• Asian-Australasian Journal of Animal Sciences
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• v.22 no.11
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• pp.1587-1593
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• 2009

Duck egg white (DEW) hydrolysates were prepared by five enzymes (papain, trypsin, chymotrypsin, alcalase, and flavourzyme) and their antioxidant activities investigated in this study. DEW hydrolyzed with papain (DEWHP) had the highest peptide content among the five enzymatic treatments. Besides, the peptide content of DEWHP increased when the enzyme to substrate ratio (E/S ratio) increased. It was suggested that higher E/S ratio contributed to elevate the degree of hydrolysis of DEW effectively. Similar results were also obtained by Tricine-SDS-PAGE. In addition, SDS-PAGE patterns indicated papain was the only one amongst all enzymes with the ability to hydrolyze DEW. In antioxidant properties, DEWHP showed more than 70% of inhibitory activity on linoleic acid peroxidation and superoxide anion scavenging. Moreover, the $Fe^{2+}$ chelating effect of DEWHP was greater than 90%, while no significant difference was observed in DPPH radical scavenging and reducing ability. The results of peptide contents, antioxidant activities and electrophoresis suggested that the higher the peptide content, the stronger the antioxidant activities in DEWHP.

• Preventive Nutrition and Food Science
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• v.20 no.1
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• pp.67-72
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• 2015

Squid is one of the most important commercial fishes in the world and is mainly utilized or consumed as sliced raw fish or as processed products. The biofunctional activities of enzymatic squid meat hydrolysate were determined to develop value-added products. Enzymatic squid hydrolysate manufactured by Alcalase effectively quenched 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, and hydrogen peroxide radical with $IC_{50}$ values of 311, 3,410, and $111.5{\mu}g/mL$, respectively. Angiotensin I-converting enzyme inhibitory activity of squid hydrolysate was strong with an $IC_{50}$ value of $145.1{\mu}g/mL$, while tyrosinase inhibitory activity with an $IC_{50}$ value of 4.72 mg/mL was moderately low. Overall, squid meat hydrolysate can be used in food or cosmetic industries as a bioactive ingredient and possibly be used in the manufacture of seasoning, bread, noodle, or cosmetics.

• Journal of Marine Bioscience and Biotechnology
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• v.10 no.1
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• pp.1-8
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• 2018

The objective of this study was to purify and characterize the ${\beta}-secretase$ inhibitor from enzymatic hydrolysates of blackfin flounder muscle, for development of a novel anti-dementia agent that may be used in the drug or functional food industries. ${\beta}-secretase$ inhibitory peptide was purified from various enzymatic hydrolysates of blackfin flounder muscle. Among six enzymatic hydrolysates, the Alcalase hydrolysate revealed highest ${\beta}-secretase$ inhibitory activity. Consecutive purification of the blackfin flounder muscle hydrolysate using Sephadex G-25 column chromatography and octadecylsilane C18 reversed phase HPLC techniques were used to isolate a potent ${\beta}-secretase$ inhibitory peptide composed of 5 amino acids, Leu-Thr-Gln-Asp-Trp (MW: 526.7 Da). The $IC_{50}$ value of purified ${\beta}-secretase$ inhibitory peptide was $126.93{\mu}M$. Results of this study suggest that peptides derived from blackfin flounder muscle may be beneficial as anti-dementia compounds in functional foods or as pharmaceuticals.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.5
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• pp.456-463
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• 2011

We attempted to isolate a collagenase-1 inhibitory peptide from skate Dipturus chilensis skin protein. The protein from skate skin was digested by various enzymes (alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin) to produce a collagenase-1 inhibitory peptide. The collagenase-1 inhibitory activity of the peptides obtained was measured by gelatin digestion assay. Among the six hydrolysates, pepsin hydrolysate exhibited the highest collagenase-1 inhibitory activity. The peptide showing strong collagenase-1 inhibitory activity was purified by Sephadex G-25 gel chromatography and HPLC using an octadecylsilyls (ODS) column. The amino acid sequence of purified collagenase-1 inhibitory peptide was identified to be Asn-Leu-Asp-Val -Leu-Glu-Val-Phe (961 Da) by quadrupole time of flight (Q-TOF) and electrospray ionization mass spectrometry (ESI-MS) mass spectroscopy. The $IC_{50}$ value of purified peptide was 87.0 ${\mu}M$. Moreover, the peptide did not exhibit cytotoxic effects on human dermal fibroblast cell lines.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.594-599
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• 2016

Infection with HIV (Human immunodeficiency virus), over time, develops into acquired immunodeficiency syndrome (AIDS). The development of non-toxic and effective anti-HIV drugs is one of the most promising strategies for the treatment of AIDS. In this study, we investigated the anti-HIV-1 activity of gelatin hydrolysates from Alaska pollack skin. Gelatin hydrolysates were prepared using four enzymes (alcalase, flavourzyme, neutrase, and pronase E). Among these, the pronase E gelatin hydrolysate was found to inhibit HIV-1 infection in the human T cell-line MT4. It exhibited inhibitory activity on HIV-1_IIIB-induced cell lysis, reverse transcriptase activity, and viral p24 production at noncytotoxic concentrations. Moreover, it decreased the activation of matrix metalloproteinase-2 (MMP-2) in vitro. Because HIV infection-induced activation of MMP-2 can accelerate collagen resolution and collapse of the immune system, pronase E gelatin hydrolysate might prevent the activation of MMP-2 in cells, resulting in collagen stabilization and immune cell homeostasis consistent with anti-HIV activation. These results suggest that pronase E gelatin hydrolysate could potentially be incorporated into a novel therapeutic agent for HIV/AIDS patients.

• ALGAE
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• v.25 no.1
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• pp.45-56
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• 2010

In this study, we assessed the antioxidant properties of tidal pool microalgae, Halochlorococcum porphyrae and Oltamannsiellopsis unicellularis, from Jeju Island, Korea. Specifically, the antioxidant activity of fractions isolated from 80% methanol extract, and digests produced from five proteases and carbohydrases, were investigated. Almost all the fractions and the 80% methanol extract exhibited higher effects on 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging. The ethyl acetate fraction showed the highest superoxide anion scavenging activity, while both n-hexane and chloroform fractions exhibited higher $H_2O_2$ scavenging activity. Among the enzymatic digests from H. porphyrae and O. unicellularis, all the digests exhibited remarkable DPPH scavenging activities. In nitric oxide inhibition, all the digests recorded significantly higher effects than those of the commercial antioxidants (p < 0.05). Flavozyme and Neutrase digests from H. porphyrae, and Termamyl and Alcalase digests from O. unicellularis, showed significant effects in metal chelating. Lipid peroxidation was significantly inhibited in the ethyl acetate fraction, in the Celluclast and Protamex digests from H. porphyrae, and in the chloroform fraction from O. unicellularis. These findings suggest that the two tidal pool microalgae tested in this study are rich in potential antioxidative compounds, the specific properties of which can be considered for use in the food and pharmaceutical industries.

• Food Science and Biotechnology
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• v.14 no.5
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• pp.614-620
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• 2005

The antioxidative effect of Ecklonia cava, a brown marine alga, was investigated on radical scavenging, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), and hydroxyl and alkyl radicals, using an electron spin resonance (ESR) technique, and on the inhibition of $H_2O_2$-induced DNA damage using comet assay. E. cava was enzymatically hydrolyzed with five food industrial proteases (Alcalase, Flavourzyme, Kojizyme, Neutrase and Protamex) to prepare water-soluble extracts. All the proteolytic hydrolysates exhibited strong dose-dependent radical scavenging activities (above 80%) at a concentration of $2.5\;{\mu}g/mL$. Kojizyme extract (obtained by proteolytic hydrolysation of E. cava with Kojizyme) showed the highest hydroxyl radical scavenging activity of around 98%. In addition, the $H_2O_2$-induced DNA damage was determined using a comet assay, which was quantified by measuring the tail length. Reduction of DNA damage increased with increasing concentrations of Kojizyme extract from E. cava. These results indicated that E. cava has a potential as a valuable natural antioxidative source.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.2
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• pp.118-125
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• 2011

In this study, an angiotensin I-converting enzyme (ACE) inhibitor from squid skin was purified and characterized. Squid (Todarodes pacificus) skin protein isolates were hydrolyzed using six commercial proteases: alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin. The peptic hydrolysate had the highest ACE inhibitory activity. The ACE inhibitory peptide was purified using Sephadex G-25 column chromatography and reverse phase high-performance liquid chromatography (HPLC) with a $C_{18}$ column. The purified ACE inhibitory peptide was identified and sequenced, and found to consist of seven amino acid residues: Ser-Ala-Gly-Ser-Leu-Val-Pro (657Da). The $IC_{50}$ value of the purified ACE inhibitory peptide was 766.2 ${\mu}M$, and Lineweaver-Burk plots suggested that the purified peptide acts as a noncompetitive ACE inhibitor. These results suggest that the ACE inhibitory peptide purified from the peptic hydrolysate of squid skin may be of benefit in developing antihypertensive drugs and functional foods.

• Preventive Nutrition and Food Science
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• v.13 no.3
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• pp.166-175
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• 2008

In this study, we focused on natural water-soluble antioxidants from the Jeju benthic diatoms, Achnanthes longipes, Navicula sp. and Amphora coffeaeformis. They were prepared by enzymatic digestion using five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase) and their potential antioxidant activity was assessed. Among the enzymatic digests, Neutrase digest from A. coffeaeformis exhibited the highest effect in DPPH radical scavenging. Flavourzyme (48.7%), Viscozyme (47.4%) and Celluclast (45.7%) digests from Navicula sp. exhibited higher $O^{{\cdot}-}_2$ radical scavenging activity. Viscozyme digest from A. coffeaeformis (45.9%) possessed the highest effects in hydroxyl radical scavenging. Termamyl (89.3%) and Protamex (88.8%) digests from A. coffeaeformis had strong metal chelating activity. Lipid peroxidation was significantly inhibited in Termamyl and Kojizyme digests from A. longipes, AMG and Termamyl digests from Navicula sp. and Kojizyme digest from A. coffeaeformisi. These data suggest that enzymatic digests of the Jeju benthic diatoms might be valuable sources of antioxidant which can be applied in food and pharmaceutical industry.

• KSBB Journal
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• v.15 no.6
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• pp.635-643
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• 2000

To compare the antioxidative activities of fish skin and bovine skin gelatin hydrolysate, gelatin hydrolysates from Alaska pollack and bovine skin were prepared by various enzymatic hydrolysis methods (1st step, Alcalase; 2nd step, pronase E; 3rd step, collagenase) using a continuous three-step membrane reactor. The molecular weight distributions of the 1st, 2nd and 3rd step hydrolysates were 7∼10 kDa, 2∼5 kDa and 0.7∼0.9 kDa, respectively. The antioxidative activity of fish skin gelatin hydrolysate was stronger than that of bovine skin gelatin hydrolysate, and in particular, both of 2nd step hydrolysates showed more antioxidative activity than hydrolysates of any other step. The optimum antioxidative activity concentration of the 2nd step hydeolysates of fish and boving skin were 1% (w/w) in a linoleic acid water-alcohol emulsion. In cultured cells exposed to t-butyl hydroperoxide (t-BHP), the 2nd step hydrolysate of fish skin gelatin delayed cell death most. These results suggest that the antioxidative activity of fish skin gelatin hydrolysate is higher than that of bovine skin gelatin hydrolysate because of their different amino acid contents.