• Journal of Applied Biological Chemistry
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• v.54 no.2
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• pp.135-142
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• 2011

A feather-degrading bacterium Elizabethkingia meningoseptica CS2-1 was isolated from compost in a chicken farm. Cultured on a basal medium containing 2% chicken feather, the bacterium showed 729.7 ${\mu}mol/mL$ of amino acid. Optimal culture conditions for feather degradation by E. meningoseptica CS2-1 were $25^{\circ}C$, pH 7.5, and 180 rpm. The optimal pH and temperature for protease activity were 8.0 and $40^{\circ}C$, respectively. The composition of an optimal medium for amino acid production was 0.05% NH4Cl, 0.05% NaCl, 0.03% $K_2HPO_4$, 0.03% $KH_2PO_4$, 0.01% $MgCl_2{\cdot}6H_2O$, 0.1% urea, and 2% chicken feather. Characteristics of amino acids extracted from the optimal medium under the optimal culture conditions of E. meningoseptica CS2-1 were analyzed. The total amino acid content of strain CS2-1 was 1063 ${\mu}mol/mL$, which was 46% higher compared to the basal condition (729.7 ${\mu}mol/mL$). The essential amino acid content in the total amino acid was 315.9 ${\mu}mol/mL$, which was 44% higher than that of the basal condition. Major amino acids were proline (14%), aspartic acid (12%), glutamic acid (11%), serine (10%), alanine (10%), glycine (9%), and tyrosine (7%) by strain CS2-1. These results suggest that strain CS2-1 can be used as a potential microbial resource for the production of amino acid using chicken feathers.

• Korean Journal of Agricultural Science
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• v.11 no.1
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• pp.120-132
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• 1984

In order to clarify the effect of the fish meat hydrolysate on the growth of lactic acid bacteria(Str. lactis, Str. thermophibus, L. bulgaricus, L. acidophilus, and L. helveticus), the optimum conditions for hydrolyzing the fish meat were examined, and changes of the acid production, viable cell count of lactic acid bacteria and the charge of pH of the culture medium by addition of the fish meat hydrolysate were tested. The results were as follows: 1. When the hydrolysis of back muscle of mackerel was proceeded at $50^{\circ}C$ and at pH 8, for 48 hours adding 6% pancreatin of the protein content in the substrate, the best result was obtained. 2. The composition of the fish meat hydrolysate were 53.6% moisture, 32.4% protein, 1.0% fat, 10.7% carbohydrate, and 3.2% ash. 3. Above 0.1% of the fish meat hydrolysate in the culture medium, the acidity of the culture medium by Sir. lactis and Str. thermophilus were increased remarkably. The acidity of the culture medium by L. acidophilus and L. helveticus were increased in above 0.2% fish meat hydrolysate in the culture medium. but L. bulgaricus was not effected by the fish meat hydrolysate. 4. The pH of the culture medium during incubating Str. laclis and Sir. thermophilus failed obviously by adding the fish meat hydrolysate. But in the cases of L. bulgaricus, L. acidophilus, and L. helveticus, the pH were not changed clearly. 5. The viable cell count in all bacterial strains tested here were elevated by increasing the concentration of the fish meat hydrolysate.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.7
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• pp.926-934
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• 2006

The base for preparing oyster hydrolysate-added yogurt was consisted of whole milk (1,000 mL), skim milk (44.05 to 42.05 g), enzymatic oyster hydrolysates powder (OHP, 0 to 2.0 g) and pectin. The yogurt base was fermented with 7 kinds of starter cultures (3% based on yogurt volume), such as Lactobacillus acidophilus, lactobacillus bulgaricus, lactobacillus casei, Lactobacillus fermentum, Lactobacillus pentosus, Streptcoccus thermophilus and the mixed starters (L. bulgaricus and S. thermophilus) at optimal temperature. Processing condition and quality characteristics of the yogurt were evaluated by analyzing pH, titratable acidity, viscosity, viable cell count, functional properties and sensory evaluation. The results suggested that the optimal conditions for preparing the good quality yogurt revealed the mixed starters (L. bulgaricus and S. thermophilus) for starter culture, 1.0 g of 3 kDa hydrolysate for amount, and 5.5 hrs for fermentation time. The good quality yogurt showed 4.31 for pH, 1.07% for titratable acidity, 469 cps for viscosity and $4.9{\times}10^8\;CFU/mL$ for viable cell count. The hydrolysate-added yogurt was 2 times higher in ACE inhibitory and antioxidant activities than commercial yogurt, and kept good quality during storage of 15 days at $5^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.412-420
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• 1989

To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

• Journal of Animal Science and Technology
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• v.53 no.4
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• pp.349-356
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• 2011

This study was conducted to investigate the effects of brown seaweed waste (BSW) fermented with DS-01 microbe on in vitro rumen microbial fermentation. In in vitro trial, three different diets supplemented with 2%, 4%, 6% BSW fermented with DS-01 either for one month or two months was tested at 3 h, 6 h, 9 h, 12 h, and 24 h incubation. The chemical composition (CP, EE, CF, and ash) between brown seaweed waste (BSW) and fermented BSW (FBSW) were not different. The contamination of pathogenic microbes was not detected in FBSW. The pH value tended to be higher with 6% level of supplementation of FBSW for one month than other treatments. The pH at 24 h was significantly higher in FBSW than that of treatments without FBSW (p<0.05). In FBSW for two months, the pH value in 6% FBSW at 3 h in vitro fermentation tended to be higher than 2% or 4% FBSW treatments (p=0.0540), but there were no differences in other fermentation times. Although the concentration of $NH_3$-N of BSW fermented for one month was higher than control at 3 h (p<0.05), the volatile fatty acid values were significantly increased in 4 and 6% FBSW fermented for one month at 6 h incubation (p<0.05). In BSW fermented for two months, the volatile fatty acid values were significantly decreased in 6% treatment at 9 h (p<0.05). As a result of in vitro trial, it was recommended that the 2~4% supplementation level of brown seaweed waste fermented with DS-01 microbe for two months could be utilized for in vivo trial in ruminants.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.987-993
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• 2005

A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.

• Korean journal of applied entomology
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• v.16 no.4 s.33
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• pp.199-208
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• 1977

Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre.

• Korean Journal of Food Science and Technology
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• v.10 no.2
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• pp.237-244
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• 1978

Among the strains isolated form the various sources, the strain AC-12 producing a powerful pectinase was selected by the extensive screening test. The selected strain was indentified and its toxicity investigated. The conditions of the pectinase production, the characteristics of the purified enzyme and the clarification effect on the apple juice were studied. 1. The selected strain AC-12 was identified by the classification method of paper and fennel and named as Aspergillus sp. AC-12. 2. As a result of the breeding test of the white mouse, no toxicity was found from this enzyme. 3. The yield of pectinase in the medium of defatted rice bran was much better than that in the medium of wheat bran. 4. The optimum conditions for the culture of the strain in the medium of defatted rice bran were that the cultural time was 72hrs, the amount of water to be added about 80%, temperature $30{\sim}35^{\circ}C$ and pH $3.0{\sim}5.0$. 5. The yield of pectinase was slightly increased by the addition of pectin to the medium of defatted rice bran and by the addition of pectin, $NaNO_3$ and $K_2HPO_4$ to the medium of wheat bran, respectively. 6. The optimum conditions for the enzyme activity were pH $3.0{\sim}4.0$ and temperature $40{\sim}50^{\circ}C$. The enzyme was stable below $40^{\circ}C$ and pH $2.0{\sim}8.0$, respectively. But above $50^{\circ}C,$ this enzyme was abruptly inactivated. The activity was slightly increased by the addition of $MnSO_4\;and\;CuSO_4.$ 7. It was regarded that the opimum temperature for the clarification of the apple juice was $40{\sim}50^{\circ}C$, the optimum pH 3.0 and the optimun concentration of the enzyme 0.1%, and the apple juice was almost clarified by the reaction at $45^{\circ}C$ for 60 minutes.

• Applied Biological Chemistry
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• v.11
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• pp.83-88
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• 1969

94 Cellulase producing strains were isoated from soils, composts, rotten woods and straws, and gastric contents and feces of herbivorous animals in various places. Among them, the strain MC-9, MC-10, MC-53 and MC-61 were found to be highly active in the degradation of carboxy methyl cellulose. Their cultural conditions adequate for the cellulase formation and effects of inorganic salts and various organic substances added to the wheat bran media were investigated. The results obtained are as follows; 1. Optimum conditions for the cellulase formation were MC-9: pH 5.5, temp. $35^{\circ}C$, incubation time 5 days, MC-10: pH 5.5-6.0, temp. $30^{\circ}C$, incubation time 5 days, MC-53: pH 3.5, temp. $30^{\circ}C$, incubation time 5 days, MC-61: pH 3.5-4.0, temp. 30-$35^{\circ}C$, incubation time 5 days. 2. Their cellulase activity in their optimum conditions were MC-9: CMC-LP(liquefying power). 87.7%, CMC-SP(saccharifying power) 3.20 glucose mg./gm. of the cultures/min., MC-10: CMC-LP 82.9%, CMC-SP 2.48 glucose mg./gm. of the cultures/min., MC-53: CMC-LP 72.4%, CMC-SP 1.76 glucose mg./gm. of the cultures/min., MC-61: CMC-LP 87.1%, CMC-SP 2.08 glucose mg./gm. of the cultures/min. 3. Additions of inorganic salts to the wheat bran media were not significant for the cellulase formation, but additions of soybean film and orange-peel pomace promoted the CMC-liquefying power 3 to 5 percent in wheat bran cultures of the strains.

• Journal of Life Science
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• v.18 no.4
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• pp.530-536
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• 2008

To investigate the molecular mechanism of the airway inflammation by Pseudomonas fluorescens, effects on the inflammatory mediators such as interleukin-8 (IL-8), cyclooxygenase-2 (COX-2), macophage inhibitory cytokine 1 (MIC-1) were assessed in the human alveolar epithelial cells. Exposure to P. fluorescens and its recombinant bacteria suppressed cellular viability in the A549 epithelial cells and pro-inflammatory cytokine interleukin-8 production. However, pro-inflammatory prostaglandin-producing COX-2 protein was not altered by P. fluorescens though its mRNA was slightly elevated. As the inhibitory cytokine for the pro-inflammatory mediators, MIC-1 expression was monitored in A549 cells. MIC-1 gene induction was not significantly enhanced but the protein processing was changed by exposure to P. fluorescens. Pro-protein form of MIC-1 (${\sim}40\;kD$) was cleaved into active form mature MIC-1 (${\sim}15\;kD$) and propeptide (${\sim}28\;kD$) by the bacteria exposure. MIC-1 activation can contribute to the suppression of cellular viability by P. fluorescens and can retard IL-8-induced monocyte recruitment. However, sustained activation of MIC-1 can mediate the tissue injury by P. fluorescens exposure.