Metal-Catalyzed-Oxidation(MCO)系에 의한 蛋白質의 損傷에 대한 硏究

姜珍壎 高神大學校 保健科學硏究所 1992 보건과학연구소보 Vol.2 No.-

市販하는 各種 蛋白質과 生體中의 生化學的 代謝反應에 관여하는 酵素를 Cu(II)/EDTA/AsA의 反應系와 37℃에서 反應시키고 그 結果를 電氣泳動, HPLC 및 아미노산 분석기 등의 방법에 따라 조사하여 生體 高分子物質인 蛋白質과 酵素蛋白質에 대한 MCO系의 영향을 조사하였으며 金屬이온의 존재에 의하여 生成되는 각종 活性酵素의 영향을 조사하기 위하여 인위적으로 活性酵素의 生成系를 조제하여 BSA 및 酵素와 반응시킨 후 이들의 損傷作用을 측정하여 MCO系에 의한 蛋白質의 損傷機構를 構造的인 측면에서 밝히고자 하였다. 그 結果를 要約하면 다음과 같다. BSA와 milk casein은 Cu(II)/EDTA/AsA의 반응계에 의하여 크게 손상을 받는 것이 電氣泳動과 HPLC 및 아미노산 분석기 등의 기기실험을 통해 밝혀졌으며 蛋白質의 量的變化를 조사한 실험에서도 확인되었다. 2. BSA의 아미노산을 분석한 결과 histidine, arginine, tyrosine 및 phenylalanine 등의 아미노산이 消失되는 것을 알 수 있었으며 그 중에서도 histidine의 消失이 가장 큰 것으로 나타났다. 3. Glutathione peroxidase와 lactate dehydrogenase의 活性이 Cu(II)/EDATA/AsA의 반응계에 의하여 失活되는 것으로 나타났다. 4. Cu(II)에 의해서 뿐만 아니라 Fe(II)에 의해서도 BSA가 損傷을 받았으며 이 경우에도 histidine의 消失이 가장 크게 나타났으며 蛋白質의 損傷作用의 機構는 서로 달랐다. 5. 活性酵素의 生成系에 BSA를 반응시키고 아미노산 분석을 행한 결과 구성 아미노산의 대부분이 損傷을 받아 活性酵素種이 蛋白質의 損傷에 직접, 간접적으로 관여하는 것을 알 수 있었다. 6. 活性酵素의 生成系를 acetylcholine esterase와 반응시키고 그 活性의 低下를 조사한 結果 酵素의 失活이 큰 것으로 나타났으며 이 失活에는 過酸化水素, superoxide anion, 水酸라디칼 등의 活性酵素種이 크게 관여하는 것이 밝혀졌다. This study was carried out to elucidate on the effect of metal-catalyzed-oxidation system on the protein damage by reacting the protein(BSA, casein) and three enzymes, such as glutathione peroxidase, lactate dehydrogenase and acetylcholine esterase with Cu(II), or Fe(II)/EDTA/AsA system at 37℃, which results were obtained from the instrumental analysis through HPLC, SDS-polyacrylamide gel electrophoresis, amino acid analyzer and UV/VIS spectrum. Obtained results were as follows: 1. The proteins, such as BSA and casein, were greatly damaged by the system of Cu(II)/EDTA/AsA. 2. In the amino acid compositions, the concentrations of histidine, arginine, tyrosine and phenylalanine were decreased, among which the loss of histidine was about 60%. 3. The activity of glutathione peroxidase and lactate dehydrogenase was decreased by the system of Cu(II)/EDTA/AsA. 4. The system of Fe(II)/EDTA/AsA was also shown to damage the structure of BSA and acetylcholine esterase, through the experimental results using amino acid analyzer and UV/VIS spectrophotometer, and which results were due to the effects of active oxygen radical, such as ·OH, H₂O₂and superoxide anion(??) generated during the reaction. 5. In the alteration of the amino acid composition of BSA by reacting with Fe(II), the loss of histidine was greater than other amino acids. 6. The machanism of the protein damage by Cu(II)/EDTA/AsA was due to the oxidative degradation of the tertiary structure of the protein other than polymerization of the oxidized protein. 7. And, some active oxygens, such as hydrogen peroxide, superoxide anion and hydroxylradical, greatly affected the protein damage by Cu(II) or Fe(II)/EDTA/AsA system(MCO system).

市販하는 天然植物에서 由來하는 Flavonoid의 抗酸化能 및 DNA損傷抑制能에 관한 硏究

姜珍壎 高神大學校 保健科學硏究所 1994 보건과학연구소보 Vol.4 No.-

This study was carried out to elucidate on the inhibitiory effects on the lipid peroxidation and DNA damage of flavonoids in Natrural plants, using Leonuri Herba. Flavonoids obtained froom Leonuri Herba was fractionated by one large fraction and two small fractions, which were not fractionated correctly. HPLC chromatogram showed that each fraction obtained from Leonuri Herba was composed of a few flavonoids. All fractions had some antioxidative effect on the linoleic acid peroxidation, and also inhibitory effects on the DNA damage was shown to be in all fractions from experimental sample, but such effects were not so great.

고등어의 가공 및 저장중의 히스타민 생성에 미치는 첨가물의 영향(2) Glucose, Glycine, Sucrose 및 Sorbic Acid의 영향

강진훈,박영호,KANG Jin-Hoon,PARK Yeung-HO 한국수산과학회 1984 한국수산과학회지 Vol.17 No.6

적색육어류의 히스타만생성에 미치는 첨가물의 영향을 검토하기 위하여 고등어를 시료로 하여 이에 glucose, sucrose, glycine 및 sorbic acid 등을 첨가하여 저장하여 두고 저장기간에 따른 히스타민함량 및. 히스티딘탈탄산효소활성의 변화를 측정하였으며, 아울러 시료육의 성상이 히스타민생성에 미치는 영향을 조사하기 위하여 시료육의 역쇄정도를 달리 하였을 때의 히스타민함량의 변화를 비교, 검토하였다. 1. 균질화한 시료육에 glucose, sucrose 및 glycine을 각각 $5\%$ 및 $10\%$ 첨가하여 $25^{\circ}C$에 저장하였을 때 glucose 및 sucrose를 첨가한 것은 히스타민 생성을 다소 촉진하는 경향을 나타낸 반면, glycine을 첨가한 것은 히스타민생성 및 히스티딘탈탄산효소활성을 크게 억제하는 효과를 나타내어, $10\%$ 첨가한 것은 저장 5일 후에도 히스타민중독한계농도에 달하지 않았다. 2. 마쇄정도가 다른 시료육에 glucose 및 sucrose를 첨가하여 히스타민 생성에 미치는 영향을 조사한 결과 마쇄정도가 큰 것이 히스타민 생성을 촉진하는 경향을 나타내었다. 3. 시료육에 sorbic acid를 $0.1\%$ 및 $0.2\%$ 첨가하였을 때 히스타민 생성 및 히스티딘탈탄산효소활성은 크게 억제되었으며, $0.2\%$ 첨가한 것은 저장 6일째에 히스타민 함량이 최고치(275mg/100g)를 나타내었다. In succession to the previous paper, the effect of the addition of food additives, such as glucose, glycine, sucrose and sorbic acid on the histamine formation and histidine decarboxylase activity in mackerel muscle during storage at $25^{\circ}C$ was studied. Additionally, the effect of different rasping condition of mackerel muscle on the histamine formation was also studied. It was estimated that the rasping condition of mackerel muscle also affected to the histamine formation, by showing that much histamine was detected in homogenized muscle than in ground muscle when glucose and sucrose were added. The addition of glycine was inhibitory upon the histamine formation and histidine decarboxylase activity, which, in the muscle added $10\%$ of glycine, the histamine content was below the critical concentration of poisoning for histamine during storage for 5 days at $25^{\circ}C$. The addition of sorbic acid was also inhibited the histamine formation and histidine decarboxylase activity, and the inhibitory effect of $0.2\%$ addition was greater than $0.1\%$ addition.

過酸化脂質의 生成과 그 毒性에 대한 考察

姜珍壎 高神大學校 1988 論文集 Vol.16 No.-

[P8-91] 녹차와 둥굴레차의 항산화성에 대한 항산화비타민의 영향

강진훈 한국식품영양과학회 2009 한국식품영양과학회 학술대회발표집 Vol.2009 No.11

市販하는 天然植物에서 얻은 熱水 및 Methanol 推出物의 抗酸化能 및 金屬錯體形成能 : 山査子抽出物의 效果 The Effects of Crataegy Fructus

姜珍壎 高神大學校 1996 論文集 Vol.23 No.-

Theis study was carried out to elucidate the antioxidative effect, metal-chelating ability and the radical-scavenging effect of the antioxidative extracts obtained from Crataegy Fructus. The results were summarized as followed; 1. Hot water and methanol extracts of Crataegy Fructus were determined to have two or three fractions and one big fraction, each other. These results were the same as in the chromatograms of HPLC. 2. Both extracts of Crataegy Fructus showed to have the metal chelating ability with ??. 3. Both extracts showed to have the inhibitory effects against linoleic acid peroxidation, which the effect was bigger in the methanol extract than in hot water extract. 4. Methanol extract of Crataegy Fructus appeared to have the radical-scavenging effect, while it was not the case of hot water extract.

活性酸素種의 脂質酸化能 및 DNA損傷能에 관한 硏究

姜珍壎 高神大學校 1991 論文集 Vol.19 No.-

This study was attempted to investigate on the effects of active oxygens on the lipid peroxidation and the DNA damage by reacting linoleic acid and plasmid pBR 322 DNA with active oxygens, such as superoxide anion, hydrogen peroxide, singlet oxygen and hydroxyl radical, directly, at 37℃. Active oxygens, such as superoxide anion and hydrogen peroxide was repidly generated in the early stage of linoleic acid peroxidation, And, all active oxygen used greatly promoted the linoleic acid peroxidation from the early stage of the incubation. Plasmid pBR 322 DNA was apperaed to be greatly damaged by active oxygen from the results of quantitative measurement of DNA damage and agarose gel electrophoretic analysis And, it was assumed that DNA damage was by the random breaking of DNA double strands.

감귤에서 분리한 Bioflavonoid의 抗酸化作用 및 DNA 損傷抑制에 관한 硏究

姜珍壎 高神大學校 保健科學硏究所 1993 보건과학연구소보 Vol.3 No.-

This study was carried out to elucidate on the inhibitory effects on the lipid peroxidation and DNA damage of bioflavonoid obtained from Citrus sinensis. Flavonoids obtained from Citrus sinensis were fractionated by four fractions, such as compound I, II, III and IV, but they won't identifiec correctly, when compared with some standard flavonoids. Four fractions had some antioxidative effect on the linoleic acid peroxidation, and also inhibitory effects on the DNA damage was shown to be in all fractions from experimented sample, but such effects were not so great.

Maillard 反應由來 低分子 카르보닐화합물의 DNA 損傷作用에 대한 活性酸素種의 ]역할

姜珍壎 高神大學校 1990 論文集 Vol.18 No.2

The role of the active oxygens on plasmid DNA by carbonyl compounds derived from Maillard reaction was investigated. Plasmid DNA extracted from E.coli Hb101 was reacted with carbonyl compounds, such as glyoxal, methyl glyoxal, dihydroxyacetone, diacetyl, glyceraldehyde, glycolaldehyde and furfural with and without the active oxygen scavengers at 37℃ for 6 hours, and then the degree of damage was determined through the 1% agarose gel electrophoresis. All of the carbonyl compounds except furfural showed the demage of DNA. Among these, glyoxal, methyl glyoxal and dihydroxyacetone markedly induced the damage of DNA. On the other hand. the DNA damage by the carbonyl compounds was greatly inhibited by catalase, superoxide dismutase and α-tocopherol. Accordingly, it is considered that the damage of DNA is due to active oxygen, such as singlet oxygen, hydrogen peroxide and superoxide anion generated during the autoxidation of carbonyl compounds.

해조 추출물의 항산화 효소의 활성에 미치는 영향

강진훈 고신대학교보건과학연구소 2003 보건과학연구소보 Vol.13 No.-

미역과 다시마를 가정에서 조리하는 보편적인 방법인 가열에 의하여 및 메탄올을 이용해서 용해, 농축시켜서 이들이 가질 것으로 예상되는 각종의 실험을 행하였다. 즉 미역과 다시마를 시료로 하여 이들을 열수로 가열 농축하고 또한 메탄올을 이용하여 마쇄, 농축하여 일정한 고형성분을 분리하여 지질산화 억제능 및 활성산소종에 의한 영향을 비롯하여 이들 작용에 대한 항산화성 효소들의 영향을 실험을 통해 확인하였다. 또한 이들이 가지는 항산화성 외에도 노화의 한 분야인 고혈압 저해 능과 항산화효소들의 영향 둥을 실험하였다. 실험 결과 양 추출물에서 지질의 산화를 크게 억제하는 것을 알 수 있었으며 특히 미역에서 다시마보다 항산화능이 크며 항산화효소의 활성화에도 크게 영향을 미친다는 것을 확인 할 수 있었다. 이 같은 지질산화 억제효과는 열수추출물에서 더욱 크게 나타났으며 항산화성 효소들이 지질산화억제에 직접적으로 관여할 수 있다는 사실을 확인할 수 있었다. 한편, 이들 추출물은 활성산소종에 의한 산화촉진 및 유발 기능을 사전에 미리 예방하는데 상당히 유효한 작용을 한다는 결과를 나타내었다. 그리고 미역과 다시마의 추출물에서는 ACE의 활성을 저해하는 결과를 나타내었으며 특히, 미역에서 및 열수추출물에서 그 활성이 더욱 강하게 나타났다. This study was carried out to elucidate the effect of seaweed extract on the antioxidative enzyme activity and other physiological utility of seaweeds. The results obtained were as follows; Seaweed extracts from sea mustard and sea tangle showed a great antioxidative activity and it becomes bigger in the case of antioxidative enzymes addition, which enzymes were SOD, catalase, glutathione peroxidase (GPx). And all the experimented samples inhibited the linoleic acid peroxidation by active oxygen radicals, and the inhibition ability was bigger in hot-water extracts than in methanol extracts. The extracts made the antioxidative enzyme activity stronger after reaction, in which the ability was bigger in hot-water extracts than methanol extracts. And it showed to have the ability to inhibit the ACE activity, especially stronger in hot-water extracts and in sea mustard extract.