• Title/Summary/Keyword: LEU

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Effect of Selected Inoculant Applications on Chemical Compositions and Fermentation Characteristics of High Moisture Rye Silage

  • Lee, Seong Shin;Jeong, Seung Min;Seo, Myeong Ji;Joo, Young Ho;Paradhipta, Dimas Hand Vidya;Seong, Pil Nam;Kim, Sam Churl
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.42 no.3
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    • pp.155-161
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    • 2022
  • The aim of this study was to investigate the effect of isolated lactic acid bacteria (LAB) on the quality of high moisture rye silage. Rye forage (Secale cereale L.) was harvested at the heading stage (27.3% of dry matter (DM)) and cut into approximately 3-5 cm lengths. Then, the forage divided into 4 treatments with different inoculants: 1) No additives (CON); 2) Lactobacillus brevis strain 100D8 at a 1.2 × 105 colony-forming unit (cfu)/g of fresh forage (LBR); 3) Leuconostoc holzapfelii strain 5H4 at a 1.0 × 105 cfu/g of fresh forage (LHO); and 4) Mixture of LBR and LHO (1:1 ratio) applied at a 1.0 × 105 cfu/g of fresh forage (MIX). About 3 kg of forage from each treatment was ensiled into a 20 L mini-bucket silo in quadruplicate for 100 days. After silo opening, silage was collected for analyses of chemical compositions, in vitro nutrient digestibilities, fermentation characteristics, and microbial enumerations. The CON silage had the highest concentrations of neutral detergent fiber and acid detergent fiber (p = 0.006; p = 0.008) and a lowest in vitro DM digestibility (p < 0.001). The pH was highest in CON silage, while lowest in LBR and MIX silages (p < 0.001). The concentrations of ammonia-N, lactate, and acetate were highest in LBR silage (p = 0.008; p < 0.001; p < 0.001). Propionate and butyrate concentrations were highest in CON silage (p = 0.004; p < 0.001). The LAB and yeast counts were higher in CON and LHO silages compare to LBR and MIX silages (p < 0.001). However, the mold did not detect in all treatments. Therefore, this study could conclude that L. brevis 100D8 and Leu. holzapfelii strain 5H4 can improve the digestibility and anti-fungal activity of high moisture rye silage.

Food Characteristics of Protein Isolates Recovered from Olive Flounder Paralichthys olivaceus Roe by Isoelectric Solubilization and Precipitation Process (넙치(Paralichthys olivaceus) 알로부터 등전점 용해/침전공정에 의해 회수한 분리단백질의 식품특성)

  • Sang in Kang;In Sang Kwon;Hyeung Jun Kim;In Seong Yoon;Yu Ri Choe;Jung Suck Lee;Jin-Soo Kim;Min Soo Heu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.2
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    • pp.162-173
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    • 2023
  • Four roe protein isolates (RPIs) from olive flounder Paralichthys olivaceus roes (OFR) were recovered by isoelectric solubilization (pH 11 and 12) and precipitation (pH 4.5 and 5.5) and investigated for their food characteristics. RPIs contained 4.5-9.6% moisture, 64.1-69.5% protein, 16.1-19.8% lipid, and 1.0-3.9% ash. The protein yields of RPIs ranged from 50.1 to 56.8%, which was significantly different depending on the recovery conditions. A difference was observed in the SDS-PAGE protein band (25-100 kDa) between the alkaline solubilization at pH 11 (RPI-1 and 2) and pH 12 (RPI-3 and 4). The major amino acids of RPIs were Leu, Lys, Asp, Glu and Ala and major mineral components were sulfur, sodium, phosphorus, and magnesium, which were significantly different from OFR (P<0.05). Additionally, the lead and cadmium content was below the chemical hazard standard of the Korean food standards code. The Hunter color and whiteness of RPIs also showed significant differences according to the treatment conditions of the ISP process (P<0.05). This suggests that protein isolates recovered from olive flounder roes have high potential as nutritional supplements.

Cloning and Functional Studies of Pro-Apoptotic MCL-1ES BH3M (세포사멸을 유도하는 새로운 단백질인 MCL-1ES BH3M의 클로닝 및 기능연구)

  • Kim, Jae-Hong;Park, Mira;Ha, Hye-Jeong;Lee, Kangseok;Bae, Jeehyeon
    • Development and Reproduction
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    • v.12 no.3
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    • pp.297-303
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    • 2008
  • BCL-2 family members are essential protein for the regulation of cell death and survival consisting both antiapoptotic and pro-apoptotic proteins. In the present study, we designed and cloned a new apoptotic molecule MCL-1ES BH3M coding a modified protein of MCL-1L. Compared to MCL-1L protein, MCL-1ES BH3M lacks the PEST motifs known to be involved in MCL-1L protein degradation and has seven mutated residues in BH3 domain critical for dimerization with BCL-2 family members. Overexpression of MCL-1ES BH3M induced death of different cells, and its cell killing effect was not blocked by forced expression of the pro-survival protein MCL-1L. Expression of MCL-1ES BH3M protein led to the activation of caspase 9 and caspase 3, suggesting apoptotic cell death, and confocal fluorescent microscopic analyses showed that MCL-1ES BH3M was partially localized in mitochondria. In conclusion, we reported a new apoptotic molecule and determined its cell death activity in cells.

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Comparison of amino acid digestibility of soybean meal, cottonseed meal, and low-gossypol cottonseed meal between broilers and laying hens

  • Kai Qiu;Xiao-cui Wang;Jing Wang;Hao Wang;Guang-hai Qi;Hai-jun Zhang;Shu-geng Wu
    • Animal Bioscience
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    • v.36 no.4
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    • pp.619-628
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    • 2023
  • Objective: This study aimed to determine and compare the apparent ileal digestibility (AID) and the standardized ileal digestibility (SID) of amino acids (AA) in soybean meal (SBM), cottonseed meal (CSM), and low-gossypol cottonseed meal (LCSM) fed to broiler chickens and laying hens. Methods: Three semi-purified diets containing the identical crude protein concentration at 20% were formulated to contain SBM, CSM, or LCSM as the sole source of N. A N-free diet was also formulated to estimate the basal ileal endogenous losses of AA for broilers and hens. A total of 300 male Ross 308 chicks at one-day-old and 144 Hy-Line Brown laying hens at 30-week-old with initial egg production rate of 88.3%±1.0% were randomly allocated into 1 of 4 dietary treatments, respectively. Results: CSM and LCSM showed more Arg and Cys+Met while less Lys, Ile, Leu, and Thr relative to SBM. Significant interactions existed between species and experimental diets for AID (except for Arg, Asp, Glu, Gly, and Pro) and SID (except for Arg, His, and Phe) of most AA. Most AA in diets showed higher AID (except for Lys) and SID (except for Lys, Met, and Ser) in broilers relative to laying hens. The AID and SID of all AA were significantly different between the three diets. In broilers, the AID and SID of most indispensable AA except for Arg in SBM and LCSM was higher than CSM. In laying hens, the AID and SID of most indispensable AA except for Arg, Met+Cys, and Phe in SBM was higher than CSM and LCSM. Conclusion: The accurate determination of AID and SID of AA in CSM and LCSM for broilers and layers benefits the application of CSM and LCSM in chicken diets. The cottonseed by-products CSM or LCSM showed the species-specific AA digestibility values for broilers and layers.

Recombinant human KAI1/CD82 attenuates M1 macrophage polarization on LPS-stimulated RAW264.7 cells via blocking TLR4/JNK/NF-κB signal pathway

  • Hyesook Lee;Jung-Hwa Han;Kangbin An;Yun Jeong Kang;Hyun Hwangbo;Ji Hye Heo;Byung Hyun Choi;Jae-Joon Kim;Seo Rin Kim;Soo Yong Lee;Jin Hur
    • BMB Reports
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    • v.56 no.6
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    • pp.359-364
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    • 2023
  • KAI1/CD82, a membrane tetraspanin protein, can prevent various cancers and retinal disorders through its anti-angiogenic and anti-metastatic capacity. However, little is known about its anti-inflammatory effect and molecular mechanism. Therefore, the present study aimed to inLPSvestigate effect of a recombinant protein of the large extracellular domain of human KAI1 (Gly 111-Leu 228, rhKAI1) on lipopolysaccharides (LPS)-stimulated RAW264.7 macrophage-like cells and mouse bone marrow-derived macrophages (BMDM) and to identify its underlying mechanism. Our data showed that rhKAI1 suppressed expression levels of classically macrophages (M1) phenotype-related surface markers F4/80+CD86+ in LPS-stimulated BMDM and RAW264.7 cells. In addition, LPS markedly increased mRNA expression and release levels of pro-inflammatory cytokines and mediators such as interleukin (IL)-1β, IL-6, tumor necrosis factor-α, cyclooxygenase-2, nitric oxide and prostaglandin E2, whereas these increases were substantially down-regulated by rhKAI1. Furthermore, LPS strongly increased expression of NF-κB p65 in the nuclei and phosphorylation of ERK, JNK, and p38 MAPK. However, nuclear translocation of NF-κB p65 and phosphorylation of JNK were greatly reversed in the presence of rhKAI1. Especially, rhKAI1 markedly suppressed expression of toll-like receptor (TLR4) and prevented binding of LPS with TLR4 through molecular docking predict analysis. Importantly, Glu 214 of rhKAI1 residue strongly interacted with Lys 360 of TLR4 residue, with a binding distance of 2.9 Å. Taken together, these findings suggest that rhKAI1 has an anti-inflammatory effect on LPS-polarized macrophages by interacting with TLR4 and down-regulating the JNK/NF-κB signaling pathway.

Application of peak based-Bayesian statistical method for isotope identification and categorization of depleted, natural and low enriched uranium measured by LaBr3:Ce scintillation detector

  • Haluk Yucel;Selin Saatci Tuzuner;Charles Massey
    • Nuclear Engineering and Technology
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    • v.55 no.10
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    • pp.3913-3923
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    • 2023
  • Todays, medium energy resolution detectors are preferably used in radioisotope identification devices(RID) in nuclear and radioactive material categorization. However, there is still a need to develop or enhance « automated identifiers » for the useful RID algorithms. To decide whether any material is SNM or NORM, a key parameter is the better energy resolution of the detector. Although masking, shielding and gain shift/stabilization and other affecting parameters on site are also important for successful operations, the suitability of the RID algorithm is also a critical point to enhance the identification reliability while extracting the features from the spectral analysis. In this study, a RID algorithm based on Bayesian statistical method has been modified for medium energy resolution detectors and applied to the uranium gamma-ray spectra taken by a LaBr3:Ce detector. The present Bayesian RID algorithm covers up to 2000 keV energy range. It uses the peak centroids, the peak areas from the measured gamma-ray spectra. The extraction features are derived from the peak-based Bayesian classifiers to estimate a posterior probability for each isotope in the ANSI library. The program operations were tested under a MATLAB platform. The present peak based Bayesian RID algorithm was validated by using single isotopes(241Am, 57Co, 137Cs, 54Mn, 60Co), and then applied to five standard nuclear materials(0.32-4.51% at.235U), as well as natural U- and Th-ores. The ID performance of the RID algorithm was quantified in terms of F-score for each isotope. The posterior probability is calculated to be 54.5-74.4% for 238U and 4.7-10.5% for 235U in EC-NRM171 uranium materials. For the case of the more complex gamma-ray spectra from CRMs, the total scoring (ST) method was preferred for its ID performance evaluation. It was shown that the present peak based Bayesian RID algorithm can be applied to identify 235U and 238U isotopes in LEU or natural U-Th samples if a medium energy resolution detector is was in the measurements.

The Growth Characteristics and Germanium Uptake by Water Celery in Soil Treated with Germanium (게르마늄 처리 토양에서 미나리 생육 특성과 게르마늄 흡수)

  • Lee, Seong-Tae;Lee, Young-Han;Heo, Jae-Young;Hong, Kwang-Pyo;Dahlgren, Randy A.;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.27 no.2
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    • pp.185-190
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    • 2008
  • In order to obtain the basic information for agricultural utilization of Germanium(Ge), the growth characteristics and the germanium uptake by water celery were investigated at different concentration of germanium in soil. This experiment was carried out in the Wagner pot(1 $5,000^{-1}a$). Germanium concentrations in soil for water celery cultivation were maintained at 0.26, 25.0, 62.5, and 125.0 mg $kg^{-1}$, respectively. The treatment of over Ge 25.0 mg $kg^{-1}$ in the soil led to germanium phytotoxicity such as reduction of plant height and fresh weight. The contents of germanium in water celery were increased with the increase of germanium concentration in the soil. When water celery was cultivated from soil maintained with Ge 25.0 and 62.5 mg $kg^{-1}$, its germanium contents in plant were 89.9 and 371.6 mg $kg^{-1}$, respectively. Then, the efficiency of germanium uptake of water celery in Ge 25.0 and 62.5 mg $kg^{-1}$ maintained plots was 1.7 and 2.4%, respectively. When water celery was cultivated from soil maintained with Ge 25.0, 62.5 and 125.0 mg $kg^{-1}$, its content of amino acid was found to be 89.8, 198.4, and 318.2 mg $g^{-1}$, respectively. To investigate the effect of N fertilizer application in uptake of germanium by water celery, these were treated with nontreatment, 1.0, 1.5 and 2.0 times of N application based on soil testing for cultivation of water celery. However, the amount of the N fertilizer application did not affect the contents of germanium in the water celery. When water celery was cultivated from soil maintained with two kinds of inorganic and organic germanium 50 mg $kg^{-1}$, respectively, the content of germanium were 24.2 mg $kg^{-1}$ in the Ge-132 treatment and 11.8 mg $kg^{-1}$ in the $GeO_2$ treatment.

Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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Properties of the High and Low Molecule of the Proteoglycan Extracted from Ganoderma lucidum IY009 (Ganoderma lucidum IY009 배양균사체 유래 단백다당류의 저분자와 고분자 분획의 특성)

  • Baek, Seong-Jin;Kim, Yong-Seuk;Chun, Uck-Han;Lee, Eun-Sook;Lee, June-Woo
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.1-8
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    • 2001
  • To examine the structural properties of the proteoglycan (GMPG, Ganoderma lucidum mycelial proteoglycan) obtained from mycelia in Ganoderma lucidum IY009, we obtained the low and high molecular proteoglycan by ultrafiltration and sepharose CL-4B column chromatography. The physicochemical properties of these fractions were as follows. When the proteoglycan separated by ultrafiltration and sepharose CL-4B column chromatography, its was not fractionated completely. The molecular weight of high molecular proteoglycan by the gel column chromatography (CH) was 250 kD and 2,000 kD, and low molecular proteoglycan was 12kD. The total carbohydrate was consisted of 75.7% (UH) and 96.7% (CH), and the low fraction was 72.7% (UL) and 87.1% (CL), respectively. The sugar of high and low molecular proteoglycan composed of glucose, mannose, fructose, galactose, xylose, ribose and arabinose. Glucose contents of all fraction were ranged from $46.9%{\sim}82.4%$ of the total sugar and the ratio of ${\alpha}$\;and\;{\beta}-glucose$ was $0.84{\sim}1.14$, and its indicated the proteoglycan to be ${\beta}-glucan$. Amino acids pattern showed that the fractions contained a large amount of aspartie acid, glutamic acid, alanine and leucine. These fractions showed the characteristics of IR absorption for ${\beta}-glucan$ at $890\;cm^{-1}\;and\;^{13}C-NMR$ spectroscopy showed the presence of the ${\beta}-1,3-glucan$ and a ${\beta}-1,6-glucan$.

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Quality Characteristics of the Salt-Fermented Oysters in Olive Oil (기름담금 염장발효 굴의 품질특성)

  • Kim, Seok-Moo;Kong, Chung-Sik;Kim, Jong-Tae;Kang, Jeong-Koo;Kim, Nam-Woo;Kim, Jeong-Bae;Oh, Kwang-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.8
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    • pp.1398-1406
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    • 2004
  • To develop the new type of salt-fermented seafoods, the salt-fermented oysters in olive oil (product SO) were manufactured, and food components and quality characteristics of product SO were examined. The optimum processing condition for product SO is as follows. The raw oyster with no shell was washed off with 3% saline solution. Then dewatered, and dipped in the brine-salting solution made up with saturated saline solution and oyster sauce (2 : 1 v/v) mixture added 1% sodium erythorbic acid and 0.2% polyphosphate. After salt-fermentation it ripened by brine salting at 5$\pm$1$^{\circ}C$ for 15 days. Then dried at 15$^{\circ}C$ for 4 hours with cool-air, and packed in No. 3B hexahedron type can. Finally, poured with olive oil and seamed it by double-seamer. The moisture, crude protein, crude ash and volatile basic nitrogen contents of the product SO were 61.6%, 12.0%, 16.3% and 34.3 mg/100 g, respectively. In taste-active components of the product SO, total amount of free amino acids is 2,335.4 mg/100 g and it has increased by 50% overall during salt-fermentation 15 day. Taurine, glutamic acid, proline, glycine, alanine, $\beta$-alanine and lysine were detected as principal free amino acids. The contents of inorganic ions were rich in Na and K ion, while the amounts of nucleotide and its related compounds and other bases except betaine were small. From the results of this research, the product SO had a superior organoleptic qualities compared with conventional oyster product, and could be reserved in good conditions for storage 90 days at room temperature.