• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.466-472
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• 1997

Peptides separated from fish paste washing liquid of an Alaska pollak (Theragria chalcogramma) were purified and characterized. The fish paste washing liquid (supernatant) was separated by centrifugation of fish paste homogenate. The fish paste washing liquid of $0.5\%$ concentration was hydrolyzed for 24 hour at $50^{\circ}C$ by immobilized protease in bioreactor and decomposing liquid of protein having $50\%$ decomposing rate (OPA method) was obtained. The crude peptide fractions were obtained from this liquid by Dowex 50w $(H^+)$ column chromatograpy. Purified peptides (SP-fraction peptides) were fractionated by using SP-Sepadex C-25 $(H^+)$ column chromatography. Molecular weights and amino acid compositions of these peptides were estimated by Sephadex G-50 column chromatography and HPLC, respectively. when the washed peptides was eluated with $0.6\~0.9\%\;and\;1.2\~2.0\%$ of NaCl, peptides composed of weakly basic amino acids and strongly basic amino acid were respectively eluted. Molecular weights of each peptide fractions showed the broad distribution from 1,000 Da to 3,000 Da in the order of SP-4>SP-3>SP-2>SP-1. Peptides contained a large quantity of glycine, arginine, glutamic acid, and alanine in the washed peptide and its SP-tractions, respectively.

• KSBB Journal
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• v.29 no.5
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• pp.380-384
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• 2014

Inhibition effects of peptide hydrolysates from Astragalus membranaceus Bunge. on the expression of the matrix metalloproteinases (MMPs) in human dermal fibroblasts were evaluated in vitro. Crude peptides were obtained by the hydrolysis of proteins extracted from A. membranaceus. Peptides were purified partially by the basis on the molecular weight using 40% polyacrylamide gel electrophoresis before treatment with human dermal fibroblasts. Basis on the doseeffect experiments, expressions of MMPs including MMP-1, MMP-3, MMP-8, MMP-13 in human dermal fibroblasts were evaluated. Expressions of MMP-1, MMP-3, MMP-8 and MMP-13 were reduced in 43%, 5%, 22% and 57% respectively. The mass spectrometric analysis of partially purified peptides from A. membranaceus, which strongly inhibit expressions of MMPs, indicated that the peptides were composed of molecules below 1500 Da.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.12
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• pp.5139-5145
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• 2016

There has been an increment in the number of studies focused on marine bioactive materials. Many peptides and other biomaterials with anticancer potential have been extracted from various marine animals. Artemia extracts have found uses in sun-light protection cosmetics and anti-aging products. However, contents of biochemical compounds in Artemia spp. and molecular mechanisms of have not been clearly studied in leukemic cells in vitro. In this work, we isolated and purified proteins of Artemia Urmiana. Six clear fractions (A-F) observed on DEAE-cellulose chromatography were assayed for effects on cell growth, differentiation and apoptosis using the human leukemic HL-60 cell line. Cell proliferation analysis by MTT and BrdU assays indicated that did not affect cells, growth. Cells treated with crude extract and fractions A, B and C, but not E and F (up to $100{\mu}g/mL$), exhibited increase of cell growth in a dose dependent manner. Stimulatory effects of fraction D were observed at concentrations of $10{\mu}g/mL$ and above. In nitro blue tetrazolium (NBT) reduction assays, treatment with $100{\mu}g/mL$ of fraction E or F for 96 hr increased the fraction of differentiated cells up to $14.8{\pm}3.56%$ and $16.5{\pm}2.08%$ respectively. Combination of those fractions with retinoic acid had significant synergistic effects on the differentiation of cells ($56.8{\pm}3.7%$ and $67.4{\pm}4.2%$, $p{\leq}0.01$). Annexin-V FITC staining for apoptosis and flow cytometric assays indicated induction of apoptosis by fractions E and F up to 23.8 and 31.8% of cells.

• Journal of the Korean Chemical Society
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• v.42 no.2
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• pp.214-219
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• 1998

Dynorphin B analogues, $[Arg^{11}, D-Ala^{12}]$dynorphin B, $[D-Ala^2, Ala^6, Arg^{11}, D-Ala^{12}]$dynorphin B, and dynorphin B (1-11) were synthesized by solid-phase method. A chloromethylated polystyrene resin cross-linked with 2% divinylbenzene was substituted with Thr in ethanol to contain 1.20 mmol Thr/g of resin. All amino groups of amino acids were protected with t-Boc group and 2,6-dichlorobenzyl and nitro groups were used to protect the side chains of Tyr and Arg, respectively. Stepwise synthetic method was applied for synthesis of the products. Dicyclohexylcarbodiimide (DCC) and 1-hydroxybenzotriazole (HOBT) were used as the coupling reagents. The crude peptides were purified by gel filteration on Sephadex LH-20 column $(2 \times 50 cm)$ using MeOH/MeCN (3/1) and then characterized with HPLC, amino acid analyzer.

• Korean Journal of Food Science and Technology
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• v.37 no.5
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• pp.827-832
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• 2005

Bacillus subtilis JM3 protease from naturally fermented anchovy sauce was partially purified in 40-60% ammonium sulfate fraction. To accelerate fermentation of anchovy sauce, 2 and 4% crude B. subtilis JM3 proteases were added to 6 month-ripened anchovy sauce, and their hydrolysis degrees and amino-nitrogen contents were investigated at different storage times. Low molecular weight (LMW) peptide was purified by ultrafiltration ana gel permeation chromatography from anchovy sauce manufactured with B, subtilis JM3 protease. Anchovy sauces with 2 and 4% proteases increased hydrolysis rate by 27 and 32%, respectively. Amino-nitrogen contents of anchovy sauces fermented with 2 and 4% proteases were twofold higher than that of control. Control showed five peptide peaks on Bio-Rad P2 gel permeation chromatography spectrum, whereas anchovy sauces with 2 and 4% B. subtilis JM3 proteases showed six and seven peaks, respectively. ACE inhibitory activity was highest in peak 6 (43.75%) of anchovy sauce with 2% protease, followed by peak 5 (34.82%) of control. DPPH radical-scavenging effect was higher than 50% in all samples. Cytotoxicity was highest in peak 3 (44.12%) of control, fellowed by peak 5 (42.04%) of anchovy sauce with 4% protease.