• Title/Summary/Keyword: enzymatic extracts

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Effect of Medicinal Herb Prepared through Traditional Antidiabetic Prescription on α-Glucosidase Activity and Evaluation Method for Anti-Melanogenesis Agents Using α-Glucosidase Activity (당뇨 처방에 근거한 생약재의 α-Glucosidase 활성 저해 효과 및 이를 활용한 미백 소재 평가법)

  • Kim, Mi Jin;Im, Kyung Ran;Yoon, Kyung-Sup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.7
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    • pp.993-999
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    • 2015
  • For the purpose of investigating the in vitro antidiabetic activity of a medicinal herb and herb mixture extracts prepared through traditional antidiabetic prescription, this study examined ${\alpha}$-glucosidase inhibitory activity. Tyrosinase, a type I membrane glycoprotein, is synthesized and glycosylated in the endoplasmic reticulum (ER) and Golgi. The enzyme is subsequently transported to melanosomes, where it participates in melanogenesis. Previous studies showed that disruption of early ER N-glycan processing by an ${\alpha}$-glucosidase inhibitor suppresses tyrosinase enzymatic activity and melanogenesis. According to the results, most oriental medicinal herbal extracts were stronger than acarbose and N-butyldeoxynojirimycin, known as an ${\alpha}$-glucosidase inhibitor. Interestingly, ethyl acetate layer of enzyme hydrolyzed Cheongsimyeonjaeum had an inhibitory effect on melanin synthesis in B16F1 cells, although it did not inhibit tyrosinase activity directly. Together, ${\alpha}$-glucosidase inhibition activity could be used to evaluate anti-melanogenesis, although cross-checking with melanin inhibitory assay is recommended.

Optimal Extraction and Characteristics of Lignocellulytic Enzymes from Various Spent Mushroom Composts (다양한 버섯 수확 후 배지로부터 목질섬유소 분해효소의 최적 추출 및 특성)

  • Lim, Sun Hwa;Lee, Yun Hae;Kang, Hee Wan
    • The Korean Journal of Mycology
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    • v.41 no.3
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    • pp.160-166
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    • 2013
  • Recovery of ${\alpha}$-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) from spent mushroom composts (SMCs) of Pleurotus cornucopiae, Pleurotus ostreatus, Pleurotus eryngii, Hericium erinaceum, Lyophyllum ulmarium, Agrocybe cylindracea, Lentinus lepideus, and Flammulina velvtipes were investigated using different extraction buffers. The maximum recovery of the enzymes was mostly detected in SMC extracts with tap water and 0.25% Triton X-100 by shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. The xylanase (152 U/g) and laccase (8.1 U/g) activities were the highest in SMC extracts from F. velvtipes and P. eryngii. In addition, high enzymatic activities of ${\alpha}$-amylase (3.6 U/g) and cellulase (3.4 U/g) was detected in SMC extract of A. cylindracea. Futhermore, cellulase and laccase activities of SMCE from P. eryngii were compared to commercial enzymes.

Antioxidant Activity and Identification of Lunasin Peptide as an Anticancer Peptide on Growing Period and Parts in Pepper (생육시기 및 부위별 고추의 항산화력 및 항암 Lunasin peptide의 동정)

  • Kwon Ki Soo;Park Jae Ho;Kim Dae Seop;Jeong Jin Boo;Sim Young Eun;Kim Mi Suk;Lee Hee Kyung;Chung Gyu Young;Jeong Hyung Jin
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.528-535
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    • 2005
  • The non-enzymatic anti-oxidants and lunasin peptide from the extracts of the pepper were examined in order to utilize the discovery in natural products as cancer chemopreventive agents. The DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging activity on the fruit parts of the pepper was higher than that of the seed, but the difference was low. The Inhibition activity of xanthine/ xanthine oxidase in extracts of the seed was higher than that of the fruit and that of the seed on 20 days after flowering was the highest at the growing period. These were identified as fatty acids and phenolic compounds such as 1-eicosanol, palmitic acid, linoleic acid, linolenic acid and benzonitrile. The contents of fatty acids and phenolic compounds increased according to the time passing at the growing period. Peroxidase (POD) activity of the fruit at middle stage was high than that of other growing stages and that of the seed was the highest at later growing period. Though superoxide dismutase (SOD) activities in fruit were hish by passage of Slowing stage, the activity in seed was low. Lunasin was searched from seeds of the peppers by coomassie blue staining and western blot among them and we just found lunasin peptide from extracted protein of the pepper by western blot. In addition, we observed the contents of lunasin after flowering and confirmed to appear the lunasin at 35 days after flowering. We confirmed that lunasin is complex protein of maturing seeds. 100nM lunasin peptide in pepper showed inhibition effect on colony formation in $2\~12$ cells.

Change in Quality of Mixed Juice of Fruits and Vegetables by Aseptic Treatment and Packing with Nitrogen Gas during Storage (제균처리와 질소가스포장에 따른 혼합과채주스의 저장 중 품질 변화)

  • Kim, Su-Yeun;Yoon, Young-Bean;Choi, Eon-Ho
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1271-1277
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    • 2000
  • The extracts from tomatoes, apples, carrots, mallows, watercreses+pine needles, Angelica keiskei Koiz, jujubes and lemons were selected and mixed at the ratio of 3 : 3 : 3 : 1/2 : 1/2 : 1/2 : 1/2 : 1/5 on the basis of sensory evaluation. The mixed extracts were divided into three lots and treated as follows. The first lot was heated for 15 sec at $96^{\circ}C$ without centrifugation and filtration, the second filtered through a ultramembrane filter. For the third one, the ultrafiltrate combined with autoclaved retentates on the membrane filter and the previous centrifugal precipitation. The mixed juices were stored in the glass bottles with atmosphere or in film package with nitrogen gas. And then they were stored at $4^{\circ}$ and $20^{\circ}C$. During storage the treated juices showed $pH\;4.07{\sim}4.10$, titratable acidity $66.35{\sim}84.08$, soluble solid $7{\sim}9^{\circ}Brix$, reducing sugar $5.42{\sim}6.97%$, glucose $1.96{\sim}2.30%$ and fructose $3.46{\sim}4.14%$. The ultrafiltered juices showed yellow color, different from orange color of other treatment lots. Peroxidase activity and microbial population were inhibited by thermal treatment and ultrafiltration. The browning effects of the mixed juice of fruits and vegetables during storage were caused by ascorbic acid oxidation and non-enzymatic browning reaction, which could be reduced by change and packing with nitrogen gas instead of atmosphere inside bottles.

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The responsibility of C-terminal domain in the thermolabile haemolysin activity of Vibrio parahaemolyticus and inhibition treatments by Phellinus sp. extracts

  • Tran Thi Huyen;Ha Phuong Trang;Nguyen Thi-Ngan;Bui Dinh-Thanh;Le Pham Tan Quoc;Trinh Ngoc Nam
    • Fisheries and Aquatic Sciences
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    • v.26 no.3
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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.

Development and Characterization of High Quality Salted Mackerel Using Enzyme Hydrolysates of Ecklonia cava (감태 효소 가소분해물을 이용한 고품질 간고등어의 개발 및 특성)

  • Yoon, Min-Seok;Kim, Hyung-Jun;Park, Kwon-Hyun;Shin, Joon-Ho;Lee, Jeong-Suk;Jeon, You-Jin;Son, Hee-Jin;Heu, Min-Soo;Kim, Jin-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.6
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    • pp.545-554
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    • 2009
  • This study was conducted to develop and characterize of a high quality salted mackerel using enzymatic extracts of Ecklonia cava (EEC). In this study, potential antioxidative properties of EEC were evaluated by DPPH free radical scavenging activity, hydrogen peroxide scavenging activity, peroxide value, and fatty acid composition, and the antimicrobial properties were also measured by analysis for volatile basic nitrogen, pH, viable cells, Eschericia coli and biogenic amine. Compared to EEC-untreated salted mackerel, the salted mackerel with EEC was superior in antioxidative properties, while was negligible in the difference of antimicrobial properties. These results suggested that the high quality salted mackerel with antioxidative activity could be developed by treatment of EEC.

Pig Skin Gelatin Hydrolysates Attenuate Acetylcholine Esterase Activity and Scopolamine-induced Impairment of Memory and Learning Ability of Mice

  • Kim, Dongwook;Kim, Yuan H. Brad;Ham, Jun-Sang;Lee, Sung Ki;Jang, Aera
    • Food Science of Animal Resources
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    • v.40 no.2
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    • pp.183-196
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    • 2020
  • The protective effect of pig skin gelatin water extracts (PSW) and the low molecular weight hydrolysates of PSW generated via enzymatic hydrolysis with Flavourzyme® 1000L (LPSW) against scopolamine-induced impairment of cognitive function in mice was determined. Seventy male ICR mice weighing 20-25 g were randomly assigned to seven groups: Control (CON); scopolamine (SCO, 1 mg/kg B.W., intraperitoneally (i.p.); tetrahydroaminoacridine 10 [THA 10, tacrine; 10 mg/kg B.W. per oral (p.o.) with SCO (i.p.)]; PSW 10 (10 mg/kg B.W. (p.o.) with SCO (i.p.); PSW 40 (40 mg/kg B.W. (p.o.) with SCO (i.p.); LPSW 100 (100 mg/kg B.W. (p.o.) with SCO (i.p.); LPSW 400 (400 mg/kg B.W. (p.o.) with SCO (i.p.). All treatment groups, except CON, received scopolamine on the day of the experiment. The oxygen radical absorbance capacity of LPSW 400 at 1 mg/mL was 154.14 μM Trolox equivalent. Administration of PSW and LPSW for 15 weeks did not significantly affect on physical performance of mice. LPSW 400 significantly increased spontaneous alternation, reaching the level observed for THA and CON. The latency time of animals receiving LPSW 400 was higher than that of mice treated with SCO alone in the passive avoidance test, whereas it was shorter in the water maze test. LPSW 400 increased acetylcholine (ACh) content and decreased ACh esterase activity (p<0.05). LPSW 100 and LPSW 400 reduced monoamine oxidase-B activity. These results indicated that LPSW at 400 mg/kg B.W. is a potentially strong antioxidant and contains novel components for the functional food industry.

Glycosylation Enhances the Physicochemical Properties of Caffeic Acid Phenethyl Ester

  • Moon, Keum-Ok;Park, Soyoon;Joo, Myungsoo;Ha, Ki-Tae;Baek, Nam-In;Park, Cheon-Seok;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.1916-1924
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    • 2017
  • In this study, we synthesized a glycosylated derivative of caffeic acid phenethyl ester (CAPE) using the amylosucrase from Deinococcus geothermalis with sucrose as a substrate and examined its solubility, chemical stability, and anti-inflammatory activity. Nuclear magnetic resonance spectroscopy showed that the resulting glycosylated CAPE (G-CAPE) was the new compound caffeic acid phenethyl ester-4-O-${\alpha}-{\small{D}}$-glucopyranoside. G-CAPE was 770 times more soluble than CAPE and highly stable in Dulbecco's modified Eagle's medium and buffered solutions, as estimated by its half-life. The glycosylation of CAPE did not significantly affect its anti-inflammatory activity, which was assessed by examining lipopolysaccharide-induced nitric oxide production and using a nuclear factor erythroid 2-related factor 2 reporter assay. Furthermore, a cellular uptake experiment using high-performance liquid chromatography analysis of the cell-free extracts of RAW 264.7 cells demonstrated that G-CAPE was gradually converted to CAPE within the cells. These results demonstrate that the glycosylation of CAPE increases its bioavailability by helping to protect this vital molecule from chemical or enzymatic oxidation, indicating that G-CAPE is a promising candidate for prodrug therapy.

(1) Studies on the Determination of the Rate Control Steps in the Various Metabolic Cycles (I) - The Affect of the Light to the Enzyme Activities in the Cotyledons of the Germinating Seeds - (대사과정(代謝過程)에 있어서 율속단계(律速段階) 결정(決定)에 관(關)한 연구(硏究) (I) - 발아종자자엽(發芽種子子葉)에 있어서 산소활성(酸素活性)에 미치는 광선(光線)의 영향(影響) -)

  • Sin, Gwi-Nam
    • Applied Biological Chemistry
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    • v.3
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    • pp.1-7
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    • 1962
  • In order to study the rate control step in the protein metabolic course of the chlorophyll formation, the transaminase activities which are obtained freely in the extracts of cotyledons of germinating peas at the light and the dark places, are measured in Beckman mopel D.U, spectrophoto meter at 490 mu. In this case, of two enzymatic reaction products; oxalacetic acid and pyruvic acid, the former is converted to pyruvic acid by aniline citrate and after each pyruvate phenyl hydrazones are extracted by toluenes: when this is treated with strong alcoholic alkali, a colored hydrazone is formed and it is measured by above apparatus. The estimated G.O.T. and G.P.T. in the germinated cotyledons at dark and light places considerably differ in their activities; G.O.T. and G.P.T. activities which are formed at the light are more increased than at the dark and also they differ in their rates through germination, though G.O.T. activity increment is smoothly but that of G.P.T. is more sharply, and they are considered to be directly affected to the chlorophyll formation and indirectly to the growth. G.O.T. and G.P.T. in each fractions of cell in the cotyledons should be formed by dissociation of zymogens in the microsomal fractions and it seems to promoted by light. In the formation of the chlorophyll, the protein metabolism occurred mainly in the microsomal fractions and the rate determining step is found at the point where the zymogene that is able to produce G.P.T. is activated, and this activation is promoted by light as noted above.

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Evaluation of Effective MMP Inhibitors from Eight Different Brown Algae in Human Fibrosarcoma HT1080 Cells

  • Bae, Min Joo;Karadeniz, Fatih;Ahn, Byul-Nim;Kong, Chang-Suk
    • Preventive Nutrition and Food Science
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    • v.20 no.3
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    • pp.153-161
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    • 2015
  • Matrix metalloproteinases (MMPs) are crucial extracellular matrices degrading enzymes that have important roles in metastasis of cancer progression as well as other significant conditions such as oxidative stress and hepatic fibrosis. Marine plants are on the rise for their potential to provide natural products that exhibit remarkable health benefits. In this context, brown algae species have been of much interest in the pharmaceutical field with reported instances of isolation of bioactive compounds against tumor growth and MMP activity. In this study, eight different brown algae species were harvested, and their extracts were compared in regard to their anti-MMP effects. According to gelatin zymography results, Ecklonia cava, Ecklonia bicyclis, and Ishige okamurae showed higher inhibitory effects than the other samples on MMP-2 and -9 activity at the concentrations of 10, 50, and $100{\mu}g/mL$. However, only I. okamurae was able to regulate the MMP activity through the expression of MMP and tissue inhibitor of MMP observed by mRNA levels. Overall, brown algae species showed to be good sources for anti-MMP agents, while I. okamurae needs to be further studied for its potential to yield pharmaceutical molecules that can regulate MMP-activity through cellular pathways as well as enzymatic inhibition.