• Title/Summary/Keyword: 3-10 kDa fraction

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Characteristics of Angiotensin Converting Enzyme Inhibitory Peptides from Thermolysin Hydrolysate of Manila clam, Ruditapes philippinarum Proteins (바지락 단백질 Thermolysin 가수분해물의 Angiotensin Converting Enzyme 저해 Peptide의 특성)

  • Lee Tae Gee;Yeum Dong Min;Kim Seon Bong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.5
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    • pp.529-533
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    • 2002
  • The peptides inhibiting angiotensin converting enzyme (ACE) were isolated from the hydrolysate of manila clam (Ruditapes philippinamm) proteins prepared with thermolysin. The thermolysin hydrolysate was pretreated with membrane filter (MW cut-off 10,000) to obtain the peptide fraction with ACE inhibition. The crude peptides were applied to a Sephadex LH-20 column and eluted with $30\%$ methanol. The three active fractions (A, B and C) were collected and concentrated, and then applied to a SP-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The four active fractions (A-1, A-2, B-1 and C-1) were collected and concentrated, and then applied to a SuperQ-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The maximum inhibitory activity was observed in the fraction B-1Q showed the IC_{50} values of 0.748 $\mu$g. The abundant amino acids obtained from active fraction B-1Q were leucine, isoleucine, alanine and threonine.

Isolation and Characterization of Immunomodulatory Glycoprotein from the Root of Panax ginseng

  • Shin, Han-Jae;Park, Kyeong-Mee;Kim, Young-Sook;Nam, Ki-Yeul;Lee, You-Hui;Park, Jong-Dae
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.128-133
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    • 2000
  • A high molecular (more than 10 kDa) fraction, showing mitogenic and comitogenic activities in spleen cells of mouse, was isolated from water extract of ginseng. The crude protein substance prepared by 80% (NH$_4$)$_2$SO$_4$ precipitation from this fraction was purified and isolated by DEAE Sepharose column chromatography. Among the fractions eluted, it was found that four kinds of fractions eluted with 0 to 1 M NaCl gradient were glycoproteins, which induced proliferation of spleen cells and increased NO production in macrophages. Among them, F-2 fraction, which contained 35.9% protein,49.4% neutral sugar and 12.5% uronic acid, was found to show mitogenic activity as strong as that of LPS (lipopolysaccharide) at a concentration of 100 $\mu\textrm{g}$/ml and to remarkably stimulate NO production by murine macrophages at a concentration of 500 $\mu\textrm{g}$/ml. When F-2 is deproteinized, the mitogenic activity of F-2 was decreased significantly to 70.9% as compared with that of F-2. This results suggests that the protein moiety of F-2 may play an important role in immunomodulating activity of glycoprotein from the root of Panax ginseng.

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Metabolomic Analysis of Ethyl Acetate and Methanol Extracts of Blueberry (Ethyl Acetate와 Methanol을 이용한 블루베리 추출물 대사체 분석)

  • Jo, Young-Hee;Kim, Sugyeong;Kwon, Da-Ae;Lee, Hong Jin;Choi, Hyung-Kyoon;Auh, Joong-Hyuck
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.419-424
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    • 2014
  • Metabolite profiling of blueberry (cultivar "Spartan") was performed by extraction using different solvents, methanol and ethyl acetate, through metabolomic analysis using LC-MS/MS. Unsupervised classification method (PCA) and supervised prediction model (OPLS-DA) provided good categorization of metabolites according to the extraction solvents. Metabolites of the anthocyanin family, including delphinidin hexoside, delphinidin, 5-O-feruloylquinic acid, malvidin hexoside, malvidin-3-arabinoside, petunidin-3-arabinoside, and petunidin hexoside, were mainly detected in methanol fractions, whereas those of the flavonoid family, including chlorogenic acid, chlorogenic acid dimer, 6,8-di-C-arabinopyranosyl-luteolin, and luteolin were successfully prepared in the ethyl acetate fraction. Thus, metabolomic analysis of blueberry extracts allows for the simple profiling of whole and distinctive metabolites for future applications.

Optimization of Whey-Based Medium for Growth and ACE-Inhibitory Activity of Lactobacillus brevis

  • Ahn, Jae-Eun;Park, Seung-Yong;Lee, Byong-H.
    • Journal of Dairy Science and Biotechnology
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    • v.25 no.1
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    • pp.1-7
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    • 2007
  • A Whey-based medium was formulated with Lactobacillus brevis to investigate whether any functional peptides could derive from whey protein. The optimal concentrations of the ingredients of the medium for the growth of Lactobacillus were determined as 2% whey protein concentrate and 1% glucose and 0.5% yeast extracts. The growth of Lb. brevis was improved with the supplementation of yeast extracts than glucose. The viable cells counts of Lb. brevis reached to 2.0 × 10$^8$CFU/mL in the whey-based medium. The whey protein hydrolysates recovered from the supernatant after centrifugation at 10,000 x g for 10min induced strong inhibitory activity against ACE. When the whey protein hydrolysate were partially purified by a membrane tubing below 8,000Da, the partially purified fraction remained 64.7 ${\pm}$ 3.6% of the ACE inhibition activity of the whey protein hydrolysates and IC$_{50}$ was 38.8 ${\pm}$ 2.2mg/mL. The whey-based medium was proved to be effective in producing ACE inhibitory peptides by lactic bacteria fermented whey protein.

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Probiotics with Antimicrobial Activity against Multidrug Resistant Pseudomonas aeruginosa and Acinetobacter baumannii (다제내성 녹농균과 아시네토박터 바우마니에 항균활성을 가지는 프로바이오틱스)

  • Lee, Do Kyung;Kim, Min Ji;Kang, Joo Yeon;Park, Jae Eun;Shin, Hea Soon;Ha, Nam Joo
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.245-252
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    • 2013
  • Pseudomonas aeruginosa and Acinetobacter baumannii are significant opportunistic pathogens in hospitals and are resistant to most antibiotics. Multidrug-resistant P. aeruginosa (MDRPA) and A. baumannii (MDRAB) cause severe human nosocomial infections and are more difficult to treat than methicillin-resistant Staphylococcus aureus (MRSA). Bifidobacteria are among of the most beneficial probiotics and have been widely studied for their antimicrobial activities. The present study explored the antimicrobial activity of Bifidobacterium sp. isolated from healthy Koreans against MDRPA and MDRAB. The antimicrobial activity of the isolates against MDRPA and MDRAB, which are resistant to ciprofloxacin, tobramycin, gentamicin, meropenem, and ceftazidime, was determined by modified broth microdilution methods using absorbance. Among all tested bifidobacteria isolates (nine B. adolescentis, three B. longum, and two B. pseudocatenulatum), the culture supernatant of B. pseudocatenulatum SPM1309 showed a strong growth inhibitory effect against MDRPA and MDRAB. No change in the turbidity of the mixture was observed during incubation, and its inhibitory effect occurred through bacteriostastic action. Moreover, the antibacterial activity was observed in the fraction with molecular weights <10 kDa of bifidobacteria culture supernatant, and the active fraction was heat-stable because it maintained its activity when heated at $70^{\circ}C$ for 10 min. The results suggest that this Bifidobacterium strain could have potential applications for alternative therapy in MDRPA and MDRAB infections.

Isolation of Polysaccharides Modulating Mouse’s Intestinal Immune System from Peels of Citrus unshiu (귤피로부터 분리한 마우스의 장관면역 활성 다당류의 검색)

  • Yang, Hyun-Seuk;Yu, Kwang-Won;Choi, Yang-Mun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1476-1485
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    • 2004
  • Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

Antioxidant and Cytoprotective Effects of Lotus (Nelumbo nucifera) Leaves Phenolic Fraction

  • Lee, Da-Bin;Kim, Do-Hyung;Je, Jae-Young
    • Preventive Nutrition and Food Science
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    • v.20 no.1
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    • pp.22-28
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    • 2015
  • Phenolic rich ethyl acetate fraction (EAF) from lotus leaves was prepared and its bioactive components, antioxidant and cytoprotective effects were investigated. EAF showed high total phenolic content and flavonoid content and contained rutin ($11,331.3{\pm}4.5mg/100g\;EAF$), catechin ($10,853.8{\pm}5.8mg/100g\;EAF$), sinapic acid ($1,961.3{\pm}5.6mg/100g\;EAF$), chlorogenic acid ($631.9{\pm}2.3mg/100g\;EAF$), syringic acid ($512.3{\pm}2.5mg/100g\;EAF$), and quercetin ($415.0{\pm}2.1mg/100g\;EAF$). EAF exerted the $IC_{50}$ of $4.46{\mu}g/mL$ and $5.35{\mu}g/mL$ toward DPPH and ABTS cation radicals, respectively, and showed strong reducing power, which was better than that of ascorbic acid, a positive control. Additionally, EAF protected hydroxyl radical-induced DNA damage indicated by the conversion of supercoiled pBR322 plasmid DNA to the open circular form and inhibited lipid peroxidation of polyunsaturated fatty acid in a linoleic acid emulsion. In cultured hepatocytes, EAF exerted a cytoprotective effect against oxidative stress by inhibiting intracellular reactive oxygen species formation and membrane lipid peroxidation. In addition, depletion of glutathione under oxidative stress was remarkably restored by treatment with EAF. The results suggest that EAF have great potential to be used against oxidative stress-induced health conditions.

Antioxidant and Cellular Protective Activities of Ecklonia cava Extracts against Reactive Oxyegen Species (감태(Ecklonia cava) 추출물의 항산화 및 세포보호 활성)

  • Yoo, Cha Young;Kim, Si Yun;Park, Jung Won;Sung, Soo An;Kim, Da Ae;Park, Jee Hyun;Xuan, Song Hua;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.3
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    • pp.287-294
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    • 2015
  • In this study, we investigated the antioxidative effects of brown seaweed Ecklonia cava extract and its subfractions. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of E. cava. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of ethyl acetate fraction ($FSC_{50}=6.98{\mu}g/mL$) and aglycone fraction ($7.03{\mu}g/mL$) are similar to that of (+)-${\alpha}$-tocopherol ($8.98{\mu}g/mL$) which is a reference control. Reactive oxygen species (ROS) scavenging activity (total antioxidant capacity, $OSC_{50}$) of the aglycone fraction ($OSC_{50}=14.48{\mu}g/mL$) on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay was the strongest among all extract and fractions. However, all samples showed lower antioxidant activities than that of L-ascorbic acid ($6.88{\mu}g/mL$) known as a powerful antioxidant. The protective effect of 50% ethanol extract on the $^1O_2$-induced cellular damage of human erythrocytes was dependent on the concentration from 5 to $50{\mu}g/mL$. Both ethyl acetate fraction and aglycone fraction showed strong cellular protective activities at $10{\mu}g/mL$, where the cellular protective effects (${\tau}_{50}$) of each fraction were recorded 442.0 min and 539.9 min, respectively. Three kinds of extract/fractions of E. cava showed much greater cellular protective activities at $10{\mu}g/mL$ than that of liposoluble antioxidant (+)-${\alpha}$-tocopherol (40.6 min) which is a reference control. These results suggest E. cava extracts and its fractions can be applied as an antioxidant ingredient in a field of cosmetics.

Deastringent Peel Extracts of Persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) Protect Neuronal PC-12 and SH-SY5Y Cells against Oxidative Stress

  • Jeong, Da-Wool;Cho, Chi Heung;Lee, Jong Suk;Lee, Seung Hwan;Kim, Taewan;Kim, Dae-Ok
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1094-1104
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    • 2018
  • The peel of astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) is a by-product of dried persimmon (gotgam). We investigated if deastringent peel extracts of persimmon cv. Cheongdo-Bansi had antioxidative and neuroprotective properties. Two different extracts were prepared: thermally and nonthermally treated persimmon peel extracts (TPE and NTPE, respectively). Both TPE and NTPE were fractionated sequentially in n-hexane, chloroform, ethyl acetate, n-butanol, and water. The TPE and NTPE ethyl acetate fractions had the highest total phenolic and flavonoid contents as well as antioxidant capacities among all the fractions. Pretreatment of neuronal PC-12 and SH-SY5Y cells with the TPE and NTPE ethyl acetate fractions increased cell viability after exposure to oxidative stress. The ethyl acetate fraction of TPE attenuated oxidative stress inside both PC-12 and SH-SY5Y cells more effectively than that of NTPE. Furthermore, the TPE and NTPE ethyl acetate fractions inhibited acetylcholinesterase and butyrylcholinesterase. Analysis of ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry results revealed gallic acid, kaempferol, kaempferol-3-O-galactoside, kaempferol-3-O-glucoside, quercetin, quercetin3-O-galactoside, quercetin-3-O-galactoside-2'-O-gallate, and quercetin-3-O-glucoside as the major phenolics of the TPE and NTPE ethyl acetate fractions. Taken together, these results suggest that the ethyl acetate fraction of deastringent persimmon peel is rich in antioxidants and has potential as a functional food to reduce oxidative stress.

Codon usage analysis of rice glutelin genes (쌀 저장 단백질 글루텔린 유전자 암호 분석)

  • Shin, Yun-Cheol;Kim, Ju-Kon;Nahm, Baek-Hie
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.517-524
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    • 1993
  • To characterize glutelins, the most abundant storage protein in rice, 13 complete coding sequences of glutelin genes from the database were analyzed. According to the phylogenic analysis, these genes could be classified into 5 groups, Group I to V. The degrees of homology were calculated to be in the range of 90 to 60%, but the patterns of hydrophobicity were similar in all the groups. Also, each group was found to have similar amino acid composition with variations in lysine content from 2.5 to 3.6% due to the point mutation of arginine to lysine. The isoelectric points of mature proteins and their basic chains of all the groups showed the value of about 9.0 and 10.0, respectively, while the isoelectric points of acidic chains in these groups showed the distinct value of 6.6, 6.7, 7.2, 8.4 and 7.9. The plot of the fraction of G+C at synonimous site in codons (GC3s) against effective codon numbers suggest no major difference in translational efficiency in the expression of glutelin multigenes.

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