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.

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).

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.

Aspergillus niger SFN - 416이 생산하는 Xylanase Ⅰ의 정제 및 특성

성찬기(Chan-Ki Sung),이상원(Sang-Won Lee),박석규(Seok-Kyu Park),전순실(Soon-Sil Chun) 한국식품영양과학회 1996 한국식품영양과학회지 Vol.25 No.4

Aspergillus niger SFN-416으로 부터 생성한 xylanase I를 분리ㆍ정제하여 특성을 조사하였다. Aspergillus niger SFN-416의 배양액을 ethanol(70%) 칭전 , (NH₄)₂-S0₄(30~90%) 침전, Sephadex G-100 chromatography 빛 DEAE-Sephacel ion chromatography 등의 정 제 과정을 거친 결과, 10.2배 정제되었고, 정제효소의 최적 활성온도는 50℃였다. 최적 pH는 3.5이었고, pH 안정성은 6.0 이상에서 활성이 급격히 감소하였다. 또한 금속이온에 대한 효소의 활성은 대부분 억제를 보였고, 특히 Hg^(2+)는 18.5%로 가장 낮은 상대활성을 보였지만, Fe^(2+)는 117.0%, Mn^(2+)는 129.9%로 오히려 효소활성이 증가되었다 . 정제 효소의 분자량은 SDS-PAGE 에 의하여 31,000 daltons이었으며 , 유기용매에 대한 활성과 안정성은 10%의 methanol, ethanol, isopropanol 및 1-butanol에 대하여 모두 낮은 활성을 나타내어 유기용매에는 안정하지 않는 것으로 생각된다. Xylanase(EC. 3. 2. 1. 8) was purified approximately 10.2 fold from Aspergillus niger SFN-416 by a sequential process of ammonium sulfate fractionation, Sephadex G-100 gel filtration and DEAE-Sephacel ion exchange chromatography. Molecular weight of the enzyme was approximately 31,000 daltons. The optimum pH and temperature of the enzyme activity were 3.5 and 50℃, respectively. The enzyme activity was enhanced by Fe^(2+) and Mn^(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).

지리오갈피의 이화학적 분석

김명석(Myung-Suk Kim),성찬기(Chan-Ki Sung),김홍출(Hong-Chul Kim),갈상완(Sang-Wan Gal),이상원(Sang-Won Lee) 한국생명과학회 2009 생명과학회지 Vol.19 No.12

지리오갈피(Acanthopanax chilsanensis)를 이용한 지역특산품을 개발할 목적으로 먼저 지리오갈피 뿌리, 줄기 및 열매 자체에 대한 이화학적 성분분석을 행하였다. 지리오갈피의 일반성분을 분석한 결과 뿌리, 줄기 및 열매의 조지방은 각각 2.09, 2.51 및 7.94%이었으며, 조단백질은 11.50, 7.18 및 10.17%로 나타났다. 그리고 조회분은 각각 11.07, 6.85 및 6.38%로 뿌리 중의 조회분이 줄기와 열매 중의 조회분 보다 약간 높게 나타났다. 환원당은 각각 18.90, 10.70 및 24.05 g/100 g으로 측정되었다. 색도를 측정한 결과 명도(L)는 줄기가, 적색도(a)는 열매가, 그리고 황색도(b)는 뿌리가 높은 것으로 나타났다. 유리당은 뿌리, 줄기, 열매 모두 fructose, glucose 및 sucrose의 순으로 높게 나타났다. Acanthoside-D 함량은 줄기(18.95 ㎎/100 g), 뿌리(8.10 ㎎/100 g) 및 열매(2.85 ㎎/100 g)의 순으로 높게 나타났다. 유리아미노산은 줄기가 955.26 ㎎/100 g으로 뿌리보다는 4.5배, 열매보다는 8.5배 높게 검출되었다. 향기성분 중에서는 α-pinene, β-pinene, 3-carene 및 D-limonene 등과 같은 천연향기 성분이 동정되었다. This study was conducted to investigate the physicochemical compositions in the root, stem and fruit of A. chilsanensis. The contents of crude fat were 2.09, 2.51 and 7.94%, and crude proteins were 11.50%, 7.18% and 10.17%, respectively. Crude ash levels were 11.07, 6.85 and 6.38%, respectively, and it was higher in root than stem or fruit. The contents of reducing sugar were 18.90, 10.70 and 24.05 g/100 g in the root, stem and fruit of A. chilsanensis. As a result of color measurement, L value (lightness) of stem, a value (redness) of fruit and b value (yellowness) of root were high, respectively. The content of free sugar was high in all root, stem and fruit, in order of fructose, glucose and sucrose. Acanthoside-D, the main factor of A. chilsanensis, was 18.95 ㎎/100 g in stem, 8.10 ㎎/100 g in root and 2.85 ㎎100 g in fruit. Free amino acid in stem was 955.26 ㎎/100 g, which was 4.5 times higher than in stem and 8.5 times higher than in fruit. Natural aromas were identified by GC/GC-MS. Natural aromas such as α-pinene, β-pinene, 3-carene and D-limonene were detected in A. chilsanensis.

Inhibitory Effects of Acanthopanax chiisanensis Ethanolic Extracts on FcεRI α Chain Expression

Sun-Yup Shim(심선엽),Chan-Ki Sung(성찬기),Sang-Won Lee(이상원),Young-Ju Choi(최영주),Hyeung-Rak Kim(김형락),Dae-Seok Byun(변대석) 한국생명과학회 2007 생명과학회지 Vol.17 No.11

호염기구와 비만세포는 FcεRI을 매개로 한 알러지 반응에 있어 효과세포로서 중요한 역할을 담당한다. 인간 유래 호염기구성 세포주 KU812F 세포의 FcεRI α chain 발현에 있어 지리 오갈피의 저해 효과에 대해 연구하였다. 지리 오갈피의 뿌리 및 줄기를 에탄올로 추출하여, FcεRI α chain 저해 활성 실험에 이용하였다. 세포 표면의 FcεRI α chain 발현량을 flow cytometry로 분석한 결과, 지리 오갈피 뿌리 및 줄기 추출물에서 세포표면의 FcεRI 발현을 억제하는 효과를 나타냈다. 또한 지리 오갈피 뿌리 및 줄기 추출물은 FcεRI α chain mRNA 발현을 감소시켰으며, FcεRI을 매개로 한 히스타민 유리를 감소시켰다. 이러한 결과는 지리 오갈피의 뿌리 및 줄기 추출물이 FcεRI α chain 발현의 저하 조절 및 히스타민과 같은 염증매개인자의 분비를 저하시킴으로서 항 알러지 활성을 갖는데 중요한 역할을 할 것으로 판단된다. Basophils and mast cells play an important role in FcεRI-mediated allergic reaction as effector cells. We studied the effects of Acanthopanax chiisanensis on FcεRI α chain expression in human basophilic KU812F cells. Ethanol extracts from root and stem of A. chiisanensis were tested for inhibitory effects of FcεRI α chain expression. The cell surface FcεRI α chain expression was examined by flow cytometric analysis. All of the extracts of A. chiisanensis reduced the cell surface FcεRI α chain expression. Furthermore, A. chiisanensis extracts caused a decrease in the level of FcεRI α chain mRNA level and FcεRI-mediated histamine release. These results suggest that root and stem extracts of A. chiisanensis play an important role in anti-allergic activity via down-regulation of FcεRI α chain expression and decrease in release of inflammatory mediator such as histamine.

Rhizopus japonicus와 Zymomonas mobilis의 혼합고정화 배양계에 의한 생전분으로부터 에탄올 생산

최수철(Soo-Cheol Choi),이상원(Sang-Won Lee),박석규(Seok-Kyu Park),성찬기(Chan-Ki Sung),손봉수(Bong-Soo Shan),성낙계(Nack-Kie Sung) 한국식품영양과학회 1996 한국식품영양과학회지 Vol.25 No.4

호기성의 Rhizopus japonicus와 협기성 Zymomonas mobilis로 구성된 혼합고정화 배양계(R-Z계)를 제조하고, 생전분으로부터 에탄을 생산에 응용하였다. R. japonicus를 고정화배양하므로서 액체배양에서 보다 2배 높은 glucose량을 얻었다. R-Z계의 에탄을 생산량은 1.67g/L(Y_(P/S), 0.094)이었지만, 배양 24시간째부터 산소공급을 억제한 R-Z 24계에서는 6.54g/L(Y_(P/S), 0.38)의 에탄올을 얻어 대조구의 약 4배를 향상시켰다. 회분배양에서는 5%의 기질 농도가 적당하였으며, 생산된 에탄올은 15.02g/L(Y_(P/S), 0.36)이었다. 2% 기질을 5회 첨가한 유가배양에서는 2%기질의 회분배양과 동등한 수율인 0.38을 얻었다. Ethanol production from raw starch was performed using the co-immobilized culture system of Rhizopus japonicus and Zymomonas mobilis(R-Z). Glucose production in immobilized R. japonicus culture was 2-fold higher than that in free cell culture. Ethanol production was 1.67g/L(Y_(P/S), 0.094) and 6.54g/L(Y_(P/S). 0.38) in R-Z and R-Z 24 culture system, respectively. R-Z system was modified and designated as R-Z 24 system by replacing cotton plug with silicon check valve after 24h fermentation with R-Z system. Optimal substrate concentration for ethanol production in batch culture was 5%(w/v) and ethanol concentration produced was 15.02g/L(Y_(P/S), 0.36). Ethanol yield(Y_(P/S), 0.38) in fed-batch culture of 5 times with 2%(w/v) substrate was equal to that in batch culture of 2% (w/v) substrate.

Xanthomonas sp. EPS - 1이 생산하는 다당류의 점도

손봉수(Bong-Soo Son),박석규(Seok-Kyu Park),이상원(Sang-Won Lee),성찬기(Chan-Ki Sung),서권일(Kwon-Il Seo) 한국식품영양과학회 1996 한국식품영양과학회지 Vol.25 No.1

Xanthomonas sp. EPS-1으로 부터 생산된 다당류 EPS-1용액의 농도가 0.04(g/dl)일 때, 비점도는 0.137, 환원점도는 3.425, 상대점도는 1.137, 고유점도는 3.209임을 알 수 있었다. 겉보기 점도는 온도상승에 따라 낮아졌으나 온도를 증가하였을 때는 서서히 증가하였다. 열처리 후에도 물성은 거의 변화지 않았으며, 응집성은 좋지 않았으나 locust bean gum과의 혼합효과는 우수하였다. At the 0.04%(w/v) exopolysaccharide EPS-1 produced from Xanthomonas sp. EPS-1, specific viscosity(η_(sp)), reduced viscosity(η_(red)), relative viscosity(η_(r)) and inherent viscosity(η_(inh)) were determined to be 0.137, 3.425, 1.137 and 3.209 respectively. Apparent viscosity(η_(app)) of exopolysaccharide EPS-1 was decreased by increasing temperature and gradually increased by decreasing temperature. After heat treatment of EPS-1 solution, rheological properties were not changed. EPS-1 solution was undesirable in flocculation, but its viscosity was increased to Cp 510 with addition of locust bean gum.

곰팡이와 응고제에 따른 발효두부의 품질특성

이승화(Seung-Hwa Lee),김용택(Yong-Taek Kim),손미예(Mi-Yae Shon),성찬기(Chan-Ki Sung),박석규(Seok-Kyu Park) 한국식품영양과학회 2001 한국식품영양과학회지 Vol.30 No.4

균주(Act. elegans, Rhi. oligosporus)와 응고제(CaCl₂, citric acid)를 달리하여 제조한 발효두부의 품질특성을 조사하였다. 발효두부의 수분과 조단백질은 발효시간이 증가할수록 빠른 속도로 감소하였으며, 조지방과 조회분은 약간 증가하였다. pH는 발효가 진행됨에 따라 서서히 증가하는 경향을 보였고, 총산은 감소하였으나 시험구간별 뚜렷한 차이점은 없는 것으로 나타났다. 환원당은 발효 7일째까지는 서서히 증가하였으나 그 후 14일째까지 급속히 증가하였고, 아미노태 질소 및 암모니아태 질소 역시 빠르게 증가하였다. 무기성분은 K>Ca>Mg>Na 등의 순으로 높게 나타났으며, 각 군별 뚜렷한 차이는 보이지 않았다. 결론적으로 발효도의 지표인 환원당과 아미노태 질소함량을 증가시키기 위해서는 단기발효(7일)에서는 Act. elegans가 Rhi. oligosporus보다 효과적이었고, 장기발효(14일)에서는 반대경향을 나타내었다. 응고제는 동일균주로 장기발효를 할 경우 CaCl₂가 citric acid 보다 효과적이었다. Changes of quality properties of fermented tofu prepared with two molds like Actinomucor elegans (AE) and Rhizopus oligosporus (RO) and coagulants (CaCl₂ and citric acid) were investigated. Moisture and crude protein of fermented tofu were rapidly decreased during fermentation, the contents of crude lipid and crude ash were shown to be slightly increased, and then total acidity was slowly decreased. The content of reducing sugar of fermented tofu was slowly increased for 7 day of fermentation, but rapidly increased after that time because of rapid hydrolysis of carbohydrate in fermented tofu. The contents of amino and ammonia type nitrogen were quickly increased during fermentation. The highest contents of amino type nitrogen of fermented tofu were found in sample of CaCl₂ group as a coagulant and RO group as a mold. Contents of minerals in tofu fermented for 14 day were high in order of K>Ca>Mg>Na. In conclusion, AE was more effective than RO to enhance the contents of reducing sugar and amino type nitrogen as an indicator of fermentation within 7 day of fermentation, and then RO was more effective than AE after that time. Calcium chloride as a coagulant was more effective than citric acid in tofu fermented with the same strain for 14 day.