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.

성숙과정 중 녹색밀의 이화학적 특성변화

김민철(Min?Chul Kim),이가순(Ka?Soon Lee),이병진(Byung?Jin Lee),권병구(Byung?Gu Kwon),주정일(Jung?Il Ju),구자형(Ja?Hyeong Gu),오만진(Man?Jin Oh) 한국식품영양과학회 2007 한국식품영양과학회지 Vol.36 No.10

미성숙 상태의 녹색밀을 식품가공소재로 이용하기 위한 영양적 기초자료를 얻고자 밀이 성숙과정 중에 일반성분, 유리당, 유리아미노산, β-glucan, 무기물 및 색도를 측정하였다. 공시품종인 금강밀, 그루밀, 청계밀의 수분, 조단백, 조지방, 회분은 완숙에 가까워질수록 감소하는 경향을 보여 주었고 조섬유의 함량은 금강밀은 3.0%에서 3.26%로, 그루밀은 3.35%에서 4.59%로, 청계밀은 4.25%에서 6.11%로 각각 증가하였다. 출수 25일 경에 밀의 유리당 함량은 maltose, maltotriose가 높았고 금강밀, 그루밀, 청계밀 모두 완숙에 가까워질수록 감소하였다. 유리 아미노산 함량은 금강밀, 그루밀, 청계밀 모두 출수 후 25일경에는 alanine, GABA, glycine 순으로 많았고 완숙밀에 가까워질수록 각각의 유리 아미노산은 감소하는 경향을 보였으나 glutamic acid만이 증가하는 경향을 보여 성숙한 밀에서는 alanine, glutamic acid, GABA가 많았다. 셋 품종 중 그루밀이 유리아미노산 함량이 비교적 높았다. 모든 공시재료에서 완숙에 가까워질수록 β-glucan함량은 감소하는 경향을 보여 주었다. 금강밀의 출수 후 26일 경 β-glucan 함량은 1.5%에서 출수 후 43일에는 0.28%로 셋 품종 중 가장 크게 감소하였으며 모든 품종에서 같은 경향이었다. 출수 후 완숙에 가까워질수록 공시밀의 무기물 함량은 전반적으로 증가하였고 특히 K, Mg의 증가하는 경향은 뚜렷하였다. 공시품종 모두에서 완숙에 가까워질수록 L값은 감소하였으며 a값과 b값은 증가하였으며 출수 후 33일 경까지 녹색을 유지하고 증미성 물질이 다량 존재하여 가공식품의 소재를 이용하는 것이 바람직하다. In order to evaluate the utilization of immature green wheat kernels as food processing material, these experiments were carried out to analyze general composition, free sugars, free amino acids, mineral content and color of 3 wheat varieties according to their heading dates. Contents such as moisture, crude protein, crude lipid and ash were gradually decreased according to the days after heading of 3 wheat varieties, while crude fiber and starch were gradually increased. Maltose and maltotriose contents in immature green wheat kernels were high but its rapidlly decreased by closing to ripeness. Contents of free amino acids showed a tendency to decrease, among which alanine content was the highest around 25 days after heading, and then glutamic acid, GABA, glycine in order. Contents of total free amino acid were decreased by closing to ripeness. Among 3 wheat varieties, its contents of Guru wheat was the highest. As wheat matured, β-glucan contents were decreased from 1.5% to 0.28% on 43 days after heading. Mineral contents of 3 wheat varieties were generally increased by closing to ripeness after coming into ears, and K, Mg were especially prominent. As wheat matured, lightness was decreased, while redness and yellowness were increased. Considering chemical composition and color, the immature green wheat were produced through blanching the spikes harvested before the yellow ripe stage. The harvested green wheat cereals is able to eat raw wheat or cook it as food processing material.

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.