Aspergillus niger SFN-416으로부터 생산한 Xylanase 2의 분리정제 및 특성

성찬기,이상원,박석규,손봉수 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6

Aspergillus niger SFN-416으로부터 생성된 xylanase Ⅱ를 분리·정제하여 특성을 조사하였다. Aspergillus niger SFN-416의 배양액을 ethanol(70%) 침전, (NH_4)_2SO_4(30~90%) 침전, Sephadex G-100 chromatography 및 DEAE-Sephacel chromatography 등의 정제과정을 거친 결과, specific activity가 38.2 units/ mg protein으로 4.3배 정제되었고, 정제효소의 최적 활성온도는 50℃였다. 최적 pH는 5.5이었고, pH 4.5~6.0 사이에서 비교적 안정하였다. 또한 금속이온에 대한 효소의 활성은 대부분 억제를 보였고, 특히 Hg^2+에서는 3.7%로 강하게 효소활성이 저해됨을 보였고, Fe^2+에서는 100%의 상대활성을 나타내었다. 정제 효소의 분자량은 SDS-PAGE에 의하여 42,000 daltons이었으며, 유기용매에 대한 활성은 10%의 methanol, ethanol, isopropanol 및 1-butanol에 대하여 모두 낮은 활성을 나타었다. Xylanase (EC 3.2.1.8) was purified approximately 4,3 fold from Aspergillus niger SFN-416 by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-Sephacel ion exchange chromatography. Molecular weight of the enzyme was approximately 42,000 daltons. The optimum pH and temperature of the enzyme activity were 5.5 and 50℃, respectively. The enzyme activity was enhanced by Fe^2+, and inhibited by Hg^2+. The activity was decreased by addition of methanol, ethanol, isopropanol and 1-butanol at a concentration of 10%(v/v).

생강 중에 존재하는 tyrosinase 의 정제 특성에 관한 연구

성찬기,조성희 ( Chan Ki Sung,Sung Hye Cho ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.6

Tyrosinase was purified from ginger (Zingiber officinale Rosc.) by ammonium sulfate precipitation, Sephadex G-25 and DEAF-Sephacel column chromatography. Ginger tyrosinase contains three isozymes with molecular weight of P₁, 27,000, P₂, 20,000 and P₃, 41,000 daltons. Amino acid residues were P₁, 245, P₂, 182 and P₃, 373. The enzyme has high substrate specificity on o-dihydroxyphenols and trihydroxyphenols. The K_m values for L-DOPA were P₁, 11.24, P₂, 9.62 and P₃, 8.47 mM. Sodium diethyldithiocarbamate, sodium cyanide and L-ascorbic acid inhibited the activity of enzyme. Sodium diethyldithiocarbamate inhibited tyrosinase activity noncompetitively. The K_m values for sodium diethyldithiocarbamate were P₁, 0.46, P₂, 0.60 and P₃, 0.67 mM. The pH of maximum enzyme activity was 6.5. The temperature of maximum enzyme activity was 55℃. The enzyme activity was increased by addition of Ca^(2+), Hg^(2+), Cu^(2+) and Mn^(2+) ions, but it was decreased in the presence of Fe^2+, Zn^(2+), Mg^(2+) and Ba^(2+) ions.

감으로부터 Tyrosinase 의 정제 및 성질

성찬기,조성희 ( Chan Ki Sung,Sung Hye Cho ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.1

Tyrosinase was purified from Diospyros kaki Thunb by ammonium sulfate precipitation, Sephadex G-25 gel filtration and DEAE-sephacel chromatography. The molecular weight of tyrosinase was approximately 25,000 daltons with 227 amino acid residues. The enzyme has high substrate specificity on (-)epicatechin and pyrogallol. The K_m value for L-DOPA was 11.8 mM. Sodium diethyldithiocarbamate, sodium cyanide and L-ascorbic acid inhibited the activity of enzyme. The pH of maximum enzyme activity was 7.0. The temperature of maximum enzyme activity was 50℃. The activity was enhanced by addition of Ca^(2+), Hg^(2+) and Mn^(2+) ions, but it was decreased in the presence of Mg^(2+), Ba^(2+), Zn^(2+) and Fe^(2+) ions.

Staphylococcus xylosus SC-22가 생산하는 lipase의 정제 및 특성

성찬기,갈상완,이상원,최영주 한국생명과학회 2001 생명과학회지 Vol.11 No.5

Staphylococcus xylosus SC-22으로 생성된 lipase을 분리. 정제하여 특성을 조사하였다. Staphylococcus xylosus SC-22의 배양액을 ammonium sulfate (30~80%), Sepadex G-100 및 DEAE-Sephacel chromatography의 정제과정을 거친 결과, specific activity가 756.6 units/mg protein으로 19.3배 정제도었으며 수율은 17.2%로 나타났다. 정제효소의 분자랴은 47kDa, 정제된 효소의 특성은 최적온도은 4$0^{\circ}C$, 최적 pH는 8.0이었으며, pH 안정성 범위는 pH 6.0~10.0부근에서 비교적 안정하였다. Alkaline lipase의 활성은 C $u^{2+}$와 P $b^{2+}$에 의해 완전히 저해되었으며, F $e^{3+}$ 에 의해 50% 효소활성이 저해되었으나, $Ca^{2+}$, $Mg^{2+}$, $Na^{+}$에 의해 저해를 받지 않았다. A bacterial strain SC-22 which produced alkaline lipase was isolated from salf-fermented shrimps. Strains SC-22 was identified as Staphylococcus xylosus. An alkaline lipase excreted by Staphylococcus xylosus SC-22 was purified by ammonium sulfate predipitation and column chromatography on Sephadex G-100 and DEAE-Sephace. The specific activity of purified lipase was 756U/mg of protein with 17.2% yield. The approximate molecular weight of the purified enzyme was 47 kDa. The partially purified lipase preparation had and optimum temperature of 4$0^{\circ}C$, an optimum pH of 8.0, and a stable of 5~10. Lipase activities were enhanced by salt ions such as $Ca^{2+}$, $Mg^{2+}$,N $a^{2+}$ while inhibited remarkably by heavy metal ions, C $u^{2+}$ and P $b^{2+}$.EX> 2+/.

Aspergillus neger SFN-416으로부터 생산한 Xylanase II의 분리정제 및 특성

성찬기,이상원,박석규,손봉수,Sung, Chan-Ki,Lee, Sang-Won,Park, Seok-Kyu,Shon, Bong-Soo 한국미생물 · 생명공학회 1996 한국미생물·생명공학회지 Vol.24 No.6

Xylanase (EC 3.2.1.8) was purified approximately 4.3 fold from Aspergillus niger SFN-416 by ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-Sephacel ion exchange chromatography. Molecular weight of the enzyme was approximately 42,000 daltons. The optimum pH and temperature of the enzyme activity were 5.5 and 50$\circ$C, respectively. The enzyme activity was enhanced by Fe$^{2+}$, and inhibited by Hg$^{2+}$. The activity was decreased by addition of methanol, ethanol, isopropanol and 1-butanol at a concentration of 10%(v/v).

Aspergillus niger SFN-416으로부터 생산한 β-Glucosidase의 정제 및 특성

성찬기,이상원,박석규,박정로,문일식 한국산업미생물학회 1997 한국미생물·생명공학회지 Vol.25 No.1

β-Glucosidase (EC 3.2.1.21) was purified from Aspergillus niger SFN-416 by a sequential process of ammonium sulfate precipitation, Sephadex G-100 and DEAE-Sephacel column chromatography. Molecular weight of the enzyme was 46,000 daltons. The K_m and V_max values for PNPG were 0.67 mM and 25 moles/㎖·min., respectively. The optimum pH and temperature of the enzyme activity were 3.5 and 58℃, respectively. the enzyme activity was decreased by addition of metal ions, and increased by addition of methanol, ethanol, isopropanol and 1-butanol at a concentration of 10% (v/v). Stability of the enzyme was increased by addition of isopropanol and 1-butanol at a concentraion of 10% (v/v).

생강 중에 존재하는 Tyrosinase의 정제 특성에 관한 연구

성찬기,조성희,Sung, Chan-Ki,Cho, Sung-Hye 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.6

생강으로부터 tyrosinase를 추출하여 ammonium sulfate에 의한 침전, Sephadex G-25와 DEAE-Sephacel column chromatography하여 isozyme $P_1$과 $P_2$ 그리고 $P_3$를 정제하였다. 정제된 Isozyme의 분자량은 SDS-PAGE와 HPLC에 의하여 $P_1$은 27,000으로 $P_2$는 20,000 그리고 $P_3$는 41,000 daltons으로 밝혀졌으며, 아미노산 조성은 $P_1$이 245로 $P_2$는 182 그리고 $P_3$는 373잔기로 이루어져 있었다. Isozyme $P_1$과 $P_2$ 그리고 $P_3$는 monohydroxyphenol류의 기질에 대하여 거의 활성이 없었으나, o-dihydroxyphenol류와 trihydroxyphenol류에서 높은 활성을 나타내었다. m-dihydroxyphenol류는 catechin에서 높은 활성을 보였지만 그 외에서는 거의 활성을 보이지 않았고, p-dihydroxyphenol류에 대해서도 활성이 없었다. 또 L-DOPA에 대한 $K_m$값은 $P_1$이 11.24로 $P_2$는 9.62 그리고 $P_3$는 8.47 mM이었고, $V_{max}$값은 $P_1$이 21.05로 $P_2$는 13.89 그리고 $P_3$는 5.65 nmol/ml min이었다. Tyrosinase isozyme 모두는 sodium diethyldithiocarbamate, sodium cyanide와 L-ascorbic acid 10 mM에서 100% 저해 되었다. Sodium diethyldithiocarbamate는 비경쟁적 저해를 보였으며, $P_1$과 $P_2$ 그리고 $P_3$에 대한 $K_m$값은 각각 0.46과 0.60 그리고 0.67 mM이었다. $P_1$과 $P_2$ 그리고 $P_3$ isozyme의 반응 최적 pH와 반응 최적 온도는 모두 pH 6.5와 $55^{\circ}C$ 였다. 금속이온 중 $Ca^{2+}$, $Hg^{2+}$와 $Cu^{2+}$ 그리고 $Mn^{2+}$ 모든 isozyme $P_1$과 $P_2$ 그리고 $P_3$의 활성을 증가시키는 반면 $Fe^{2+}$, $Zn^{2+}$, $Mg^{2+}$ 그리고 $Ba^{2+}$는 활성을 저해시켰다. Tyrosinase was purified from ginger (Zingiber officinale Rosc.) by ammonium sulfate precipitation, Sephadex G-25 and DEAF-Sephacel column chromatography. Ginger tyrosinase contains three isozymes with molecular weight of $P_1$, 27,000, $P_2$, 20,000 and $P_3$, 41,000 daltons. Amino acid residues were $P_1$, 245, $P_2$, 182 and $P_3$, 373. The enzyme has high substrate specificity on o-dihydroxyphenols and trihydroxyphenols. The $K_m$ values for L-DOPA were $P_1$, 11.24, $P_2$, 9.62 and $P_3$, 8.47 mM. Sodium diethyldithiocarbamate, sodium cyanide and L-ascorbic acid inhibited the activity of enzyme. Sodium diethyldithiocarbamate inhibited tyrosinase activity noncompetitively. The $K_i$ values for sodium diethyldithiocarbamate were $P_1$, 0.46, $P_2$, 0.60 and $P_3$, 0.67 mM. The pH of maximum enzyme activity was 6.5. The temperature of maximum enzyme activity was $55^{\circ}C$. The enzyme activity was increased by addition of $Ca^{2+}$, $Hg^{2+}$, $Cu^{2+}$ and $Mn^{2+}$ ions, but it was decreased in the presence of $Fe^{2+}$, $Zn^{2+}$, $Mg^{2+}$ and $Ba^{2+}$ ions.

감중에 존재하는 Tyrosinase의 정제 및 특성에 관한 연구 : Diospyros Kaki Thunb

박두천,성찬기 國立 昌原大學校 基礎科學硏究所 1993 基礎科學硏究所論文集 Vol.5 No.-

감으로부터 tyrosinase를 추출하여 ammonium sulfate에 의한 침전, Sephadex G-25와 DEAE-Sephacel column chromatography하여 isozyme P₁과 P₂그리고 P₃를 정제하였다. Isozyme P₁과 P₂그리고 P₃는 monohydroxypenol류의 기질에 대하여 거의 활성이 없었으나, o-dihydroxyphenol류와 trihydroxyphenol류 중 phrogallol에서 높은 활성을 나타내었다. m-dihydroxyphenol류는 catechin에서 높은 활성을 보였지만 그 외에서는 거의 활성을 보이지 않았고, p-dihydroxyphenol류에 대해서도 활성이 없었다. 또 L-DOPA에 대한 Km 값은 P₁이 9.1로 P₂는 11.2 그리고 P₃는 7.4mM이었고, Vmax 값은 P₁이 13.7로 P₂는 13.9 그리고 P₃는 10.0 nmoles/ml.min이었다. Tyrosinase isozyme 모두는 sodium diethyldithiocarbamate, sodium cyanide와 L-ascorbic acid가 10 mM에 대하여 100% 저해되었다. Sodium diethyldithiocarbamate는 비경쟁적 저해를 보였으며, P₁과 P₂그리고 P₃에 대한 Km값은 각각 25.0과 52.6 그리고 13.5mM이었다. P₁과 P₂그리고 P₃ isozyme의 반응 최적 pH와 반응 최적온도는 각각 pH7과 50℃였다. 금속이온중 Ca²+와 Hg²+와 Cu²+ 그리고 Mn²+는 모든 isozyme P₁과 P₂그리고 P₃의 활성을 증가 시키는 반면 Fe²+와 Zn²+와 Mg²+그리고Ba²+는 활성을 저해시켰다. Tyrosinase was purified from persimmon(Diospyros kaki Thunb) by ammonium sulfate precipitation, Sephadex G-25 and DEAE-Sephacel column chromatography. The enzyme has high substrate specificity on o-dihydroxyphenols and trihydroxyphenols. The Km values for L-DOPA were P₁;13.7, P₂;13.9 and P₃;10.0 nmoles/ml.min. Sodium diethyldithiocarbamate, sodium cyanide and L-ascorbic acid inhibited the activity of enzyme. Sodium diethyldithiocarbamate inhibited tyrosinase activity noncompetitively. The Km values for sodium diethyldithiocarbamate were P₁;25.0, P₂;52.6 and P₃;13.5mM. The pH of maximum enzyme activity was increased by addition of Ca2+, Hg2+, Cu2+ and Mn2+ ions, but it was decreased in the presence of Fe2+, Zn2+, Mg2+ and Ba2+ ions.

무우종자의 발아촉진에 관한 연구

조성희,성찬기 中央大學校 自然科學硏究所 1987 自然科學硏究所 論文集 Vol.1 No.-

The changes of some components in the cell and germination rate of Raphonus Sativus L. seeds that were treated with GA₃, KINETIN and NAA were observed. The results were follows: 1. The content of soluble protein was increased in homogenate and cytosolic fractions of GA₃, and KINETIN treated seeds compared with one of control. 2. The content of glucose was increased in homoginate and cytosoli fractions of GA₃and KINETIN treated seeds compared with one of control on the half and the first day. 3. The activity of trehalase was increased in homogenate and cytosolic fractions of seeds which treated with GA₃and KINETIN compared with one of control on the first day. 4. The ATPase did not changed in all fractions of the untreated and stimulant treated seeds, during germination stages. 5. The germination rate was remarkedly increased by continuous soak treatment of GA₃and KINETIN on the first and second days but in the case of NAA treatment, it was very smiliar to control.

산포도와 머루중의 Polyphenol Oxidase에 관한 연구¹

성찬기,박희중,임흥빈,조성희 中央大學校 基礎科學硏究所 1997 基礎科學硏究所 論文集 Vol.11 No.-

Polyphenol oxidase was partially purified from fresh vitis flexuosa and vitis amurensis and its characteristics were studied. the experimental results were summarized as follows. 1.The enzyme activities were increased during the fruit developement gradually in two sample. 2.The optimum pH and temperature for activity were 5.5 and 30℃, respectively. 3.The enzyme was active with o-phenols and trihydroxyphenol, but inactive toward m-diphenols and monophenols. 4.The ?? value of the enzymes was 15.4 mM in vitis flexuosa and 12.5 mM in vitis amurensis with catechol as substrate. 5.Inhibitors sudies indicated that sodium diethyl-dithiocarbamate, L-cysteine, sodium bisulfite, L-ascorbic acid and 2-mercaptoeyhanol were the most potent. 6.The enzyme activity increament by ?? was much better than those by the other metal ions.