• Journal of Crop Science and Biotechnology
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• 제11권1호
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• pp.45-50
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• 2008

Lignans and neolignans are optically active plant secondary metabolites. Research on biosynthesis of lignans has already been advanced especially for the formation of (+) pinoresinol but information on the biosynthesis of 8-O-4'- neolignans is still limited. Moreover, the chemical structure(position of substituents on aromatic rings) and stereochemistry of 8-O-4' neolignans is not clear. Katayama and Kado discovered that incubation of cell-free extracts from E. ulmoides with coniferyl alcohol in the presence of hydrogen peroxide gave (+)-erythro- and (-)-threo- guaiacylglycerol 8-O-4'-(coniferyl alcohol) ether (GGCE)(diastereomeric ratio, 3:2) which is the first report on enzymatic formation of optically active -8-O-4' neolignans from an achiral monolignol. In this aspect, enzymatic formation of guaiacyl 8-O-4' neolignan is noteworthy to clarify its stereochemistry from incubation of coniferyl alcohol with enzyme prepared from Eucommia ulmoides. In this experiment, soluble and insoluble enzymes prepared from E. ulmoides were incubated with 30 mM coniferyl alcohol(CA) for 60 min. The enzyme catalyzed GGCE, dehydrodiconiferyl alcohol(DHCA), and pinoresinol identified by reversed phase HPLC. Consequently, diastereomeric compositions of GGCE were determined as erythro and threo isomer. Enantiomeric composition was determined by the chiral column HPLC. Both enzyme preparations enantioselectively formed (-)-erythro, (+)-erythro and (+)-threo, (-)-threo-GGCEs respectively.

• 한국수산과학회지
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• 제42권6호
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• pp.537-544
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• 2009

This study was conducted to improve yield and functional properties of autoclave-treated salmon frame extracts (SFETA) using commercial enzymes (Alcalase 2.4 L FG, Flavourzyme 500 MG, Neutrase 0.8 L and Protamex 1.5 MG). Yield and angiotensin I converting enzyme (ACE) inhibitory activity of all enzymatic hydrolysates improved compared to those of control (undigested extracts), which were the highest in hydrolysates incubated with Protamex 1.5 MG for 4 hrs (P4-treated hydrolysates) and 2 hrs (P2-treated hydrolysates), respectively. However, antioxidant activities of all enzymatic hydrolysates showed less than 29%. According to the trichloroacetic acid soluble-N, volatile component intensity and sensory evaluation, when compared to control, taste of P4-treated hydrolysates improved, while its fish odor strongly smelt. Therefore, for efficient use of P4-hydrolysates, the fish odor should be improved by Maillard reaction of extracts or pre-treatment of salmon frame.

• 한국수산과학회지
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• 제27권2호
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• pp.133-139
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• 1994

식품의 가공 저장 및 조리 중에 일어나는 식품성분간의 반응 중 대표적인 반응으로 알려져 있는 Maillard 반응생성물의 돌연변이원성에 대한 그 억제 기구를 밝히기 위하여 glucose-arginine계 및 glucose-lysine${\cdot}$HCl계 Maillard 반응 생성물에 일상 생활에서 많이 섭취하고 있는 해조류 및 야채류의 수용성 추출물을 첨가하여 그 억제효과를 조사하였고, 아울러 이들 수용성추출물의 가열($100^{\circ}C$, 10분)에 따른 억제효과의 영향에 대해 조사 검토하였다. 그 결과, 실험에 사용한 해조류 및 야채류 수용성 추출물 전부가 돌연변이원성 억제효과를 나타내었고, 특히 양배추, 파, 산초, 및 청각에 있어서 억제효과가 높게 나타났다. 가열한 경우, 야채류의 돌연변이원성 억제효과는 감소하였으나 해조류는 크게 영향을 받지 않았다. 이 결과로 미루어 보아 당-아미노산계 Maillard 반응생성물의 돌연변이원성 억제에 해조류 및 야채류 추출물에 존재하는 환원성 물질, 고분자 물질 catalase 및 peroxidase와 같은 효소 등이 일조를 하는 것으로 나타났다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.133-148
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• 1987

Gibberellin(GA) 3-$\beta$ hydroxylation is very important for the shoot elogation in the higher plants, since only 3$\beta$-hydryoxylated GAs promote shoot elogation in several plants. Fluctuation of 3$\beta$-hydryoxylase activity was examined during seed maturation using two cultivars of , P. vulgaris, Kentucky Wonder (normal) and Masterpiece (dwarf). Very immature seeds of both cultivars contain high level of 3$\beta$-hydroxylase activity (per mg protein). Both cultivars showed maximum of enzyme activity (per seed) in the middle of their maturation process. Gibberellin 3$\beta$-hydroxylase catalyzing the hydroxylation of GA20 to GA1 was purified 313-fold from very early immature seeds of P. vulgaris. Crude soluble enzyme extracts were purified by 15% methanol precipitation, hydrophobic interaction chromatogrphy, DEAE ion exchange column chromatography and gel filtration HPLC. The 3$\beta$-hydroxylase activity was unstable and lost much of its activity duting the purification. The molecular weight of purified enzyme was extimated to be 42, 000 by gel filtration HPLC and SDS-PAGE. The enzyme exhibited maximum activity at pH 7.7. The Km values for [2.3-3H] GA20 and [2.3-3H]GA9 were 0.29 $\mu$M and 0.33 $\mu$M, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the Km value for 2-oxoglutarate was 250 $\mu$M using 3H GA20 as a substrate. Fe2+ and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Hg2+. Effects of several GAs and GA anaogues on the putrified 3$\beta$-hydroxylase were examined using [3H]GA9 and GA20 as a substrates. Among them, GA5, GA9, GA15, GA20 and GA44 inhibited the enzyme activity. [13C, 3H] GA20 was converted by the partially purified enzyme preparation to [13C, 3H]GA1, GA5 and GA6, which were identified by GC-MS, GA9 was converted only GA4, GA15 and GA44 were converted to GA37 and GA38, respectively. GA5 was epoxidized to GA6 by the preparation. This suggests that 3$\beta$-hydroxylation of GA20 and epoxidation of GA5 are catalyzed by the same enzyme in P, vulgaris.

• Journal of the Korean Wood Science and Technology
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• 제32권1호
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• pp.17-27
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• 2004

식용 임산자원인 산채류를 기능성 식품으로 개발하고자 산마늘을 비롯한 총 13종의 에탄올 조추출물에 대한 콜레스테롤 저하활성을 검정하였다. 즉, 콜레스테롤 생합성 과정 초기에 관여하는 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMG-CoA reductase)와 후기에 관여하는 squalene synthase의 효소 활성을 조사한 결과 산마늘 잎 에탄올 추출물이 두 효소의 활성을 공히 70% 이상 저해하여 활성이 가장 우수한 것으로 나타났다. 그리고 이러한 콜레스테롤 저하활성과 관련이 있는 물질을 탐색하고자 산마늘로부터 물질 단리를 시도하여 디클로로메탄 가용부로부터 kaempferol과 quercetin을 단리하였다. 또한 유전자 레벨에서의 콜레스테롤 저하 활성을 조사하기 위해 단리물질과 분획물을 C100세포(햄스터 유래 HMG-CoA reductase 고발현 세포주)에 각각 5 ㎍/㎖과 10 ㎍/㎖로 24시간 처리하여 HMG-CoA reductase와 squalene synthase의 mRNA 발현 정도를 조사하였다. 그 결과 10 ㎍/㎖로 kaempferol과 quercetin을 처리한 경우 두 효소의 mRNA가 전혀 발현하지 않는 것으로 나타나 유전자 레벨에서의 콜레스테롤 생합성 저해 효과를 확인할 수 있었다. 이상의 결과 산마늘 잎 에탄올 추출물은 콜레스테롤 생합성에 관여하는 HMG-CoA reductase와 squalene synthase의 활성을 저해하며 이러한 활성 저해 효과는 kaempferol과 quercetin에 기인하는 것으로 사료되었다. 특히 kaempferol과 quercetin은 여러 식물의 성분으로서 이미 알려진 화합물이지만 콜레스테롤 저하활성이 있는 것으로 밝혀진 것은 이번이 처음으로 금후 이들 물질과 이들 물질을 함유하고 있는 식물 활용에 필요한 자료를 제공하였다고 사료된다.

• Journal of Nutrition and Health
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• 제34권3호
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• pp.285-298
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• 2001

The effects of dried leaf powders and water and ethanol extracts of persimmon and green tea on lipid metabolism, lipid peroxidation and antioxidative enzyme activity were investigated in 12-month-old rats. Forty-nine male Sprague-Dawley rats weighing 520$\pm$19g were blocked into seven groups according to body weight. Rats were raised for four weeks with control(no tea leaf powder or extracts) and experimental diets containing either 5%(w/w) dried leaf powders of persimmon(Diospyros kaki Thunb) or green tea(Camellia sinensis O. Ktze), or water or ethanol extract from equal amounts of each dried tea powder. Food intakes of all tea diet groups were higher than that of control. Weight gains and food efficiency ratios of all tea diet groups were not significantly different from those of control. All tea diets decreased plasma triglyceride level, especially, green tea powder and persimmon ethanol diets were more effective than other diet. All the tea diet groups showed decrease in liver triglyceride level, and persimmon powder and ethanol extract increased fecal triglyceride excretion. Plasma cholesterol levels of all the tea diet groups were not significantly different from the control, but control. Fecal cholesterliver cholesterol concegroups were significantlntrations of all tea y lower than that of ol excretions of persimmon powder, green tea ethanol extract, persommon ethanol extract and green tea ethanol extract groups were significantly higher than that of control. Plasma and liver thiobarbituric acid reactive substance(TBARS) concentrations of all the tea diet groups were lower than that of control. Especially, plasma TBARS concentrations of green tea powder and persimmon ethanol extract groups were sinificantly low. Red blood cell(RBC) superoxide dismutase(SOD) activities of persimmon ethanol extract and green tea water extract groups were increased, and RBC catalase activities of all experimental groups were not significantly different. RBC glutathione peroxidase(GSH-px) activities of persimmon ethanol extract, persimmon water extract and green tea powder groups were increased. Liver SOD activities of all the tea diet groups except green tea ethanol extract group were higher than that of control. Liver catalase activities of all experimental groups were not significantly different, and liver GSH-px activity of green tea powder group was significantly higher than that of control. In conclusion, dried leaf powders, and water and ethanol extracts of persimmon and green tea were effective in lowering lipid level, inhibiting lipid peroxidation and increasing antioxdative enzyme activities in 12-month-old rat. Green tea leaf powder with high contents of flavonoids and water soluble dietar fiber was most effective in lowering plasma triglyceride, cholesterol and TBARS level. (Korean J Nutrition 34(3) : 285~298, 2001)

• Journal of Crop Science and Biotechnology
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• 제10권1호
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• pp.19-26
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• 2007

Glutathione S-transferases(GSTs, EC 2.5.1.18), glyoxalase-I(EC 4.4.1.5) and alliin lyase(alliinase, EC 4.4.1.4) are important enzyme systems in plant bodies. The first two are mainly detoxifying enzymes that utilize glutathione(GSH) in the defense mechanism, and the last one is mainly involved in secondary metabolism and relevant to sulfur compounds derived from GSH. The activities of the three enzymes have been investigated in soluble extracts of vegetable crops, including pumpkin, cabbage, broccoli, radish, carrot, potato, sweet potato, mungbean, and onion. GST activities were detected in all of the vegetables, and the extract of onion bulb exhibited the highest specific activity(648 nmol/min/mgP). The putative GSTs of most of the vegetables were found to be induced by ethanol. The activities of GSTs in onion bulb were found to be markedly inhibited by S-hexyl glutathione and were also inhibited by S-butyl glutathione and S-propyl glutathione. The anti-CmGSTF1 antiserum recognized a thick band for putative onion GST. The estimated glyoxalase-I activity level was also high in onion bulb(4540 nmol/min/mgP), indicating that the thick band detected by Western blot analysis might result from partial recognition of glyoxalase-I by the antiserum. The specific activities for glyoxalase-I were moderate in radish and carrot, and the extracts of other vegetables had rather low levels of activities. The extract of onion also showed the highest specific activity level for alliinase(2069nmol pyruvate/mgP). The extracts of other vegetables also had alliinase activities, although the estimated values were much lower than that of onion.

• 한국식품저장유통학회지
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• 제20권6호
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• pp.784-790
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• 2013

가압가열추출(AE), 열수추출(HWE) 및 효소처리추출(EE) 방법으로 추출한 포도 추출액의 품질을 조사하였다. 추출수율은 가압추출액이 79.8%, 열수추출액이 82,3% 및 효소추출액이 92.6%로 효소 추출이 가장 높은 수율을 나타내었다. pH는 추출 방법에 따른 유의적인 차이는 없었으며, 산도와 당도는 효소추출에서 유의적으로 높은 함량을 보였다. 색상에 있어서는 $a^*$과 $b^*$값은 효소 추출이 5.24 및 5.09로서 열수추출(3.79, 3.83)과 가압가열추출(1.83, 3.16)보다 높은 값을 보였으며 $L^*$값은 차이를 보이지 않는 것으로 나타났다. 점도는 효소추출이 42.1 cP 이었으며 가압가열추출과 열수추출보다 높았다. 유리당은 포도당과 과당의 함량이 높으며, 가압추출과 열수추출보다 효소추출이 높은 함량을 나타내었다. 유기산은 사과산의 함량이 가장 많았으며 효소 추출이 가장 높았다. 총 폴리페놀의 함량은 효소추출이 55.7 mg% 수준으로 가압추출 31.3 mg%와 열수추출 39.5 mg%에 비하여 높았으며, 특히 안토시아닌 함량은 효소추출이 22.1 mg%로 가압추출과 열수추출보다 2~4배 높은 함량을 보였다. 항산화 활성 측정 결과 DPPH 라디칼 소거 활성과 환원력에 있어서 효소추출이 가압추출과 열수추출에 비하여 높은 활성도를 나타내었다. 결과적으로 포도 추출에 있어 가압과 가열에 의한 방법보다 효소 처리에 의한 추출이 맛과 색소 성분의 추출을 용이하게 하며 그에 따른 항산화 활성도 높이는 것으로 나타났다.

• Fisheries and Aquatic Sciences
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• 제26권3호
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• pp.204-215
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• 2023

The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10^-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.184-184
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• 1998

Aldose reductase(AR), a rate-limiting enzyme in the polyol pathway, has been demonstrated to cause the intracellular accumulation of sorbitol or galactitol and hence to play key roles not only in the cataract formation in the lens but also in the pathogenesis of diabetic complications such as neuropathy, retinopathy and nephropathy, etc. In a series of investigations to evaluate potential AR inhibitors from medicinal plants, we have shown that some hot water extracts exhibited a significant inhibition of a significant inhibition of bovine lens AR in vitro. Among active plants, the roots of Angelica dahuria (Umbelliferae) were shown to have relatively potent AR inhibitory activity. Systematic fractionation of the ether soluble fraction monitored by bioassay led to isolation of two furanocoumarins, byakangelicin(I) and ter-O-methyl byakangelicin( II), were identified as potential AR inhibitors, their $IC_{50}$ values being 6.2 M and 2.8 M, respectively.