The study was performed to optimize the processing conditions (alkali concentration, extraction time, and temperature) for rockfish skin gelatin based on yield using response surface methodology and comparison of the physicochemical properties with those of rockfish skin gelatin pretreated and extracted under ordinary conditions (alkali treatment concentration: 1.0 M; extraction time: 2 hr; extraction temperature: $60^{\circ}C$). Predicted maximum gelatin yield of 19.1% and gelatin content of 87.8% were obtained by extraction at $106.6^{\circ}C$ for 69.0 min after pretreatment with 1.1 M calcium hydroxide. Yield of gelatin extracted under high temperature/high pressure (G-HT/HP) was 54% higher than that extracted under ordinary temperature/time (G-OT/T). However, G-HT/HP was inferior in gel strength and gelling point to (G-OT/T), but comparable in transmission. Based on the physicochemical properties, G-HT/HP was unsuitable for use in products requiring higher physical properties, but could be useful for health-functional foods.

This study was conducted to evaluate the potential of a microbial fermentation procedure to improve the functional qualities of seaweeds. Aspergillus oryzae, which has been used in traditional Korean fermented foods, was inoculated and cultivated in an aqueous extract of sea tangle (Laminaria japonica). Fermentation of the sea tangle extract by A. oryzae for 4 days resulted in a 3-fold increase in $\gamma$-aminobutyric acid (GABA) content. GABA is known to be a bioactive compound. Fungal fermentation of the extract also enhanced its antioxidant activity and increased its total content of phenolic compounds. It was assumed that these changes stemmed from the biodegradation of active compounds of the sea tangle packaged within its rigid structural matrix or occurred as result of fungal fermentation. These results suggested that the application of microbial fermentation to the processing of seaweeds will help in the development of processed foods to meet consumer demands.

Interest in plant-derived food additives has increased recently, with efforts to identify natural antioxidant sources to replace synthetic antioxidants. This study evaluated the antioxidant effects of organic solvent fractions of 80% methanol extracts from Pediastrum duplex and Dactylococcopsis fascicularis microalgae. Among the solvent fractions tested, the ethyl acetate and n-hexane fractions from P duplex effectively scavenged 79.8% and 74.5% of DPPH free radicals, respectively. The chloroform fraction from D. fascicularis showed the strongest $H_2O_2$ scavenging ability (49.7%). The greatest scavenging of hydroxyl radicals (73.1%) was exhibited by the 80% methanol extract from P duplex. Aqueous residue and ethyl acetate fraction from P duplex provided the strongest nitric oxide scavenging (57.7%) and metal chelating effect (82.1%), respectively. Chloroform and ethyl acetate fractions from P duplex and n-hexane fraction from D. fascicularis exhibited significantly greater inhibition of lipid peroxidation than the commercial antioxidants. These results suggested that P duplex and D. fascicularis microalgae having potential anti oxidative compounds with various properties could be utilized in the food and pharmaceutical industries.

In our previous study, we preliminarily demonstrated that Celluclast and Neutrase extracts exhibited the strongest $H_2O_2$-scavenging activities among five carbohydrases (Viscozyme, Celluclast, Termamyl, Ultraflo and AMG) and five proteases (Kojizyme, Alcalse, Flavourzyme Protamex and Neutrase) extracts. Thus, Celluclast and Neutrase extracts were selected for use in further experiments and were separated into four different molecular weight fractions (<5, 5-10, 10-30 and >30 kDa). Among them, the 5-10 kDa fraction showed the highest $H_2O_2$-scavenging activity. The 5-10 kDa fraction also strongly enhanced cell viability against $H_2O_2$-induced oxidative damage. Furthermore, the fraction reduced the proportion of apoptotic cells induced by $H_2O_2$, as demonstrated by decreased sub-G1 hypodiploid cells and decreased apoptotic body formation by flow cytometry. These results indicated that the 5-10 kDa fraction of the Celluclast and Neutrase extracts from S. coreanum exhibited strong antioxidant activity over $H_2O_2$-mediated cell damage in vitro.

An agar-degrading Agarivorans sp. AG17 strain was isolated from the red seaweed Grateloupia filicina collected from Jeju Island. A beta-agarase gene from Agarivorans sp. AG17 was cloned and designated as agrA. agrA has a 2,985 bp coding region encoding 995 amino acids and was classified into the glycoside hydrolase family (GHF)-50. Predicted molecular mass of the mature protein was 105 kDa. His-tagged agrA was overexpressed in Escherichia coli and purified as a fusion protein. The enzyme showed 158.8 unit/mg specific activity (optimum temperature at $65^{\circ}C$ and pH 5.5 in acetate buffer) with unique biochemical properties (high thermal and pH stabilities). Enzyme produced neoagarohexaose, neoagarotetraose and neoagarobiose by degrading agar, and hydrolyzed neoagaro-oligosaccharides were biologically active. Hence the purified enzyme has potential for use in industrial applications such as the development of cosmetics and pharmaceuticals.

A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.

To determine whether long double-stranded RNA (dsRNA) induces RNA interference and type I interferon (IFN) responses in fish, long dsRNAs encoding enhanced green fluorescent protein (EGFP), GFPuv, and polyinosinic-polycytidylic acid sequences were co-injected with an EGFP expressing plasmid, into rock bream (Oplegnathus fasciatus). We investigated the EGFP mRNA and protein levels, and the transcriptional responses of dsRNA-dependent protein kinase and Mx1 genes. Long dsRNAs were strong inducers of a type I IFN response in rock bream, resulting in nonspecific suppression of exogenous gene expression. Furthermore, sequence-specific knockdown of exogenous gene expression at the mRNA level was detected at an early phase (24 h). These results suggested that long dsRNA may inhibit exogenous gene expression through an early mRNA interference response and a later type I IFN response in fish.

Two feeding experiments were conducted to investigate the effects of dietary genetically modified (GM) soya and com on growth performance, feed utilization and body composition of juvenile olive flounder, Paralichthys olivaceus and rockfish, Sebastes schlegelii. For each fish species, four isonitrogenous (50% crude protein) and isocaloric (4.1 kcal/g) diets (designated as nGM soya, GM soya, nGM com and GM com) were formulated to contain 20% non-GM (nGM) and GM soya and com. Thirty olive flounder (initial body weight, 15.4${\pm}$0.4 g) and fifty rockfish (initial body weight, 3.1${\pm}$0.02 g) were distributed in each 400 L tank (200 L water) in a flow through system. Each experimental diet was fed to triplicate groups of fish to visual satiation, twice a day (9:00 hand 17:00 h) for 6 weeks. Growth performance was measured every three weeks. No effects of GM feedstuffs on survival were observed. Dietary inclusion of GM feedstuffs did not affect growth performance and feed utilization of fishes, except for rockfish fed GM com. Rockfish fed the GM com diet showed higher weight gain, daily feed intake and daily protein intake than did fish fed the nGM com diet, but no significant differences were observed in final body weight between the dietary treatments. Condition factor, hepatosomatic index, visceral somatic index and body composition were not altered by the inclusion of GM feedstuffs. These results indicate that dietary inclusion of GM soya and com could have no effects on growth performance and feed utilization of juvenile flounder and rockfish. Lower weight gain and feed intake in flounder and rockfish fed the diets containing 20% soya were likely due to anti-nutritional factors, rather than transgenic factors in the feedstuffs. Dietary inclusion of GM soya and com at the level tested did not alter the body composition of fishes. Further studies to investigate the effects of GM feedstuffs on health conditions and the development of fishes, as well as those of residue of transgenic fragments in ambient environments and in animals are necessary for safe use of the ingredients in aquaculture.

We investigated the effect of Cu exposure on the activities of protective antioxidant enzymes in the gills and digestive glands of the manila clam Ruditapes phillippinarum exposed to subchronic concentrations (0, 20, 40, and $80{\mu}gL^{-1}$) of waterborne Cu. No mortality occurred during the experimental period, and no significant condition index differences were observed in any exposure group compared with the control. No significant differences were observed in the digestive glands and gills of the clams observed during 15 days of exposure, but after 30 days, the SOD activity in the gill showed a significant difference between the $80{\mu}gL^{-1}$ Cu-exposed group and the control. GPx activities in the digestive glands and gills were significantly lower after 30 days of Cu exposure. Gill GR activity in the high-exposure group ($80{\mu}gL^{-1}$) was significantly elevated compared with that in the control group. GST activities in the digestive glands of all groups did not change over 30 days. However, GST activity in the gill at $80{\mu}gL^{-1}$ Cu was significantly higher after 15 and 30 days of exposure. GSH activities in the gill showed patterns similar to those of GST activities during exposure periods. In the digestive glands, GSH activity was higher only at $80{\mu}gL^{-1}$ after 30 days exposure. In digestive glands and gills, the MDA levels of clams exposed to $80{\mu}gL^{-1}$ Cu were significantly higher after 30 days of exposure.

Since 1998, a new viral disease with high mortality has been consistently recorded in Korea in cultured carp, Cyprinus carpio. In this study, we investigated an epizootic of the disease that caused high mortality rates in carp obtained from 11 farms in Korea between 1999 and 2007. Assessment of koi herpesvirus (KHV) levels in diseased carp was carried out to determine if this virus was the etiologic agent of disease in this instance. High mortality rates in carp were recorded mainly in the spring and autumn at water temperatures between $19^{\circ}C$ and $24^{\circ}C$. Diseased fish typically showed surface discoloration, with a thick opaque mucus covering the body and gills. Protozoan parasites and bacteria were recovered from 7/29 (24%) and 2/26 (8%) of fish, respectively. Evidence of viral infection was marked; cytopathic effects (CPEs), characterized by cell rounding and an extended cytoplasm in fathead minnow (FHM) cells, were detected in 40/41 fish (98%). A high mortality rate (80%) resulted when supernatants of cell cultures showing CPEs were applied to previously healthy fish. KHV was detected by polymerase chain reaction in 6/41 fish (15%), but was not detected in supernatants obtained from cell cultures showing CPEs. These results suggest that KHV may not be the etiologic agent of the high mortality occurring among cultured carp in Korea; therefore, some other-as yet unidentified-infective agent must be responsible.

A peptide that inhibits HIV-1 protease was isolated from a hydrolysate of oyster (Crassostrea gigas) proteins digested with thermolysin. The peptide was using membrane filtration, gel permeation chromatography, ion exchange chromatography, and reverse-phase high performance liquid chromatography. Amino acid sequence of the peptide was determined to be Val-Phe-Glu-Leu. Chemically synthesized Val-Phe-Glu-Leu showed an $IC_{50}$ value of 106 ${\mu}M$.