• Korean Journal of Food Science and Technology
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• v.17 no.5
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• pp.389-398
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• 1985

To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.332-338
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• 2009

A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.

• Journal of Dairy Science and Biotechnology
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• v.14 no.2
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• pp.157-164
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• 1996

This study aimed increase the quality during ripening of Cheddar cheese made with proteinase-negative mutant of Streptococcus lactis KCTC 1913 selected by curing. The degradation of protein during cheese ripening were investigated by electrophoresis and chromatography. The results were summarized as follow ; 1. The number of lactic acid bacteria decreased with the ripening stage, and that of the control cheese decreased faster than that of the cheese made with mutant. 2. Polyacrylamide gel electrophoretic analysis of cheese caseins revealed no difference between the cheese made with mutant and the control cheese, but differences along with the ripening stage were evident. 3. On Sephadex G-25 column chromatography, the extracts of bitter components from the green cheese and 3 month ripended cheese were fractionated into 3 fractions. With the progress of ripening, bitter peptides were degraded to rather small peptides or free amino acids. 4. Sensory evaluation of the 3 month ripended Cheddar cheese found no significant differences in color but the cheese made with mutant evidenced higher palatability in flavor and better texture than the control cheese. 5. The yields of the cheddar cheese made with mutant was 0.14% higher than that of the control cheese.

• Journal of Sericultural and Entomological Science
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• v.28 no.1
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• pp.1-14
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• 1986

This study was carried out to acguire some basic information on nutritional and lhysiological effects of vesicular arbuscular mycorrhizae(VAM) on mulberry trees inoculated with Glomus mosseae, Gerd. & Trap. grown in clay for 65 days and treated with 5 different levels of phosphorus, ie 30, 60, 120, 240, 480 ppm as (NH4)2 HPO4. At the End of the expermental period the levels of fixed phosphate in the soil was measared. And the native VAM fungi were collected to select the most effective VAM species on mulberry tree. The nutritional and biochemical effects of VA mycorrhizae on the mulberry leaves were also studied. Those results are as follow. 1. The mulberry trees grown in clay and inoculated with VAM were heavier in shoot dry weight as much as 197% than uninoculated plants. But in vermioulite, uninoculated mulberry trees were heavier as much as 135% than inoculated. 2. The rates of endo mycorrhizal formation in clay was highest at 60 ppm level of phosphorus, and vesicles in roots were formed in 240ppm and 480ppm level of phosphorus, but not in 30ppm, 60ppm and 120ppm. 3. The greatest growth responses of VAM inoculation was found at 60ppm level of hosphorus, and the optimum phosphorus level for VAM responese appeared to be 60ppm. 4. VAM was also to absorb soil-fixed phosphate. VAM abosrbed Fe-bound phosphate most efficiently and Ca-bound phosphate with ease but not Al-bound phosphate and Al-Fe occluded phosphate. 5. Three species of Gigaspora and one species of Glamus growing naturally in mulberry plantations were collected and tested for the growth responses. Gigaspora tricalipta and Gigaspora calospora revealed the greatest growth responses on mulberry tree among tested VAM fungi. 6. Mulberry leaves inoculated with VA mycorrhizal fungi contained 9.8% more phosphate and 15.2% more nitrogen, 22.2% more water-soluble carbohydrates and 15.2% more proteins than uninoculated plants. 7. The electrophoretic pattern of mulberry leaf protein inoculated with VAM fungi has 19 bands. 8. The patterns of peroxidase zymogram and Amulase zymogram were different between the mulberry leaves inoculated and uninoculated with VA mycorrhizal fungi. The peroxidase zymogram of inoculated leaves has 1 less majour band than unioculated leaves The amylase zymogram of inoculated leaves has 2 bands near the +pole, but that of uninoculated leaves has 1 band near the $pole.

• Tuberculosis and Respiratory Diseases
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• v.53 no.5
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• pp.519-529
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• 2002

Background : The therapeutic effects of surfactants on acute lung injury derive not only from their recruiting action on collapsed alveoli but also from their anti-inflammatory action in the alveolar sapce. This study evaluated the anti-inflammatory action of a surfactant in an acute lung injury model of rats by neutrophils were recollected from the BAL fluid and the NF-${\kappa}B$ activity of the neutrophilic nuclear protein was evaluated. Methods : Male Sprague-Dawley rats weighing approximately 300 gram were divided into 3 groups, which consisted of 6 rats respectively. In the control group, normal saline(3ml/kg) was instilled into the trachea twice with 30 minute interval. In two other groups, acute lung injury was induced by the intra-tracheal instillation of LPS(5mg/kg). Thirty minutes later, either a surfactant(ST group; 30mg/kg) or normal saline(NT group: 3ml/kg) was instilled via the trachea. Twenty-four hours after the LPS instillation, the BAL fluid was retrieved to measure the WBC count and cytokine(IL-$1{\beta}$ and IL-6) levels. The neutrophils were isolated from the BAL fluid and the nuclear protein was extracted to evaluate the NF-${\kappa}B$ activity using a eletrophoretic mobility shift assay(EMSA). Results : The WBC count of the BAL fluid of the ST group($3,221{\pm}1,914{\times}10^3/{\mu}l$) was higher than that of the control group($356{\pm}275{\times}10^3/{\mu}l$)(p<0.05) and lower than that of the NT group($5,561{\pm}1,757{\times}10^3/{\mu}l$)(p<0.05)). The BAL fluid level of IL-$1{\beta}$ from the NT group($2,064{\pm}1,082pg/ml$) was higher than those of the ST group($360{\pm}234pg/ml$)(p<0.05) and the control group(0pg/ml)p<0.05) and control group($49{\pm}62pg/ml$)(p<0.05). The NF-${\kappa}B$ activity of the neutrophilic nuclear protein in the ST group and NT group was similar. Conclusion : The surfactant, attenuates the alveolar inflammation in the acute lung injury of rats model. However, its anti-inflammatory action does no't appear to be mediated by the inhibition of NF-${\kappa}B$ activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.15 no.2
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• pp.123-136
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• 1982

We investigated the changes in protein and free amino acid compositions of the muscles, and amino acid composition of the muscle proteins during postmortem storage of dorsal white and lateral dark muscles of Yellowtail, Seriola quinqueradita, which were kept at $2^{\circ}C$. We present an extensive discussion on the relationship between the changes of freshness and those of protein compositions in the white and the dark muscle of the red-fleshed fish by analyzing polyacrylamide gel electrophoretograms of $NaDodSO_4-solubilized$ sarcoplasmic and myofibrillar proteins extracted from the both muscles. By assessing K-value, total volatile basic nitrogen and pH value as a criterion of freshness, we found that the dark muscle undergoes a more rapid decrease in its freshness compared to that of the white muscle. The contents of the sarcoplasmic and the myofibrillar protein were decreased with postmortem aging of the muscles while those of the residual intracellular protein were increased, and these changes were somewhat faster in the dark muscle than in the white muscle. From the analysis of the electrophoretograms and their densitograms, we found that the sarcoplasmic proteins of the white and the dark muscle were respectively composed of 16 and 12 components. The sarcoplasmic protein of the white muscle lapsed for 10 days showed an increase of 18,000 and 41,000 dalton components, and a gradual decrease of 23,000 and 23,500 dalton components, whereas the sarcoplasmic protein of the dark muscle lapsed for 9 days showed a decrease of 49,000 dalton component, an appearence of a newly formed component of 47,000 dalton, and a disappearance of 26,000 dalton component. The electrophoretograms of the myofibrillar proteins shelved that the white and the dark muscle were composed of 17 and 16 components, respectively. Depending on the lapsed time of postmortem under the controlled condition, the myofibrillar proteins of the white muscle showed an increase of 40,000 dalton component, a gradual decrease of 37,500 dalton component, an appearance of a newly forming component of 32,000 dalton and a disappearance of 26,000 dalton component. On the other hand, the myofibrillar proteins of the dark muscle showed an increase of 58,000 and 64,000 dalton bands, a disappearance of light chain-2 protein and an appearance of a newly forming protein of 32,000 dalton. These changes on the electrophoretic patterns in the dark muscle were more rapid than those in the white muscle. In almost all of the cases, we observed that the changes in the sarcoplasmic protein were faster than those in the myofibrillar protein. The analysis of amino acid of the both muscle proteins showed that the white muscle was rich in glutamic acid, aspartic acid, leucine, arginine, lysine, etc. but was poor in proline and tryptophan. No significant difference was found in the amino acid composition of protein of both the white and the dark muscles. The sample of white muscle lapsed for 10 days shows a remarkable decrease in glutamic and aspartic acids, while that of the dark muscle lapsed for 9 days shows an appreciable decrease in alanine, glycine and arginine. The free amino acid compositions of the white and the dark muscles are respectively characterized with $63\%$ of histidine and $67\%$ of taurine with respect to the total free amino acids of the yellowtail at-death, respectively. The white muscle lapsed for 10 days showed an increase of histidine, valine and taurine, and a slight decrease of alanine, leucine and glycine. The dark muscle lapsed for 9 days shelved an increase of taurine, phenylalanine and glycine, and a decrease of histidine, alanine and serine.

• Korean Journal of Agricultural Science
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• v.16 no.2
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• pp.179-200
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• 1989

Mucor miehei rennet(MR) was added as calf rennet(CR) substitutes in the fixed amounts of mixed rennets in making Camembert cheese. The conditions in the variations of chemical composition: water-soluble nitrogen, non-caseinic nitrogen, non-proteinic nitrogen, amino nitrogen, ammoniacal nitorgen, electrophoresis, molecular fractionation, mineral distribution, texture characterisitics, free amino acids and free fatty acids, were checked up with the sensory test and the chesse yields at each ripening period. The results obtained by investigating the utility of Mucor rennet were summarized as follows: 1. CR chesse, MR cheese and the mixed-rennet chesse failed to show any significant difference in their yields of 15%. 2. The contents of protein, fat and ash in MR cheese gave lower value than CR cheese did and with progress of ripening lactose decreased rapidly after 14 days of ripening. The difference among the rate of addition of mucor rennet was not recognized. 3. The WSN contents of 5 fresh sample chesse were from 14.7% to 17.3% and WSN increased from 39.7% to 41.0% with progress of ripening. After 21 days of ripening MR chesse had more WSN than CR cheese did. In NCN and ammoniacal nitrogen MR cheese showed higher value. 4. As the ripening progressed, MR chesse showed more cystein, phenylalanine and proline than CR chesse did but it failed to show any increase in aspartic acid, threonine and glutamic acid etc. 5. In the content of free fatty acid MR chesse showed higher value than CR cheese did and with the progress of ripening fatty acids increased from 8.36 mEq to 26.36 mEq but did not show any significant difference in the cheese types by the coagulant ratio. 6. Ca contents in the sample chesse were 0.238-0.27%, Mg 0.019-0.022%, Na 0.910-1.047%, and K 0.175-0.200%. The important non-sedimentable Ca in casein remained from 61 % to 77% without regard the ripening periods and added-rennets and Mg remained from 59.1% to 92.5% in non-sedimentable and water-soluble conditions. 7. In the fractionation of protein by ultrafilteration, MW> $5{\times}10^4$ decresed from 95% at the beginning period of ripening to 45% and MW< $10^4$ increased from 0.2% to 38% and definite caseinolysis was shown in all samples. 8. All the cheese showed to different electrophoretic patterns for the added-amounts of mucor rennet in the 14 days of ripenig. In the 28 days or ripening, MR cheese kept some bands on the patterns compared with CR cheese. 9. In vitro digestibility increased from 81.48-94.81 % to 94.47-98.61% but failed to show any significant difference in the cheese types by the coagulant ratio. 10. In hardness, MR cheese showed lower value compared with CR cheese as the ripening progressed. 11. The results of the sensory test failed to show any difference in flora rind, feelings in mouth and hands, deep structure, flavor and bitterness between CR Camembert cheese and MR Camembert chesse.

• Tuberculosis and Respiratory Diseases
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• v.54 no.5
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• pp.551-562
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• 2003

Background : NF-${\kappa}B$ is a characteristic transcriptional factor which has been shown to regulate production of acute inflammatory mediators and to be involved in the pathogenesis of many inflammatory lung diseases. There has been some evidence that PI3K/Akt pathway could activate NF-${\kappa}B$ in human cell lines. However, the effect of PI3K/Akt pathway on the activation of NF-${\kappa}B$ varied depending on the cell lines used in the experiments. In this study we evaluated the effect of PI3K/Akt pathway on the activation of NF-${\kappa}B$ in human respiratory epithelial cell lines. Methods : BEAS-2B, A549 and NCI-H157 cell lines were used in this experiment. To evaluate the activation of Akt activation and I${\kappa}B$ degradation, cells were analysed by western blot assay using phospho-specific Akt Ab and $I{\kappa}B$ Ab. To block PI3K/Akt pathway, cells were pretreated with wortmannin or LY294002 and transfected with dominant negative Akt (DN-Akt). For IKK activity, immune complex kinase assay was performed. To evaluate the DNA binding affinity and transcriptional activity of NF-${\kappa}B$, electrophoretic mobility shift assay (EMSA) and luciferase assay were performed, respectively. Results : In BEAS-2B, A549 and NCI-H157 cell lines, Akt was activated by TNF-$\alpha$ and insulin. Activation of Akt by insulin did not induce $I{\kappa}B{\alpha}$ degradation. Blocking of PI3K/Akt pathway via wortmannin/LY294002 or DN-Akt did not inhibit TNF-$\alpha$-induced $I{\kappa}B{\alpha}$ degradation or IKK activation. Inhibition of PI3K/Akt did not affect TNF-$\alpha$-induced NF-${\kappa}B$ activation. Overexpression of DN-Akt did not block TNF-$\alpha$-induced transcriptional activation of NF-${\kappa}B$, but wortmannin enhanced TNF-$\alpha$-induced in NF-${\kappa}B$ transcriptional activity. Conclusion : PI3K/Akt was not involved in TNF-$\alpha$-induced $I{\kappa}B{\alpha}$ degradation or transcriptional activity of NF-${\kappa}B$ in human respiratory epithelial cell lines.