The Antioxidant and Anti-inflammatory Effects of Abalone Intestine Digest, Haliotis discus hannai in RAW 264.7 Macrophages

천충길,Sun-Ae Kim,이준식,김학주,최일환,정원교 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.3

Abalone is a marine gastropod and an important fishery and food industrial resource that is massively maricultured in Asia, Africa, Australia, and America. However,the health beneficial effects of abalone have rarely been reported. In the present study, we examine the antioxidant and anti-inflammatory effects of abalone Haliotis discus hannai in macrophage cells. The results showed that abalone intestine digest (AID) has antioxidant activities against lipid peroxidation, ROS stress and DNA damage in H2O2-treated RAW264.7 macrophages. In the lipopolysaccharide (LPS)-induced macrophages, AID suppresses LPS-induced production of nitric oxide (NO) via inducible nitric oxide synthase (iNOS) expression in a dose-dependent manner. It also significantly reduced the generation of proinflammatory cytokines, such as interleukin (IL-1β),tumor necrosis factor (TNF)-α, and IL-6. Furthermore, AID significantly suppresses the phosphorylation of mitogen-activated protein kinases (MAPKs) such as ERK,JNK, and p38. These results indicated that AID inhibits oxidative damage by ROS and LPS-induced inflammatory response via blocking the MAPK signaling pathway in murine macrophages. The potent antioxidant and antiinflammatory effects of abalone intestine as byproducts from fishery manufacturing may suggest the possibility of high valuable utilization and application as a nutraceutical and therapeutic substance.

Isolation and Antioxidant Activity Evaluation of Two New Phthalate Derivatives from Seahorse, Hippocampus Kuda Bleeler

천충길,강경화,김세권 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.5

Seahorse (Hippocampus Kuda Bleeler) has been used as traditional medicine for thousands of years, in Eastern Asia. In this study of the methanol extract of fresh Hippocampus Kuda, the new compounds 2-ethyldecyl 2-ethylundecyl phthalate (1), 2, 12-diethyl-11-methylhexadecyl 2-ethyl-11-methylhexadecylphthalate (2), along with a known Bis(2-ethylheptyl) phthalate (3) were isolated. They were tested for their antioxidant activities, including lipid peroxidation inhibitory activity, DPPH radical scavenging,hydroxyl radical scavenging, superoxide anion radical scavenging, alkyl radical scavenging, and cellular radicals;these can be detected using a fluorescence probe, 2’,7’-dichlorofluorescin diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF)with the presence of intracellular ROS on mouse macrophages,RAW264.7 cell. Compound (2) exhibited the highest antioxidant activity and inhibitory intracellular ROS than another compounds (1, 3). Furthermore, MTT assay showed no cytotoxicity on mouse macrophages cell (RAW264.7) and human fetal lung fibroblast cell line (MRC-5). This antioxidant property depends on concentration and increasing with increased amount of the compound.

Isolation and Characterization of Collagen from Skin of Bullfrog, Rana catesbeiana Shaw

천충길,정원교,Ngo D. Nghiep,이상훈,김세권 한국수산과학회 2007 Fisheries and Aquatic Sciences Vol.10 No.2

Free Radical and Reactive Oxygen Species Scavenging Activities of the Extracts from Seahorse, Hippocampus kuda Bleeler

천충길,류보미,김문무,김세권 한국생물공학회 2008 Biotechnology and Bioprocess Engineering Vol.13 No.6

Seahorse, Hippocampus kuda (SH) a marine teleost fish, is well known not only for its special medicinal composition and used as one of the most famous and expensive materials of traditional Chinese medicine. It was extracted with water (SHW), methanol (SHM), and ethanol (SHE), respectively and evaluated by various antioxidant assays. The including reducing power, total antioxidant, DPPH radical scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging, alkyl radical scavenging, and protective effect on DNA damage caused by hydroxyl radicals generated. Further, the ROS level was detected using a fluorescence probe, 2',7'-dichlorofluorescin diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS on mouse macrophages, RAW264.7 cell and inhibited myeloperoxidase (MPO) activity in human myeloid, HL60 cells, respectively. Those various antioxidant activities were compared to standard antioxidants such as α-tocopherol. Among SHM exhibited the highest antioxidant activity in linoleic acid system, effective reducing power, DPPH radical scavenging, hydroxyl radical scavenging, superoxide radical scavenging, alkyl radical scavenging, inhibitory intracellular ROS, and inhibited MPO activity. Furthermore, MTT assay showed no cytotoxicity on mouse macrophages cell (RAW264.7) and human cell lines (MRC-5, HL60, U937). This antioxidant property depends on concentration and increasing with increased amount of extracts. The results obtained in the present study indicated that the see horse (Hippocampus kuda Bleeker) is a potential source of natural antioxidant.

Characterization of Growth and Protein Contents from Microalgae Navicula incerta with the Investigation of Antioxidant Activity of Enzymatic Hydrolysates

강경화,천충길,유보미,Se-Kwon Kim 한국식품과학회 2011 Food Science and Biotechnology Vol.20 No.1

Microalgae are major primary producers of organic matters in aquatic environments through their photosynthetic activities. Benthic diatom Navicula incerta is the major component of phytoplankton and also relatively easy to cultivate, used as live food source in aquaculture. The growth characteristics of N. incerta were estimated under combinations of temperature, salinity, and light; and also its composition and antioxidant activities were determined. The maximum cell density of 87×10^5cells/mL, was reached at 20^oC, 250 μmol/m^2·sec, 33‰ salinity,pH 8.3, 12:12 light:dark, and F/2 medium on 2 weeks of the culture period. The antioxidant enzymatic hydrolysates efficiently quenched different free radicals: 1,1-diphenyl-2-pycryl-hydrazyl (DPPH) (pepsin IC_50=196.0 μg/mL), hydroxyl (α-chymotrypsin IC_50=102.0 μg/mL), and superoxide (neutrase IC_50=169.0 μg/mL). These results suggest that the enzymatic hydrolysate from N. incerta acts as a candidate against antioxidant and could be used as a potential functional food ingredient.

Angiotensin I-Converting Enzyme Inhibitory Activity of Sulfated Chitooligosaccharides with Different Molecular Weights

김세권,천충길,엄태길,유보미 한국키틴키토산학회 2010 한국키틴키토산학회지 Vol.15 No.2

The angiotensin I converting enzyme (ACE) inhibitory effects of three kinds of sulfated chitooligosaccharides (SCOS),relatively higher molecular weight SCOS (HMWSCOS, 5-10 kDa), medium molecular weight SCOS (MMWSCOS, 5-3 kDa),and lower molecular weight SCOS (LMWSCOS, 3-1 kDa), respectively. The MMWSCOS exhibited the higher inhibition activity with the IC50 value of 0.25 mg/mL than other sulfated chitooligasaccharides (MMWSCOS: 0.775 mg/mL and LMWSCOS:0.325 mg/ml). Furthermore, all SCOSs showed no cytotoxicity on human embryonic lung fibroblast cell line (MRC-5) and Lineweaver-Burk plots suggest that MMWSCOS and LMWSCOS acts as non-competitive inhibitor to inhibit ACE. Therefore,these results exhibited that substitution of the hydrogen atom at the C-6 position of pyranose residue by the sulfate group promotes ACE inhibitory effects of COS and they would be beneficial ingredients for nutraceuticals and pharmaceuticals against hypertension and related diseases.

Antioxidant Peptide Isolated from Muscle Protein of Bullfrog, Rana catesbeiana Shaw

제재영,천충길,김세권 한국식품영양과학회 2007 Journal of medicinal food Vol.10 No.3

Bullfrog (Rana catesbeianaShaw) muscle protein was enzymatically hydrolyzed for extraction of an antiox-idant peptide. Antioxidant peptide from bullfrog muscle protein hydrolysate (APBMH) was purified using consecutive chro-matographic methods, and the amino acid sequence was identified as being Leu-Glu-Gln-Gln-Val-Asp-Asp-Leu-Glu-Gly-Ser-Leu-Glu-Gln-Glu-Lys-Lys (molecular mass of 1,988 Da) by quantitative time-of-flight electrospray ionization massspectroscopy. To assess antioxidant activities of APBMH, two different in vitrosystems were employed: free radical scav-enging activity by electron spin resonance (ESR) spectroscopy and polyunsaturated fatty acid (PUFA) peroxidation inhibitionassay. ESR revealed that APBMH is an effective free radial scavenger with activity similar to that of vitamin C against 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, and superoxide radicals, and its 50% inhibitory concentration values were 179.4 .M,162.7 .M, and 176.1 .M, respectively. APBMH also significantly retarded PUFA oxidation, and more potently than did .-tocopherol, which was used as a positive control. In addition, the ability of APBMH to inhibit the oxidative damage of DNAwas assessed, in vitro, by measuring the conversion of supercoiled pBR322 plasmid DNA to the open circular form. It wasfound that APBMH significantly protected hydroxyl radical-induced DNA damage dose-dependently.

Free Radical Scavenging Activity of a Novel Antioxidative Peptide Isolated from In Vitro Gastrointestinal Digests of Mytilus coruscus

김세권,정원교,천충길,이상훈,최선영,성낙주,변희국 한국식품영양과학회 2007 Journal of medicinal food Vol.10 No.1

A low-molecular-weight peptide with potent antioxidative activity was obtained from Mytilus coruscusmus-cle protein using an in vitrogastrointestinal digestion system. The potent antioxidant peptide, which was identified as Leu-Val-Gly-Asp-Glu-Gln-Ala-Val-Pro-Ala-Val-Cys-Val-Pro (1.59 kDa), exhibited higher protective activity against polyunsatu-rated fatty acid (PUFA) peroxidation than the native antioxidants, ascorbic acid and .-tocopherol. In a free radical scavengingassay using electron spin resonance spectroscopy, hydroxyl radical formation was quenched by 75.04% in the presence of M.coruscuspeptide (50 .g/mL), which was similar to ascorbic acid. In addition, the purified peptide could also quench super-oxide and carbon-centered radicals, but those activities were weaker than for ascorbic acid. This study showed that the low-molecular-weight peptide released from in vitrogastrointestinal digestion of mussel exhibited potent antioxidant potential byinhibiting the formation of reactive oxygen species formed by the peroxidation of PUFAs.KEY WORDSantioxidative peptidefree radical scavenging activityin vitro gastrointestinal digestionlipid per-oxidation Mytilus coruscus197

Isolation and Biochemical Characterization of Collagens from Seaweed Pipefish, Syngnathus Schlegeli

Sher Bahadar Khan,천충길,유보미,Se-Kwon Kim 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.4

Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the seaweed pipefish (Syngnathus schlegeli) and partially characterized. The amount of collagens isolated in the subsequent treatments was 5.5% of ASC and 33.2% PSC on the basis of lyophilized pipefish body weight, respectively. According to the electrophoretic pattern and CM-cellulose column chromatogram, the collagens might be classified as type I collagens, containing α1 and α2 chain. The imino acid content of collagen from pipefish was lower than those of mammalian collagens as also were the denaturation temperatures (Td) of collagens were 34.8°C and 35.1°C, respectively. This study shows that there is a possibility to use pipefish collagen as the alternative source of collagen from industrial purposes and subsequently it may evaluate the economical value of the seaweed pipefish.

Protective Effect of GABA-enriched Fermented Sea Tangle against Ethanol-induced Cytotoxicity in HepG2 Cells

Young-Mi Kang,천충길,Bae-Jin Lee,김영목 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.5

This study was conducted to evaluate the protective effects of GABA (gamma-amino butyric acid)-enriched sea tangle juice (STJ) by Lactobacillus brevis BJ-20fermentation against alcohol hepatotoxicity. The protective effects were determined by assessing glutathione (GSH)content levels and gamma-glutamyl transpeptidase (GGT)activity against ethanol-induced cytotoxicity in HepG2cells. In ethanol-treated cells, GSH content decreased to 44.35% of control (ethanol-untreated cell) values; however,treatment with fermented sea tangle juice (FSTJ) at a concentration of 25 μg/mL increased GSH levels to 67.08%. These results suggest that FSTJ may prevent intracellular GSH depletion caused by ethanol consumption. Treatment with FSTJ against alcohol-injured HepG2 cells resulted in a dose-dependent decrease in GGT activity. The expression of cytochrome P450 2E1 (CYP2E1) enzyme, a major contributor to ethanol-induced oxidative stress, was also completely inhibited in FSTJ-treated cells at a concentration of 25 μg/mL. Thus, this study demonstrated that ethanolinduced cytotoxicity could be attenuated by inhibition of GSH depletion, GGT activity, and CYP2E1 expression.