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        부들 화분 혈전 용해효소의 정제와 특성

        박혜민(Hae-Min Park),구자형(Ja-Hyeong Gu),오만진(Man-Jin Oh) 충남대학교 농업과학연구소 2009 농업과학연구 Vol.36 No.1

        부들 화분(蒲黃)의 혈전 용해능을 검토하기 위하여 부들 화분을 물 추출하여 혈전 분해능이 있음을 확인하였다. 부들 화분의 혈전용해효소를 DEAE-cellulose, Sephadex G-150을 이용한 gel filteration, HPLC로 정제하여acrylamide gel electrophoresi로 정제를 하였다. 정제효소는 HPLC와 전기영동에 의하여 순수하게 정제되었음을 확인하였고 비활성은 38U/㎎로서 정제도는 86.4배이었고 분자량은 75kDa이었다. 정제효소의 최적 pH는 4.0이었고 pH 4.0-6.0에서 안정하였으며 정제효소의 최적온도는 55℃이었고 30-60℃에서는 안정하였으나 70℃ 부터 활성이 현저히 저하하여 80℃ 이상에서는 완전히 실활하였다. 기질특이성은 fibrin을 가장 잘 분해하였고 fibrin, whole casein, κ-casein, α-casein, β-casein, BSA순으로 나타났다. Casein을 기질로 하였을 때 Km value는 0.44 mM 이었으며 casein에 대한 친화력이 높은 것으로 나타났다. 효소활성에 미치는 금속이온의 영향은 K?, Na?, Mg<SUP>2+</SUP> 등은 효소반응에 영향을 미치지 않았으나 Zn<SUP>2+</SUP>, Fe<SUP>2+</SUP>는 심하게 저해작용을 나타내었고 정제효소는 PMSF, iodoacetic acid 및 SDS에 의하여 심하게 저해되는 것으로 보아 serine protease로 추정되었다. When the cattail pollen was identified by using fibrinolytic agents, we found that the fibrinolytic activity was controlled by an enzyme. Therefore, for determining the fibrinolytic activity of cattail pollen, the fibrinolytic enzyme in cattail pollen was purified by gel filtration using DEAE-cellulose, Sephadex G-150 and HPLC. Also, its purity was certified by polyacrylamide gel electrophoresis, and its physico-chemical properties, such as pH and temperature stabilities and effects of metal, inhibitors and substrates, were examined. The specific activity, purification fold, and molecular weight of the enzyme were 38U/mg, 86.4,and 75kDa, respectively. The optimum pH for the purified enzyme was at 4.0 and it was stable at pH 4.0-6.0. The optimum temperature was 55℃and it was stable at 30-60℃. But the enzyme began to be inactivated at 70℃ and its activity was totally lost at temperatures above 80℃. As for substrate specificity, the enzyme was most effective in dissolving fibrin, followed by whole casein, κ-casein, α-casein, β-casein, and BSA. With casein as the substrate, Km value was found to be 0.44mM and the enzyme showed a high affinity for casein. As for the metal ions affecting enzyme activity, K?, Na?, and Mg<SUP>2+</SUP> had no effect on enzyme reaction while Zn<SUP>2+</SUP> and Fe<SUP>2+</SUP> showed potent inhibitory activity. Judging from the fact that the purified enzyme was also strongly inhibited by PMSF, iodoacetic acid, and SDA, it assumed to be a serine protease.

      • KCI등재후보

        신고배(Pyrus pyrifolia Nakai)를 이용한 배술 제조 과정 중 성분 변화

        박혜민(Hae-Min Park),이가순(Ka-Soon Lee),홍종숙(Jong-Sook Hong),오만진(Man-Jin Oh) 충남대학교 농업과학연구소 2009 농업과학연구 Vol.36 No.2

        8종의 효모를 이용하여 담금한 후 배술의 pH, 산도, 에탄올 및 당도의 변화 및 기호도를 조사한 결과 사용된 효모 모두 발효가 지속되면서 일정기간까지 pH는 감소하였으며 산도는 증가하였고 에탄올함량은 발효 12일까지 증가하였고 당도는 12일까지 지속적으로 감소하였다. 효모의 종류에 따라 제조된 배술에 대한 기호도를 조사한 결과, 에탄올 생성량이 가장 높았던 처리구인 효모 B처리구에서 20-30대와 50-60대 연령층을 가진 관능요원 모두 기호도가 우수하였다. 또 효소제, 발효개선제 및 살균제의 첨가에 따른 배술의 화학적 특성을 조사한 결과, 각종 첨가제를 첨가함으로써 무 첨가구에 비하여 pH값이 낮아졌으며 산도는 ammonium phosphate 첨가구가 가장 높았다. 그러나 발효 4일 이후 산도의 변화는 거의 나타나지 않았다. 효소제, 발효개선제, 살균제를 첨가하여 담금한 술의 당도를 측정한 결과 K₂S₂O? 350ppm 첨가구가 가장 빠르게 당도가 감소하였으며 무 첨가구, ammonium phosphate, hemicellulase, pectinase, K₂S₂O? 175ppm순으로 나타났다. 6개의 실험구 모두 발효 과정 중 당도의 감소 속도는 큰 차이 없이 꾸준히 감소하였으며 발효 7일 이후 당도의 감소변화는 거의 없었다. 최종 당도는 9.5~9.8°Brix로 큰 차이는 나타나지 않았다. 에탄올 함량변화는 모든 실험구에서 발효 개시 7일 후 11~12.4%의 에탄올을 생성하였으며 ammonium phosphate을 첨가한 구가 12.4%로 알코올 생성속도가 가장 빨랐고 hemicellulase 첨가구는 전반적으로 알코올 생성속도가 늦었다. 메탄올 함량은 무 첨가구 및 K₂S₂O? 첨가구는 0.05%정도의 함량으로 거의 미비하게 검출되었으며 hemicellulase 및 pectinase 첨가구에서는 0.5%이상의 함량으로 다소 높은 함량을 보여주었다. For increasing the consumption of pear, the pear wines were made by using eight different kinds of yeast. The chemical properties, such as pH, acidity, ethanol and sugar contents, and the consumer acceptance of various pear wines were analyzed. In analysis result of chemical properties, the value of pH was decreased and the acidity was increased until few days, and then retained during fermentation. The ethanol content was increased and the sugar content was decreased during fermentation. The pear wine brewed by B yeast (Lalnin, Saccharomyces cerevisiae) among eight different kinds of yeast had the highest content of ethanol, also this wine was represented the most consumer acceptance by the panels who consist of two age groups such as 20's to 30's and 50's to 60's. Therefore, we chose the yeast B for analyzing the effect of additives during fermentation. As the additives, the K₂S₂O? and ammonium phosphate as the disinfectant and hemicellulase and pectinase as fermenting conditioner were added into the pear wine brewed by yeast B. When additives are added, the chemical properties were the same as followed. Te values of pH was lower than when additives were not added(control). The acidity was he highest when ammonium phosphate was added, but the acidity was not changed after 4 day fermentation. Sugar contents when added K₂S₂O? 350ppm was shown the fastest deceasing, and it was orderly decreased as control, ammonium phosphate, hemicellulase, pectinase, and K₂S₂O? 175ppm. The decreasing rate of sugar content of these 6 test groups during fermentation gradually decreased without a big changes and the after 7 day fermentation, sugar contents were seldom decreased and the final sugar content was 9.5~9.8°Brix. In terms of change of the content of ethanol, all test groups generated 11~12.4% ethanol after 7 day fermentation. The group where ammonium phosphate was added generated 12.4% alcohol, which is the fastest speed and the group, which hemicellulase additive was added, showed generally slow alcohol generation rate. In terms of ethanol contents, the non-additive group and K₂S₂O? additive group showed 0.05%, which is very low, and hemicellulase and pectinase additive group is more than 0.5% which is rather high.

      • KCI등재
      • KCI등재
      • KCI등재후보

        포황(蒲黃)의 성분분석과 생리활성

        이병찬(Bung-Chan Lee),박혜민(Hae-Min Park),심후성(Hu-Sung Sim),김곤섭(Gon-Sup Kim),구자형(Ja-Hyeong Gu),오만진(Man-jin Oh) 충남대학교 농업과학연구소 2009 농업과학연구 Vol.36 No.2

        포황을 기능성 식품의 소재로 이용하기 위한 가능성을 검토하기 위하여 일반 성분, 유리 당, 유리 아미노산, 지방산 조성, flavonoid 등을 분석하고 생리활성을 측정하였다. 포황의 일반성분은 수분 12.7-13.2%, 조단백질 15.7-17.8%, 조지방 1.3%, 유리당 7.5-7.7%, 조섬유 14.7-18.6%, 회분 3.4-4.9%, 가용성 무질소물 49.7-55.9%이었다. 유리아미노산 함량은 애기부들이 1.923 %, 좀 부들이 0.907 %, 큰 부들이 0.333 %로서 품종 간에 큰 차이를 나타내었고 그 조성은 모든 품종이 GABA alanine, glutamic acid, proline의 순이었다. 면역 활성을 높여주고 혈압강하 활성이 높다고 알려진 GABA함량이 50% 이상을 점하고 있어 매우 유용한 기능성 식품 소재로 판단된다. 지방산 조성은 linoleic, palmitic acid, oleic acid, linolenic acid 순으로서 품종 간에 큰 차이가 없었고 불포화 지방산의 비율이 67.2-76.0%로서 매우 높았다. 무기질 함량은 K이 0.354-0.492% 으로서 가장 높았으며 Mg 0.0516-0.0546 %, Ca 0.045-0.0486 %, Na 0.0101-0.0204 % 이었다. 항산화 물질로 알려진 포황 중의 flavonoid는 quercetin이 0.169-0.186 %로서 대부분을 차지하였고 rutin이 0.0094-0.0147 %이었다. SD계 rats에 4주간 포황과 그 추출물을 식이하면서 rats의 혈청에서 DPPH radical 소거활성을 알아 본 결과 소거활성 효과가 유의하게 나타났고, 혈중 cholesterol 함량을 낮추어 주는 효과와 혈중 glucose 함량을 낮추어 주는 결과로 보아 고지혈증의 예방과 당뇨병 치료에 효과가 있을 것으로 판단되었다. For utilizing Cattail pollen as a raw material for functional foods, the nutrients such as free sugar, free amino acid, fatty acid composition, flavonoid content, and the biological activity within Cattail pollen were measured. The results of proximate analysis within Cattail pollen included the following readings: 12.7-13.2% of moisture, 15.7-17.8% of crude protein, 1.3% of crude fat, 7.5-7.7% of free sugar, 13.7-18.6% of crude fiber, 3.4-4.9% of ash, and 49.7-55.9% of nitrogen free extracts. The composition of free amino acids consisted of 1.923% of T. orientalis, 0.907% of T. angustata, and 0.333% of T. latifolia, which were measurements that varied significantly among different species. However, all species showed considerable portions of GABA alanine, glutamic acid, and proline. Specifically, it was shown that the GABA composition, which is known for increasing immunity while simultaneously lowering blood pressure, exceeded 50%. Therefore, this result implies that Cattail pollens have potential as a powerful utilization for functional foods. The composition of the fatty acids mainly consisted of linoeic, palmitic acid, oleic acid, and linolenic acid, and didn’t show many variances across different species. Also, the total contents of unsaturated fatty acid were particularly high with a measured ratio of 67.2-76.0% value. Mineral in Cattail pollen was composed of 0.354-0.492% of K, 0.0516-0.0546% of Mg, 0.045-0.0486% of Ca, and 0.0101-0.0204% of Na. Among the Cattail pollens known as anti-oxidants, flavonoid contains 0.169-0.186% of quercetin, and therefore is the largest constituent followed by rutin making up a measurement of 0.0094-0.0147%. For the purpose of the study, the Cattail pollen and its extracts were fed to SC class rats for a span of 4 weeks. Then, the DPPH radical scavenging activity was measured from the tested rats’serums and the results showed significant variances. Also, the results indicated that the cholesterol and glucose levels in the blood were decreased which in turn led to the conclusion that the cattail pollen can help hyperlipidemia and diabetic treatments.

      • KCI등재

        불응성 용혈성 빈혈을 지닌 소아 전신홍반루푸스에서의 Rituximab 치료 1례

        박주환 ( Ju Hwan Park ),임재욱 ( Jae Wook Im ),전홍길 ( Hong Kil Jun ),박혜민 ( Hae Min Park ),최승원 ( Seung Won Choi ),박상규 ( Sang Kyu Park ),오지선 ( Ji Seon Oh ) 대한류마티스학회 2014 대한류마티스학회지 Vol.21 No.4

        Autoimmune hemolytic anemia (AIHA) is a relatively common cause of anemia in children and adults with systemic lupus erythematosus (SLE). Although AIHA responds to steroids, in case of refractory or steroid-dependent AIHA, immunosuppressants and intravenous immunoglobulin have been used as second line agents. Rituximab, an anti-CD20 monoclonal antibody, is emerging in the treatment of SLE refractory to conventional therapy. Herein, we report a case of delayed and sustained remission of refractory hemolytic anemia in a child with SLE, post rituximab treatment. A 12-year-old female child with dizziness was referred to our department and was diagnosed with SLE combined with hemolytic anemia and renal tubular acidosis. Since frequent relapse of hemolytic anemia had occurred during the steroid tapering course, even though she had been treated with additional immunosuppressants (azathioprine, mycophenolate mofetil), the patient received 2 doses of rituximab 500 mg at 2 weeks interval at 18 months post diagnosis. After 15 months of rituximab administration, her anemia and renal tubular acidosis were fully recovered, enough to stop all medications. She remained well without recurrence for up to 3 years and 4 months after rituximab treatment.

      • 균일 LED조명을 위한 자유형상 2차 광학계 설계 및 공차 분석

        양재석(Jae Suk Yang),박혜민(Hae Min Park),김대찬(Dae Chan Kim),오범환(Beom Hoan O),이승걸(Seung Gol Lee) 한국조명·전기설비학회 2011 한국조명·전기설비학회 학술대회논문집 Vol.2011 No.11

        A freeform lens was optimally designed for achieving an uniform illumination and also its tolerance analysis was performed by considering alignement and fabrication errores. Divergent model was chosen for establishing the ray-arriving point relation for uniform illumination, and the differential equation for the lens shape was numerically solved. The illuminance distribution on the target area was calculated with LightTools™ and the uniformity was extracted from the distribution. From the tolerance analysis, it was confirmed that the alignment error could give harmful effect on the performance.

      • KCI등재

        열처리 가공조건에 따른 씀바귀 침출차의 생리활성

        이가순(Ka-Soon Lee),김관후(Gwan-Hou Kim),김현호(Hyun-Ho Kim),김은수(Eun-Soo Kim),박혜민(Hae-Min Park),오만진(Man-Jin Oh) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.4

        씀바귀뿌리를 이용하여 침출차를 제조하고자 건조온도 및 처리조건을 달리하여 생리활성을 검토하였다. 각각의 씀바귀뿌리 침출차의 침출액에 대한 생리활성을 측정한 결과, 전자공여능은 증숙처리 후 열풍건조한 것이 98%로 가장 높았고 뒤이어 천일건조>열풍건조 후 볶음처리>열풍건조의 순으로 높았으며, 침출액의 SOD유사활성은 열풍건조 후 볶음 처리한 씀바귀뿌리 추출물에서 35.61±0.67%의 활성을 보였다. ACE저해활성은 천일건조한 것이 44.60%로 가장 저해활성이 가장 높았고, 증숙 처리 후 열풍건조한 처리구가 가장 낮아 28.89%의 저해활성을 보였고, tyrosinase 저해활성은 천일건조한 것이 44.61%로 가장 높았고 열풍건조>열풍건조 후 볶음처리>증숙처리 후 열풍건조의 순으로 저해활성을 나타내었으며, 아질산염 소거능은 pH가 낮을수록 효과가 있었으며, 특히 열풍건조 후 볶음처리를 한 씀바귀뿌리 침출차의 침출액이 모든 pH 구간에서 가장 효과적인 것을 볼 수 있었다. This study was carried out to investigate physiological functionalities of tea prepared by various thermal processing methods from Ixeris dentata root in order to elevate utilization of Ixeris dentata root as food. These methods included natural drying (ND), hot-air drying (HAD, 75℃), hot-air drying after steam (HADS, steaming at 95℃ for 30 min) and roasting after hot-air drying (RHAD, roasting at 140℃ for 2~3 min). Physiological functionalities of tea prepared by ND, HAD, HADS, and RHAD were measured as hot-water extracts and each tea was extracted by 100 ㎖ water on 1 g dried Ixeris dentata root powder. Electron donating abilities were the highest in HADS treatment at 98%. SOD-like activities were the highest in RHAD at 35.61%. Both ACE and tyrosinase inhibitory activities were the highest on ND treatment at 52.34% and 44.60%, respectively. Nitrite scavenging abilities of all treatment were the highest at pH 1.2 and the highest activity among them was on RHAD treatment at 50.5%.

      • KCI등재

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