• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2000.11a
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• pp.59-75
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• 2000

This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.3
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• pp.260-267
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• 2001

The biochemical composition and antioxidative activity of marine microalgae were investigated for the effective utilization of marine resources. Two species of marine microalgae, Nannochloris oculata (N. oculata) of Chlorophyceae and Phaeodactylum tricornutum (P. tricornutum) of Bacillariophyceae, were selected. Because these species showed the high growth rate and easy to continuous culture. The contents of crude protein, lipid, and carbohydrate were $54.91\%,\;11.29\%,\;and\;10.15\%$, for N. oculata and $38.07\%,\;13.19\%,\;and\;7.13\%$, for P. tricornutum, respectively. Glutamic acid was the highest concentration for both species. Galactose (3,712.02 mg/100g), fucose (1,966.03 mg/100g), and glucose (1,814.35 mg/100g) were the major carbohydrates for N. oculatae, and glucose (5,295.45 mg/100g) and mannose (841.34 mg/100g) were for P. tricornutum. K (12,906.86 mg/100g), Mg (1,039.15 mg/100g), Ca (882.57 mg/100g) and Fe (747.20 mg/100g) were the major minerals for N. oculata, and K (11,718.65 mg/100g), Ca (2,003.32 mg/100g), Mg (1,570.84 mg/100g) and Fe (552.58 mg/100g) were for P. tricornutum. In the composition of nucleotides, ADP ($4.77{\mu}mol/g$) was the highest in N. oculata and hypoxanthine (11.74{\mu}mol/g) in P. tricornutum. Large amount of linoleic acid (18: 2, $\omega-6$) was contained in N. oculata. In contrast 16: 1 ($\omega-7$) and 20: 5 ($\omega-3$) were major fatty acid in P. tricornutum. The antioxidative activities of organic solvent extracts of two microalgae were measured by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay method. The chloroform extract obtained from P. tricornutum was identified to be the most effective in DPPH radical scavenging activity.

• Microbiology and Biotechnology Letters
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• v.33 no.4
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• pp.274-280
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• 2005

Pasteurella multocida is known to cause widespread infections in husbandry. To induce homologous and heterologous immunity against the infections, outer membrane proteins (OMPs) in the envelope of P. multocida are thought to be attractive vaccine candidates. Outer membrane protein H is considered as the major component of OMPs. In this study, a gene for OmpH was isolated from pathogenic P. multocida serogroup A. The gene was composed of 1,047 nucleotides coding 348 amino acids with signal peptide of 20 amino acids. The amino acid composition showed about 80 to 98 per cent sequence homologies among other 10 strains of P. multocida serogroup A, reported so far. A recombinant ompH, from which signal peptide was truncated, was generated using pRSET A to name 'pRSET A/OmpH-F2'. The pRSET A/OmpH-F2 was well expressed in E. coli BL21(DE3). The truncated OmpH was purified using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Its molecular weight was registered to be 40 kDa on SDS-PAGE gel. In order to generate immunesera against the OmpH, 50 ug of the protein was intraperitoneally injected into mice three times. The anti-OmpH immuneserum recognized about $5{\times}10^{-2}$ng quantity of the purified OmpH. It can be used for an effective vaccine production to prevent fowl cholera caused by pathogenic P. multocida (Serogroup A).

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.993-999
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• 2010

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

• Journal of Aquaculture
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• v.11 no.3
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• pp.319-325
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• 1998

Quality evaluation of fish muscle was conducted in olive flounder, Paralichthys olivaceus fed with control or experimental diet containing 0.3% of Obosan (Sungam Co., Korea). Sensory panel members preferred the experimental fish were more preferable than those of control (P<0.01 or 0.05). The difference test was shown that the experimental fish muscle is more firm elastic and palatable than the control ones (P<0.01 or 0.05). The physical properties of meats from fish fed a diet containing Obosan improved qualities with compared to those of control meats. The total amount of free amino acids in muscles from control fish (31.7 mg/100mg), especially with the significant increase of glutamic acid, proline, glycine, alanine and methionine. In all nucleotides and their related compounds, ATP was the most abundant in muscles from fish fed a diet containing Obosan (about 1.9 fold compared to mean of control), however, IMP was more abundant in control muscles.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.02a
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• pp.108-109
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• 2013

Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.1
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• pp.41-51
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• 1987

This study was carried out to prepare powdered smoked-dried mackerel which can be used as a soup base, and to examine storage stability and the taste compounds of Products. Raw mackerel are filleted, toiled for 10 minutes and pressed to remove lipids, and then soaked in extract solution of skipjack meat. This soaked mackerel are smoked 3 times to $10-12\%$ moisture content at $80^{\circ}C$ for 8 hours. And the smoked-dried mackerel were pulverized to 50 mesh. Finally, the powdered smoked-dried mackerel were packed in a laminated film $bag(PET/Al\;foil/CPP:\;5{\mu}m/15{\mu}m/70{\mu}m,\;15\times17cm)$ with air(product C), nitrogen(product N) and oxygen absorber(product O), and then stored at room temperature for 100 days. The moisture and crude lipid content of powdered smoked-dried mackerel was $11.3-12.3\%,\;12\%$, respectively, and water activity is 0.52-0.56. And these values showed little changes during storage. The pH, VBN and amino nitrogen content increased slowly during storage. Hydrophilic and lipophilic brown pigment formation showed a tendency of increase in product(C) and showed little change in product(N) and (O). The TBA value, peroxide value and carbonyl value of product(N) and (O) were lower than those of product (C). The major fatty acids of products were 16:0, 18:1, 22:6, 18:0 and 20:5, and polyenoic acids decreased, while saturated and monoenoic acids increased during processing and storage of products. The IMP content in products were 420.2-454.2 mg/100 g and decreased slightly with storage period. And major non-volatile organic acids in products were lactic acid, succinic acid and $\alpha-ketoglutaric$ acid. In free amino acids and related compounds, major ones are histidine, alanine, hydroxyproline, lysine, glutamic acid and anserine, which occupied $80.8\%$ of total free amino acids. The taste compounds of powdered smoked-dried mackerel were free amino acids and related compounds (1,279.4 mg/100 g), non-volatile organic acids(948.1 mg/100 g), nucleotides and their related compounds (672.8 mg/100 g), total creatinine(430.4 ntg/100 g), tetaine(86.6 mg/100 g) and small amount of TMAO. The extraction condition of powdered smoked-dried mackerel in preparing soup stock is appropriate at $100^{\circ}C$ for 1 minute. Judging from the results of taste and sensory evaluation, it is concluded that the powdered smoked-dried mackerel can be used as natural flavoring substance in preparing soups and broth.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.14 no.4
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• pp.201-211
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• 1981

For the effective utilization of sardine, sardinops melanosticta, one of the major coastal fish in Korea, of which annual catch has been increasing from year to year since 1970, it was processed in form of fermented fish paste. The fish were treated with BHA and Teaox-Ⅱ in concentration of $0.01\%\;and\;0.02\%$ to prevent the oxidation of lipid during fermentation and then salted with $20\%$ table salt and fermented at room temperature of $25\pm3^{\circ}C$. The duration of fermentation necessary for the final product with an acceptable taste was determined by sensory evaluation by means of profile method. From the result of sensory evaluation, one month was found to be suitable as the reasonable duration of fermentation. Both BHA and Tenox-Ⅱ in conceatration of $0.02\%$showed a good preventing effect on the lipid oxidation during fermentation. In case of fermented sardine treated with both antioxidants, lipid oxidation occurred little up to two months, whereas the control showed a remarkable deterioration during one month of fermentation. Most of the nucleotides in sardine was decomposed from adenosing triphosphate to inosine and hypoxanthine during the fermentation of one month. The great portion of free amino acids in the extractives of product was occupied by leucine, glutamic acid, isoleucine, alanine, valine and ysine in turn, and their coatent was $59.4\%$ of the total free amino acids. The amount of essential amino acids was $59.4\%$ of the total free amino acids. The contents of 5'-IMP, betaine, trimethylamine oxide and total creatinine in the extractives of product were $1.9{\mu}mole/g,\;4.9mg\%,\;1.0mg\%\;and\;475mg\%$, respectively. According to the omission test, the main constituents of the characteristic taste of fermented sardine could be assumed as free amino acids and a little amount of 5'-IMP.

• Journal of the Korean Society of Food Science and Nutrition
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• v.9 no.1
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• pp.23-32
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• 1980

This study was attempted to establish the basic data for evaluating taste compounds in fermented entrails of Clupanodon Osdeckii. The changes of such compounds as amino acids, nucleotides and their related compounds, betaine, TMAO and TMA during fermentation were analyzed. IMP, AMP, ADP and ATP were decreased, while hypoxanthine was increased during the fermentation. The content of hypoxanthine in fermented entrails of Clupanodon Osdeckii after 50 days was increased to about 2 times of that in raw entrails. In the free amino acid composition of raw entrails, abundant amino acids were lysine, glutamic acid, valine, alanine, threonine, serine, leucine and glycine in order. Such amino acids as arginine, tyrosine and phenylalanine were lower than 2.0% of total free amino acid, and proline and cysteine were detected in trace amount. The changes in free amino acid composition of the extract in entrails of Clupanodon Osdeckii during fermentation were not observed. Such amino acids as lysine, glutamic acid, valine, serine and leucine were especially abundant in both raw and fermented products. The content of total free amino acids in fermented entrails of Clupanodon Osdeckii after 50 days were increased to about 12 times of that in raw. The content of betaine nitrogen were about 14.5 (moisture and salt free base) after 50 days of fermentation. TMAO nitrogen was decreased during the fermentation. It is believed that lysine, glutamic acid, valine, serine, leucine and hypoxanthine play an important role as taste compounds in fermented entrails of Clupanodon Oseckii.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.99-104
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• 2009

The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.