마늘 식초 및 호박 식초에 관한 연구

금종화 한국식품영양학회 1999 韓國食品營養學會誌 Vol.12 No.5

5% 에탄을 용액에 마늘을 파쇄하여 10% 가하고 Acetobacter aceti 3281로 35℃에서 26일간 발효시켜서 마늘식초를 제조하였다. 호박식초는 호박술을 기질로 하여 Acetobacter aceti 3281로 35℃에서 26일간 발효시켜서 제조하였다. 마늘식초와 호박식초의 총당은 0.04 ㎎/ml 및 1.53 ㎎/m1, 환원당은 0.122 ㎎/ml 및 0.406 ㎎/ml, 비중은 1.001 및 1.004, 에탄올은 모두 0.06% 및 0.02%, 균체수는 8.52 및 8.48 CFU/ml, pH는 3.06 및 3.20, 산도는 4.98 및 5.02를 나타냈다. 기호도는 마늘식초는 2.7. 호박식초는 3.9를 나타냈다. The garlic vinegar brewed with 5% ethanol solution added 10% crushed garlic was fermented by Acetobactor aceti 3281 at 30℃ for 26 days. Pumpkin wine vinegar was made from acetic acid fermentation of pumpkin wine at 35℃ for 26 days. The garlic vinegar and pumkin wine vinegar contained 0.04㎎/ml and 1.53㎎/ml of total sugar, 0.122/㎎/ml and 0.406㎎/ml of reducing sugar, and 0.06 and 0.02% of ethanol. Specific gravity of garlic vinegar and pumkin wine vinegar was 1.0012 and 1.0015, repectively. Viable cell count of garlic vinegar and pumkin wine vinegar was 8.53 and 8.48CFU/ml, respectively. pH of garlic vinegar and pumkin vinegar was 3.06 and 3.20, respectively. Acidity of garlic vinegar and pumkin wine vinegar was 4.98 and 5.02, respectively. Sensory evaluation garlic of vinegar and pumkin wine vinegar was 2.7 and 3.9, respectively.

Aspergillus niger α-Galactosidase의 정제 및 성질

금종화,오만진,김찬조 한국산업미생물학회 1991 한국미생물·생명공학회지 Vol.19 No.5

Aspergillus niger가 생산하는 α-glactosidase의 효소학적 성질을 조사하기 위하여 시험균주를 밀기울 배양한 후 생성된 α-glactosidase를 염석, 이온교환 크로마토그래피 및 겔 여과 등의 방법으로 정제한 후 정제효소의 효소학적 성질을 검토하였다. Asp. niger를 밀기울 배지에서 30℃, 4일 배양했을 때 효소활성이 가장 높았으며 α-glactosidase는 황산암모늄 염석, DEAE-cellulose 및 DEAE-Sephadex A-50 이온교환 크로마토그래피, Sephadex G-150 겔 여과 등에 의하여 23.7배까지 정제되었으며 비활성이 1,220 U/㎎·protein, 수율 14%이었고 HPLC와 PAGE에 의해 순도가 확인되었다. 정제효소는 당 단백질로서 등전점은 4.6이었고 분자량이 28,000인 monomer 4개로 구성된 tetramer이었다. 정제효소의 최적작용 pH는 6.5, 최적작용온도는 40℃이었고 60℃에서 10분 처리시 46%의 잔존활성을 나타내었다. 정제효소는 stachyose보다 raffinose를 쉽게 분해하였고 PMPG에 대한 K_m값은 5.0mM, 활성화에너지는 8.515Cal/mole이었다. To elucidate enzymatic properties of α-glactosidase(EC 3.2.1.22) from Asp. niger, α-glactosidase from wheat bran culture was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. And then its enzymatic propeties were investigated. The highest level of α-glactosidase activity was obtained when Asp. niger was grown on wheat bran medium at 30℃ for 96 hours. The α-glactosidase was purified by 23.7 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Celluose and Sephadex A-50, and gel filtration on Sephadex G-150 and its specific activity was 1,229 Units/㎎ protein and the yield was 14% of the total activity of wheat bran culture. The purified α-glactosidase was found to be homogeneous by polyacrylamide gel electrophoresis and HPLC. The α-glactosidase was a tetrameric glycoprotein which consisted of identical subunits with molecular weight of 28,000 each by SDS-PAGE and isoelectric point was determined analytical isoelectric focusing to be pH 4.6. The optimal temperature and pH for the α-glactosidase activity were 40℃ and pH 6.5, respectively, and 54% of its activity was lost by heating at 60℃ for 10 mins. It was appeared to have higher affinty to raffinose than to stachyose. The K_m value and activation energy of α-glactosidase were 5.0 mM and 8.515㎉ per mole for p-nitrophenyl-α-D-galactopyranoside, respectively.

Aspergillus niger $\alpha$-Galactosidase의 정제 및 성질

금종화,오만진,김찬조 한국미생물 · 생명공학회 1991 한국미생물·생명공학회지 Vol.19 No.5

Aspergillus niger가 생산하는 $\alpha$-galactosidase의 효소학적 성질을 조사하기 위하여 시험균주를 밀기울 배양한 후 생성된 $\alpha$-galactosidase를 염석, 이온교환 크로마토그래피 및 겔 여과 등의 방법으로 정제한 후 정제효소의 효소학적 성질을 검토하였다. Asp. niger를 밀기울 배지에서 $30^{\circ}C$, 4일 배양했을 때 효소활성이 가장 높았으며 $\alpha$-galactosidase는 황산암모늄 염석, DEAE-cellulose 및 DEAE-Sephadex A-50 이온교환 크로마토그래피, sephadex G-150 겔 여과 등에 의하여 23.7배까지 정제되었으며 비활성이 1,229U/mg.protein, 수율 14이었고 HPLC와 PAGE에 의해 순도가 확인되었다. To elucidate enzymatic properties of a-glactosidase (EC 3.2.1.22) from Asp. niger, a-galactosidase from wheat bran culture was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. And then its enzymatic propeties were investigated. The highest level of $\alpha$-galactosidase activity was obtained when Asp. niger was grown on wheat bran medium at $30^{\circ}C$ for 96 hours. The $\alpha$-galactosidase was purified by 23.7 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Celluose and Sephadex A-50, and gel filtration on Sephadex G-150 and its specific activity was 1,229 Unitslmg protein and the yield was 14% of the total activity of wheat bran culture. The purified $\alpha$-galactosidase was found to be homogeneous by polyacrylamide gel electrophoresis and HPLC. The $\alpha$-galactosidase was a tetrameric glycoprotein which consisted of identical subunits with molelcular weight of 28,000 each by SDS-PAGE and isoelectric point was determined analytical isoelectric focusing to be pH 4.6. The optimal temperature and pH for the $\alpha$-galactosidase activity were $40^{\circ}C$ and pH 6.5, respectively, and 54% of its activity was lost by heating at $60^{\circ}C$ for 10 mins, It was appeared to have higher affinty to raffinose than to stachyose. The K, value and activation energy of $\alpha$-galactosidase were 5.0 mM and 8.515 Kcal per mole for p-nitrophenyl- $\alpha$--D-galactopyranoside, respectively.

과요오드산 산화당에 의한 효소의 안정성

금종화 한국식품영양학회 2001 韓國食品營養學會誌 Vol.14 No.3

NaIO_4-산화 전분당을 Bzcillus licheniformisα-아밀라아제와 반응시켜서 시프염기 형성으로 당단백질로 변형시켜서 안정성을 확인하였다. 100℃에서의 열안정성은 10분 뒤에, pH 9.7에서 변형한 효소>pH 8.0에서 변형한 효소>비변형 효소의 순으로 높았다. 그러나 변형 및 안정성에 α-cyclodextrin(α-CD)을 사용한 결과 큰 차이는 나지 않았다. pH 8.0에서 α-CD존재 하에 변형한 효소는pH8∼11의 알칼리 쪽에서 가장 높은 안정성을 나타냈으나 pH5∼7 사이에서는 다른 효소보다 낮았다. pH 9.7에서 변형하지 않은 효소는pH 5부터 pH 13까지 서서히 증가하였고 pH 9.7에서 α-CD존재 하의 효소는pH 5부터 7까지 증가하다가 그후 pH 13까지 서서히 감소하였다. α-CD 존재 하의 비변형 효소는 pH 7과 10에서 피크를 나타낸 다음 pH 12 이후에는 급격히 낮아졌다. 변형한 효소는 HPLC의 유출시간이 빨라져서 변형하지 않은 효소보다 분자량이 큰 것으로 나타났다. 분자량 크기는 비변헝 효소 < pH 8,0에서 변형할 효소 < pH 9.7에서 변형한 효소의 순으로 컸다. NaIO_4-oxidized soluble starch was added to Bacillus licheniformis α-amylase by Schiff base reaction to make glycoprotein. Thermal stability at 100℃, in 10 minutes, proved high in the order of enzyme modified at pH 9.7, enzyme modified at pH 8.0 and native enzyme, respectively. But when α-cyclodextrin(α-CD) was used to modification and stabilization, the result showed no big difference. Modified enzyme under α-CD at pH 8.0 shows highest stability in pH 8∼11, while low in the pH 5∼7, compared to the other enzyme. Native enzyme at pH 9.7 gradually increased up to pH 13 from pH 5, enzyme under α-CD at pH 9.7 increased from pH 5 to 7 and little by little decreased up to pH 13. Native enzyme under α-CD peaked pH 7 and pH 10, and lowered sharply after pH 12. As molecular weight became larger than native enzyme, HPLC retention time of modified enzyme quickened. The molecular weight proved large in the order of modified enzyme at 9.7, enzyme modified at pH 8.0, and native enzyme.

대두 ${\alpha}-galactosidase$의 정제 및 성질

금종화,오만진,김성렬,Keum, Jong-Hwa,Oh, Man-Jin,Kim, Seong-Yeol 한국응용생명화학회 1991 Applied Biological Chemistry (Appl Biol Chem) Vol.34 No.3

To elucidate enzymatic properties of ${\alpha}-galactosidase$ (EC 3, 2, 1, 22) from germinated soybean, changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined. ${\alpha}-Galactosidase$ from germinated soybean was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties was investigated. ${\alpha}-galactosidase$ activity of sobeam was maximized when it was germinated at $25^{\circ}C$ for 120 hour. Raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination, respectively. Soybean ${\alpha}-galactosidase$ was purified by 6.6 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50, and gel filtration on Sephadex G-150. Its specific activity was 825 Units/mg protein and the yield was 2.5% of the total activity of crude extracts. The purified ${\alpha}-galactosidase$ of soybean was found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. Isoelectric point of soybean ${\alpha}-galactosidase$ was determined analytical isoelectric focusing to be pH 4.8. The soybean ${\alpha}-galactosidase$ was monomeric and its molecular weight was estimated to be 30,000 by SDS-PAGE. The optimal temperature and pH for the soybeam ${\alpha}-galactosidase$ activity were $40^{\circ}C$ and pH 6.0 and 75% of its activity was lost by heating at $60^{\circ}C$ for 10 min. The enzyme was appeared to have higher affinity to raffinose than to stachyose. The Km value of soybean enzyme was 5.3 mM for ${\rho}-nitrophenyl-{\alpha}-D-galactopyranoside$ and the activation energy on PNPG was calculated to be 13.02 Kcal per mole. 대두 발아 과정 중의 ${\alpha}-galactosidase$를 추출하여 염석, 이온교환 크로마토그래피 및 겔 여과 등의 방법으로 정제한 후 정제효소의 효소학적 성질을 검토하였다. 대두 ${\alpha}-galactosidase$의 활성은 $25^{\circ}C$에서 120시간 발아시켰을 때 가장 높았으며, 대두 중의 raffinose는 96시간, stachyose는 120시간 발아시켰을 때 완전히 분해되었다. 대두 ${\alpha}-galactosidase$는 황산암모늄염석, DEAE-Cellulose 및 DEAE-Sephadex A-50 이온교환 크로마토그래피, Sephadex G-150 겔 여과 등에 의하여 비활성은 825U/mg protein으로써 6.6배까지 정제되었으며 수율은 2.5%이었고 HPLC와 PAGE에 의하여 순도를 확인하였다. 정제효소의 등전점은 pH 4.8이었고, 분자량은 30,000인 monomer이었으며 정제효소의 최적작용 PH는 6.0, 최적작용온도는 $40^{\circ}C$ 이었고, $60^{\circ}C$에서 10분 처리시 25%의 잔존 활성을 나타내었다. 정제효소는 stachyose보다 raffinose를 쉽게 분해하였으며 PNPG에 대한 Km값은 5.3 mM, 활성화 에너지는 13.02 cal/mole이었다.

Aspergillus niger ATCC 16513과 대두(Glycine max. L) $\alpha$-galactosidase의 kinetic 성질

금종화,이종수,신철승,Geum, Jong-Hwa,Lee, Jong-Su,Sin, Cheol-Seung 배재대학교 자연과학연구소 1992 自然科學論文集 Vol.5 No.1

This experiment was carried out to elucidate some kinetic properties of the $\alpha$-galactosidase which produced and purified from Aspergillus niger ATCC 16513 and soybean(Glycine max. L). The Km value of Asp. niger and soybean $\alpha$-galactosidase were 37.0mM and 50.0mM for raffinose and55.5mM and 55.5mM for stachyose, respectively. The activity of Asp. niger and soybean $\alpha$-galactosidase were inhibited by galactose. Among the amino acids in active sites of both Asp. niger and soybean $\alpha$-galactosidase, histidine was identified by chemical modification of diethyl pyrocarbonate. Number of amino acids residues per mole of Asp. niger and soybean $\alpha$-galactosidase were 902 and 286, respectively. Aspergillus niger ATCC 16513과 대두(Glycine max. L)의 정제 $\alpha$-galactosidase를 사용 하여 몇가지 이들의 kinetic성질을 조사 하였다. Asp. niger $\alpha$-galactosidase의 raffinose와 stachyose에 대한 Km값은 각각 37.0mM과 55.5mM, 대두 $\alpha$-galactosidase는 50.0mM과 55.5mM로서 PNPG보다 이들에 대한 친화성이 적었다. 또한 galactose는 ASP. niger와 대두 $\alpha$-galactosidase 모두의 활성을 저해 하였으나 2-mereaptoethanol과 L-cystene은 대두 $\alpha$-galatosidase의 활성만을 약간 저해 하였다. Asp. niger와 대두 $\alpha$-galactosidase의 활성에 관여하는 아미노산은 diethyl pyrocarbonate에 의한 화학수식에 의하여 histidine임이 확인 되었고 Asp. niger $\alpha$-galactosidase의 1mole당 아미노산 잔기수는 모두 902개, 대두$\alpha$-galactosidase는 286개 이었다.

大豆 및 Aspergillus niger α-galactosidase의 酵素學的 硏究

琴鍾和,吳萬鎭 충남대학교 농업과학연구소 1991 농업과학연구 Vol.18 No.1

To elucidate enzymatic properties of α-galactosidases (EC3, 2. 1. 22) from germinated soybean and Aspergillus niger changes in the enzyme activities and oligosaccharide contents during germination of soybean were determined and α-galactosidases from germinated soybean and wheat bran culture of Aspergillus niger were purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. Their chemical and enzymatic properties were investigated and the results obtained were summarized as follows : 1. α-Galactosidase activity of soybean was maximized when it was germinated at 25℃ for 120 hours. And raffinose and stachyose in soybean were decomposed completely after 96 hours and 120 hours of germination. respectively. 2. The highest level of α-Galactosidase activity was obtained when Aspergillus niger was grown on wheat bran medium at 30℃ for 96 hours. 3. Soybean α-galactosidase was purified by6.6 fold by ammonium slufate fractionation, ion exchange chromatography on DEAE-Cellulose and Sephadex A-50., and gel filtration on Sephadex G-150. Its specific activity was 825 units/㎎ protein and the yield was 2.5% of the total activity of crude extracts. 4. Aspergillus niger α-galactosidase was purified by 23.7 fold. Its specific activity was 1,929 units/㎎ protein and the yield was 14% of the total activity of wheat bran culture. 5. The purified α-galactosidases of soybean and. Aspergillus niger were found to be homogeneous by polyacrylamide gel electrophoresis and by HPLC. 6. Chemical properties of the purified α-galactosidases were : 1) The soybean α-galactosidase was monomeric and its molecular weight was estimated to be 30.000 by SDS-PAGE whereas the Aspergillus niger α-galactosidase was a tetrameric glycoprotein which consisted of identical subunits with molecular weight of 28,000 each. 2) Isoelectric points of α-galactosidases were determined analytical isoelectric focusing to be : pH 4.8 for the soybean enzyme; and pH 4.6 for the Aspergillus niger enzyme, respectively. 3) Among the amino acids in active sites of both soybean and Aspergillus niger α-galactosidases, histidine was identified by chemical modification of diethyl pyrocarbonate. 1 4) The activity of soybean α-galatosidase was inhibited by 2-mercaptoethanol and L-cysteine. 7. Enzymatic properties of the purified α-galatosidases were : 1) The optimal temperature and pH for the α-galactosidase activity were : 40℃ and pH 6.0 for the soybean enzyme : and 40℃ and pH 6.5 for the Aspergillus niger enzyme. 2) The soybean α-galactosidase was stable at the range of pH 5.5 to 6.5 and at temperature below 45℃, however 75% of its activity was lost by heating at 60℃ for 10 min. The Aspergillus niger α-galactosidase was stable at the range of pH 6.0 to 7.0 and at temperature below 45℃, but 55% of its activity was lost under the same condition. 3) No significant difference was found in substrate specificity between (he bean and enzymes and both appeared to have higher affinity to raffinose than to stachyose. The enzymes were inhibited by galactose. 4) The Km values of soybean and Aspergillus niger enzymes were 5.3mM and 5.0mM for p-nitrophenyl-α-D-galactopyranoside, 50.0mM and 37.0mM for raffinose. and 55.5mM and 55.5mM for stachyose, respectively. 5) The activation energy and Q_10 value on p-nitrophenyl-α-D-galactopyranoside were calculated to be : 13.024 ㎉ per mole and 2.0 for the soybean enzyme; and 8.515 Kcal per mole and 1.38 for the Aspergillus niger enzyme.

淸州道民의 食道腐蝕症에 대한 臨床的 觀察(第1報)

金鍾和,金大成,金鎭永,金弘基 대한이비인후과학회 1974 대한이비인후과학회지 두경부외과학 Vol.17 No.1

大豆요구르트 製造過程中의 成分變化

琴鍾和,吳萬鎭 충남대학교 농업과학연구소 1984 農業技術硏究報告 Vol.11 No.1

This experiment was carried out to obtain the fundamental data for development of digestibility and quality enhanced product of soy yogurt. Soy yogurt was processed from raw materials of soybean, defatted soybean and sprouted soybean which inoculated with Lactobacillus acidophilus and Bifidobacterium bifidum as a starter. Changes of chemical compositions, viable cell count and saccharides during processing were investigated including acceptibility of manufactured products. The results were summarized as follows; 1 . Defatted soy milk fermented with Lactobacillus acidophilus was slowed the greatest initial acid productivity and sprouted soy milk was showed the greatest growth of lactobacillus acidophilus. 2. Acid production was accelerated when 2% glucose was used in soy milk. 3. Addition of reconstituted skim milk in soy milk and defatted soy milk increased acid production but was not showed the effect in the sprouted soy milk. 4. Sprouting soybean, the contents of raffinose and stachyose were decreased but those of glucose was increased. 5. When soy milk was fermented with Lactobacillus acidophilus, the contents of raffinose and stachyose were decreased. 6. As a result of panel test, sprouted soy yogurt which was produced by addition of reconstituted skim milk of 10% showed the greatest flavor and tastes.

慢性副鼻洞炎에 있어서 鼻內所見의 診斷的 價値에 관한 考察

金鍾和 대한이비인후과학회 1971 대한이비인후과학회지 두경부외과학 Vol.14 No.3