• Title/Summary/Keyword: 폴리페놀 산화효소

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Influence of Ph and Temperature on Polyphenol Oxidase in the Leaves of Perilla frutescens var. japonica (들깨잎 폴리페놀 산화효소의 pH 및 온도에 의한 영향)

  • Kim, Yoo-Kyung;Kim, An-Keun
    • YAKHAK HOEJI
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    • v.48 no.6
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    • pp.384-390
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    • 2004
  • Polyphenol oxidase-catalyzed oxidation of substrates (t-butylcatechol, 4-methylcatechol, chlorogenic acid, caffeic acid and pyrocatechol) were performed in the Ph range 4~8. Co ncentrations of substrate's major oxidation products were monitored by high performance liquid chromatograph. The nature and amounts of products formed were highly pH dependent. They also were ifluenced by kinds of substrates. Major oxidation product of 4-methylcatechol appeared the maxium value at pH 5, them of chlorogenic acid, caffeic acid and pyrocatechol at pH 6.0 and that of t-butylcatechol at pH 5~7. Time-dependent PPO activity was determined at $4^{\circ}C\;and\;30^{\circ}C$. PPO extracted by phosphate buffer containing triton X-114 (t-PPO) was more stable than PPO by phosphate buffer (b-PPO). The result of electrophoresis, at first PPO was showed only a band at 48 kd. After 1~3 days a partial degrade band was appeared in b-PPO and three partial degrade bands in t-PPO. No activity band was appeared in PPOs at $30^{\circ}C$ and b-PPO at $4^{\circ}C$ after 4 days. And a band (37 kDa) in t-PPO was remained finally and disappered. PPO from Perillae leaves has two activity bands at 48 and 37 kDa in previous paper. It was supposed that PPO in the leaves of Perilla frutescens was a protein having one molecular weight as 48 kDa. And 37 kDa protein, relatively proteolysis-resistant, was a proteolyzed form of a major form.

Antioxidative Activity of Mushroom Water Extracts Fermented by Lactic Acid Bacteria (유산균 발효에 의한 버섯 추출물의 항산화 활성)

  • Yang, Hee Sun;Choi, Yu Jin;Oh, Hyun Hee;Moon, Joon Seong;Jung, Hoo Kil;Kim, Kyung Je;Choi, Bong Suk;Lee, Jung Won;Huh, Chang Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.80-85
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    • 2014
  • This study was focused on the development of fermented mushroom water extracts with antioxidative activities. Mushroom water extracts were fermented with Bifidobacterium bifidum, Lactobacillus plantarum, Lactobacillus acidophilus, Leuconostoc lactis, Streptococcus thermophilus and Lactobacillus sakei subsp. LI033 was isolated from kimchi. Fermented mushroom water extracts increased DPPH and ABTS radical scavenging activities in a dose-dependent manner. However, radical scavenging activity of fermented Phellinus linteus and Ganoderma lucidum water extracts was decreased compared to non-fermented mushroom water extracts. Antioxidative activity of fermented mushroom water extracts was also confirmed by xanthin oxidase (XO) inhibition and superoxide dismutase (SOD) activities at the same concentration. As the fermentation progressed, fermented mushroom water extracts increased XO inhibition activity and SOD activity. In conclusion, fermented mushroom water extracts were tentatively identified to enhance enzyme activity.

Protective Effects of Perilla frutescens Britt var. japonica Extracts from Oxidative Stress in Human HaCaT Keratinocytes (HaCaT 피부각질세포에서 들깻잎 추출물의 산화적 스트레스에 대한 항산화 효과)

  • Ji, Na;Song, Jia-Le;Kil, Jeung-Ha;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.161-167
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    • 2013
  • The aim of this study was to investigate the protective effects of methanolic extract from perilla (Perilla frutescens Britt var. japonica) leaves (PLME) on oxidative injury from hydrogen peroxide ($H_2O_2$) in human HaCaT keratinoctyes. Cells were co-incubated with various concentrations (0~200 ${\mu}g/mL$) of PLME for 24 hr, and then exposed to $H_2O_2$ (500 ${\mu}M$) for 4 hr. $H_2O_2$ significantly decreased cell viability (p<0.05). However, PLME provided protection from $H_2O_2$-induced HaCaT cell oxidation in a dose-dependent manner. To further investigate the protective effects of PLME on $H_2O_2$-induced oxidative stress in HaCaT cells, the cellular levels of lipid peroxidation, and antioxidant enzymes (including superoxide dismutase (SOD), glutathione peroxidase (GSH-px) and catalase (CAT)) were measured. PLME decreased cellular levels of lipid peroxidation, and also increased the activities of antioxidant enzymes. In addition, the antioxidant activities of PLME were also determined by DPPH and hydroxyl (${\cdot}OH$) radical scavenging assay, and major antioxidant compounds of PLME were measured by colorimetric methods. DPPH and ${\cdot}OH$ radical scavenging activities of PLME increased in a dose dependent manner and was similar to the DPPH scavenging activity of ascorbic acid at 50 ${\mu}g/mL$; however PLME activities were stronger than ascorbic acid (50 ${\mu}g/mL$) in the ${\cdot}OH$ scavenging assay. The amounts of antioxidant compounds, including total polyphenolics, total flavonoids, and total ascorbic acid from PLME were $52.2{\pm}1.1$ mg gallic acid (GAE)/g, $33.7{\pm}4.7$ mg rutin (RUE)/g, and $17.0{\pm}0.5$ mg ascorbic acid (AA)/g, respectively. These results suggest that PLME has a strong free radical-scavenging activity and a protective effect against $H_2O_2$-induced oxidative stress in the keratinocytes.

Inhibition of Enzymatic Browning of Crown Daisy by Maillard Reaction Products (마이얄반응생성물(Maillard reaction product)에 의한 쑥갓의 효소적갈면 억제)

  • Kim, Ji-Hae;Song, Hyeon-Seung;Park, Inshik
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1451-1455
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    • 2012
  • The study was conducted to investigate the effect of Maillard reaction products (MRPs) on enzymatic browning of crown daisy (Chrysanthmum coronarium var. spatiosum). The MRPs prepared by heating various amino acid and sugar at $90^{\circ}C$ caused a strong inhibitory effect on crown daisy polyphenol oxidase (PPO, ${\sigma}$-diphenol oxygen oxidoreductase, EC 1.10.3.1). As the reaction time of the solution containing glycine and glucose increased at $90^{\circ}C$, the production of MRPs was increased, whereas the amounts of glycine and glucose were decreased. Accordingly, the inhibitory effect of crown daisy PPO activity by MRPs was increased as the amounts of synthesized MRPs were increased. The MRPs synthesized from the various amino acids and sugars significantly reduced the PPO activity, particularly MRPs prepared by glutamine and xylose. The Michealis-Menten constant value ($K_m$) of crown daisy PPO with catechol as a substrate was 22.0 mM, and MRPs were a noncompetitive inhibitor against crown daisy PPO.

Characterization of Laccase Purified from Korean Trametes hirsuta S1 (한국산 흰구름버섯(Trametes hirsuta S1)으로부터 정제된 Laccase의 특성)

  • Lim, Hyung-Seon;Yoon, Kyung-Ha
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.112-118
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    • 2004
  • Laccase produced by Trametes hirsuta S1 isolated from Korea was partially purified and characterized using ultrafiltration, anion exchange chromatography and affinity chromatography. The laccase was produced as the predominant extracellular enzyme during primary metabolism. Neither lignin peroxidase nor veratryl alcohol oxidase (VAO) were detected in the culture fluid. Addition of 2,5-xylidine enhanced 4-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 12%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to $51.2\;{\mu}M\;and\;56.8\;{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $50^{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 20 min at $70^{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the T. hirsuta S1 laccase showed 100% of homology to those of laccase from C. hirsutus.

Ethyl acetate fraction from Pteridium aquilinum ameliorates cognitive impairment in high-fat diet-induced diabetic mice (고지방 식이로 유도된 실험동물의 당뇨성 인지기능 장애에 대한 고사리 아세트산에틸 분획물의 개선효과)

  • Kwon, Bong Seok;Guo, Tian Jiao;Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Park, Sang Hyun;Kang, Jeong Eun;Lee, Chang Jun;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.649-658
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    • 2017
  • The potential of the ethyl acetate fraction from Pteridium aquilinum (EFPA) to improve the cognitive function in high-fat diet (HFD)-induced diabetic mice was investigated. EFPA-treatment resulted in a significant improvement in the spatial, learning, and memory abilities compared to the HFD group in behavioral tests, including the Y-maze, passive avoidance, and Morris water maze. The diabetic symptoms of the EFPA-treated groups, such as fasting glucose and glucose tolerance, were alleviated. The administration of EFPA reduced the acetylcholinesterase (AChE) activity and malondialdehyde (MDA) content in mice brains, but increased the acetylcholine (ACh) and superoxide dismutase (SOD) levels. Finally, kaempferol-3-o-glucoside, a major physiological component of EFPA, was identified by using high-performance liquid chromatography coupled with a hybrid triple quadrupole-linear ion trap mass spectrometer (QTRAP LC-MS/MS).

Protective effect of ethyl acetate fraction from Actinidia arguta sprout against high glucose-induced in vitro neurotoxicity (포도당으로 유도된 in vitro 뇌신경세포 독성에 대한 다래 순 아세트산에틸 분획물의 보호 효과)

  • Yoo, Seul Ki;Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Park, Su Bin;Han, Hye Ju;Kim, Chul-Wo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.517-527
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    • 2018
  • The current study investigated in vitro anti-diabetic and neuroprotective effects of the ethyl acetate fraction in Actinidia arguta sprouts (EFAS), on $H_2O_2$ and high glucose-induced cytotoxicity in human neuroblastoma MC-IXC cells. EFAS had high total phenolic and total flavonoid contents. An assessment of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity of EFAS, as well as its potential for inhibiting malondialdehyde production, indicated that EFAS may possess significant antioxidant properties. EFAS exerted inhibitory effects on ${\alpha}-glucosidase$ via glycemic regulation which forms advanced glycation end products. In addition, EFAS exhibited significant acetylcholinesterase inhibitory effects. Moreover, EFAS displayed protective effects against $H_2O_2$ and high glucose-induced cell death, and inhibited the generation of reactive oxygen species in MC-IXC cells. Finally, the main physiological compound of EFAS was identified via high performance liquid chromatography as a rutin.

Effects of (-)-Epigallocatechin-3-gallate on the Release of Pancreatic Enzymes and Expression of Regenerating Genes in Ethanol-injured Murine Pancreatic Primary Acinar Cells (에탄올에 의하여 유도된 마우스 췌장 선포세포의 염증성 손상에서 췌장분비 효소의 활성 및 세포 재생관련 유전자들의 발현에 미치는 EGCG의 영향)

  • Kim, Sung Ok;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1404-1408
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    • 2013
  • (-)-Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, has been shown to have strong antibacterial, antiviral, antioxidant, anti-inflammatory, and chemopreventive effects. However it is unknown whether EGCG can recover alcohol-associated pancreatitis. The aim of this study was to investigate the effects of EGCG on pancreatic enzyme activities and the expressions of pancreatic regenerating related markers, such as adenosine monophosphate-activated protein kinase (AMPK), raf-1 kinase inhibitor protein (RKIP), and Regenerating gene 1 (Reg1), in mice pancreatic primary acinar cells. Our results revealed that activities of ${\alpha}$-amylase and chymotrypsin were significantly increased in the cells treated with ethanol compared to the untreated control cells; however, the increased activities of both enzymes were markedly reduced by pretreatment with EGCG. Phosphorylation of AMPK and total expression of RKIP were decreased in the ethanol-treated primary acinar cells; however, these were both significantly increased in the EGCG-pretreated cells. In addition, when EGCG was treated, expression of Reg1 was markedly increased compared with that of the control or the ethanol-treated primary acinar cells, demonstrating that EGCG can modulate pancreatic regenerating related genes. Therefore, our findings suggest that EGCG may have therapeutic utility in the prevention or treatment of alcohol-associated pancreatitis.

Antioxidant activity and neuroprotective effects of ethanol extracts from the core of Diospyros kaki (감 심지 에탄올 추출물의 항산화 활성 및 신경세포 보호 효과)

  • Byun, Eui-Baek;Kim, Min-Jin;Kim, Soon-Jung;Oh, Nam-Soon;Park, Sang-Hyun;Kim, Woo Sik;Song, Ha-Yeon;Han, JeongMoo;Kim, Kwangwook;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.60-66
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    • 2020
  • This study examined the antioxidant activity and neuroprotective effects of ethanol extracts obtained from Diospyros kaki core (DCE). The total polyphenol and flavonoid contents in DCE was 786.47±15.27 and 31.14±0.82 mg/g, respectively. In addition, DCE exhibited a dose-dependent induction of radical scavenging activity, determined by 1,1-diphenyl-picrylhydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonicacid) (ABTS), ferric reducing antioxidant power (FRAP), and reducing power assays. The viability of HT22 hippocampal cells was examined to investigate the neuroprotective effect of DCE. DCE treatment did not induce cytotoxicity at concentrations below 1,000 ㎍/mL. Additionally, DCE treatment in the background of H2O2 induce oxidative stress revealed a significant increase in the survival rat, indicated by increased SOD activity and decreased levels of MDA, a lipid peroxidation product. Therefore, the results suggest that DCE can be used as a source of natural antioxidants source and a therapeutic agent for the treatment of brain disorders induced by oxidative stress and neuronal damage.

Effects of water extract of Paeoniae Radix Alba on a thioacetamide induced acute liver injury rat model (Thioacetamide 유발 급성 간손상 동물모델에 백작약 열수 추출물이 미치는 효능)

  • Lee, Se Hui;Shin, Mi-Rae;Lee, Ji Hye;Roh, Seong-Soo
    • Journal of Nutrition and Health
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    • v.54 no.2
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    • pp.224-237
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    • 2021
  • Purpose: Paeonia Radix Alba is a traditional herbal medicine used to treat the liver and the spleen. Many studies have reported that Paeonia Radix Alba extract (PR) affects liver injury, but there has been no study on liver injuries induced by thioacetamide (TAA). Therefore, we aimed at evaluating the effect of PR on a TAA-induced acute liver injury (ALI) model. Methods: The antioxidant activity of PR was assayed by the content of total polyphenol, total flavonoid, 1,1-diphenyl-2'-picrylhydrazyl (DPPH), and 2,2'-azino-bis (3-ethylbenzo-thiazoline-6-sulfonicacid) (ABTS) radical scavenging activities in vitro test. ALI was induced via-intraperitoneal injection of TAA (200 mg/kg body weight) for three consecutive days. Also, silymarin (100 mg/kg body weight) and PR (100 or 200 mg/kg body weight) were administered at 1 hours 30 minutes prior to TAA treatment. The levels of ammonia, glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) were analyzed using an assay kit. The expressions of antioxidant proteins including Nrf2, Keap1, HO-1, SOD, catalase, and GPx-1/2 and oxidative stress-related proteins including NOX2, p47phox, and p22phox were evaluated by the western blot analysis. Results: PR showed excellent antioxidant activity in vitro. TAA administration increased the levels of ammonia, GOT, and GPT in the ALI control group compared to the normal group, whereas it was significantly reduced by PR pretreatment. Moreover, NADPH oxidase protein expressions were upregulated after TAA treatment, while the elevated expressions were inhibited by PR pretreatment. The expressions of antioxidant protein were downregulated in the ALI control group, whereas Nrf2 activation in the PR group was accompanied by increased levels of antioxidant enzymes. Conclusion: PR administration increased the antioxidant enzymes via activation of the Keap1/Nrf2 pathway and inhibited the protein levels of NADPH oxidase factors. Taken together, these results showed that PR treatment may be considered to ameliorate acute liver injury induced by TAA.