, bovine serum albumin, casein 및 globin 등은 분해시키지 않는다. 이 효소의 native 한 상태에서의 분자량은 120,000이며, 54,000 dalton의 동일한 2개의 subunit로 구성되어 있다. Protease Ci의 최적 pH는 7.5이며 등전점은 5.5이다. 이 효소는 o-phenanthroline에 의해 저해되며, $Mn^++나 Co^++$와 같은 metal 이온들에 의하여 활성화되므로 metalloprotease임을 알 수 있다. 이 효소는 p-hydroxymercuribenzoate에 의해서 크게 저해되나 sulfhydryl protease의 specific한 저해제인 Ep475와 leupeptin에 의해서는 영향을 받지 않는다. 대장균 세포내에서는 또 다른 insulin 분해효소 (protease Pi)가 존재하는데, 이 효소는 periplasm에 존재하므로 protease Ci와는 다르다.

• Bulletin of the Korean Chemical Society
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• 제27권12호
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• pp.2002-2010
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• 2006

This study examined the quenching of ofloxacin (OFL) and flumequine (FLU) fluorescence by $Cuj^{2+}$, $Ni^{2+}$, $Co^{2+}$ and $Mn^{2+}$ in an aqueous solution. The change in the fluorescence intensity and lifetime was measured at various temperatures as a function of the quencher concentration. According to the Stern-Volmer plots, the fluorescence emission was quenched by both collisions (dynamic quenching) and complex formation (static quenching) with the same quencher but the effect of static quenching was larger than that of dynamic quenching. Large static and dynamic quenching constants for both OFL and FLU support significant ion-dipole and orbital-orbital interactions between fluorophore and quencher. For both molecules, the static and dynamic quenching constants by $Cu^{2+}$ were the largest among all the metal quenchers examined in this study. In addition, both the static and dynamic quenching mechanisms by $Cu^{2+}$ were somewhat different from the quenching caused by other metals. Between $Ni^{2+}$ and FLU, a different form of chemical interaction was observed compared with the interaction by other metals. The change in the absorption spectra as a result of the addition of a quencher provided information on static quenching. With all these metals, the static quenching constant of FLU was larger than those of OFL. The fluorescence of OFL was quite insensitive to both the dynamic and static quenching compared with FLU. This property of OFL can be explained by the twisted intramolecular charge transfer in the excited state.

• BMB Reports
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• 제37권4호
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• pp.466-473
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• 2004

A new acid deoxyribonuclease (DNase) was purified from the cultured mycelia of Cordyceps sinensis, and designated CSDNase. CSDNase was purified by $(NH_4)_2SO_4$ precipitation, Sephacryl S-100 HR gel filtration, weak anion-exchange HPLC, and gel filtration HPLC. The protein was single-chained, with an apparent molecular mass of ca. 34 kDa, as revealed by SDS-PAGE, and an isoelectric point of 7.05, as estimated by isoelectric focusing. CSDNase acted on both double-stranded (ds) and single- stranded (ss) DNA, but preferentially on dsDNA. The optimum pH of CSDNase was pH 5.5 and its optimum temperature 55. The activity of CSDNase was not dependent on divalent cations, but its enzymic activity was inhibited by high concentration of the cation: $MgCl_2$ above 150 mM, $MnCl_2$ above 200 mM, $ZnCl_2$ above 150 mM, $CaCl_2$ above 200 mM, NaCl above 300 mM, and KCl above 300 mM. CSDNase was found to hydrolyze DNA, and to generate 3-phosphate and 5-OH termini. These results indicate that the nucleolytic properties of CSDNase are essentially the same as those of other well-characterized acid DNases, and that CSDNase is a member of the acid DNase family. To our knowledge, this is the first report of an acid DNase in a fungus.

• 한국결정학회지
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• 제1권2호
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• pp.76-83
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• 1990

완전히 Ca2+ 이온으로 치환된 탈수구조(a=12 2f3(2) .41와 이 결정에 요오드가 흡착된 구조(a= 12.258(2) 입)를 21살에서 입방공간군 Pm3m를 사 용하여 단결정 X-선 회절법으로 구조를 해석하였다. 탈수구조는 I>3 w (I)인 206개의 반사를 사용하 여 Rw값이 0.081까지 정밀화되었고 요오드가 흡착 된 구조는 173개의 반사를 사용하여 Rw값이 0.084 까지 정밀화되었다. 각각의 구조에서 단위세포당 6 개의 Ca2+ 이온은 6-링의 산소와 결합하면서 3개 의 다른 3회 회전축상에 위치하였다. 탈수한 칼슘 치환 제올라이트 A는 80살에서 단위세포당 여섯개의 요오드 분자를 흡착하며 각 요오드 분자는 골조 산소 원자와 전하이동 착물을 형성하였다(I-0=3. 32(3) A, I-I=2.7l (2) A 그리고 I-I-0=180℃).