• Applied Biological Chemistry
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• v.27
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• pp.29-46
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• 1984

To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.

• Korean Journal of Food Science and Technology
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• v.8 no.1
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• pp.19-32
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• 1976

The laboratory Mejus as well as home-made Meju and improved Meju received from Korea were ripened in the brine for up to 8 months and the changes is the chemical composition during the process were determined and the differences between the types of Meju were compared. On the basis of the amino acid pattern, the changes in the protein quality of soybean during the process was evaluated. No significant changes in the general chemical composition of Meju were noticed during the ripening for 8 months. However, the nitrogen solubility of Meju increased for $13{\sim}29%$ to $66{\sim}78%$ during 8 month ripening of the Meju-brine mixture. The concentration of free amino-N to the total-N increased from $4{\sim}7%$ in Meju to $29{\sim}35%$ in the 8month ripened mixture. The concentration of amino-N to the total-N increased from $1{\sim}4%$ in Meju to $5{\sim}14%$ in the 8month ripened mixture and the changes varied with the type of Meju used. Remarkable changes in the amino acid pattern of soybean were occured during the ripening process. The concentration of methionine decreased to the half of original Meju during the first month of ripening. Arginine and histidine were destroyed rapidly by the ripening longer than 1 month. A considerable amount of ornithine was synthesized during the ripening. The amino acid pattern of Meju did change drastically during the ripening longer than 3 months and the changes varied with the type of Meju. The retention of the nutrients in soybean during 8 month ripening of the laboratory 3 month Meju in the brine was 49% for carbohydrates, 107% for crude fat, 93% for crude protein and 74% for the total amino acid. Histidine, arginine and methionine and 74% for the total amino acid. Histidine, arginine and methionine were the most damaged during the process, retaining only 25%, 27% and 49% of the contents in raw soybean, respectively, whereas lysine retained 79%. By the separation of the 8 month ripened mixture, approximately 60% of crude protein, all of crude fat and 80% of carbohydrates in the mixture were retained in soypaste. Soypaste contained higher concentrations of amino acids per 16gN compared to soysauce, except for lysine. The most limiting amino acid of the protein was the S-containing amino acids in all cases studied, whereas the second limiting amino acid varied from valine in soybean to threonine in most of Mejus and the brine mixtures, lysine in most of soypastes and tryptophan in some of soysauces. According to the protein quality evaluation made by the reference of the FAO provisional pattern of amino acid, the chemical score of raw soybean was 82, which was reduced to 77 by cooking and further reduced to $71{\sim}74$ by Meju fermentation. At the eighth month of ripening the chemical score of the Meju-brine mixtures were reduced to $51{\sim}66. After the separation, the chemical score of soypaste ranged from 60 to 71, whereas that of soysauce varied from 45 to 57. Generally, the products made from improved Meju recorded the highest score, whereas those made from homemade Meju showed the poorest protein quality. The essential amino acid index(EAAI) of the samples was similar to the chemical score, but it appeared to fit the overall changes in the amino acid pattern during the process better than the chemical score.

• Applied Biological Chemistry
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• v.14 no.1
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• pp.59-97
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• 1971

As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

• Tuberculosis and Respiratory Diseases
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• v.42 no.5
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• pp.744-751
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• 1995

Background: Asthma is an inflammatory disease because there are many inflammatory changes in the asthmatic airways. Axon reflex mechanisms may be involved in the pathogenesis of asthma. Sensory neuropeptides are involved in this inflammation, which is defined as neurogenic inflammation. Substance p, neurokinin A, and neurokinin B may be main neuropeptides of neurogenic inflammation in airways. These tachykinins act on neurokinin receptors. Three types of neurokinin receptors, such as $NK_1$, $NK_2$, and $NK_3$, are currently recognized, at which substance p, neurokinin A, and neurokinin B may be the most relevant natural agonist of neurogenic inflammation in airways. The receptor subtypes present in several tissues have been characterized on the basis of differential sensitivity to substance p, neurokinin A, and neurokinin B. Plasma extravasation and vasodilation are induced by substance p more potently than by neurokinin A, indicating NK1 receptors on endothelial cells mediate the response. But airway contraction is induced by neurokinin A more potently than by substance P, indicating the $NK_2$ receptors in airway smooth muscles. These receptors are used to evaluate the pathogenesis of brochial asthma. FK224 was identified from the fermentation products of Streptomyces violaceoniger. FK224 is a dual antagonist of both $NK_1$ and $NK_2$ receptors. Purpose: For a study of pathogenesis of bronchial asthma, the effect of FK224 on plasma extravasation induced by vagal NANC electrical stimulation was evaluated in rat airway. Method: Male Sprague-Dawley rats weighing 180~450gm were anesthetized by i.p. injection of urethane. Plasma extravasation was induced by electrical stimulation of cervical vagus NANC nerves with 5Hz, 1mA, and 5V for 2 minutes(NANC2 group) and for sham operation without nerve stimulation(control group). To evaluate the effect of FK224 on plasma extravasation in neurogenic inflammation, FK224(1mg/kg, Fujisawa Pharmaceutical Co., dissolved in dimethylsulphoxide; DMSO, Sigma Co.) was injected 1 min before nerve stimulation(FK224 group). To assess plasma exudation, Evans blue dye(20mg/kg, dissolved in saline) was used as a plasma marker and was injected before nerve stimulation. After removal of intravascular dye, the evans blue dye in the tissue was extracted in formamide($37^{\circ}C$, 24h) and quantified spectrophotometrically by measuring dye absorbance at 629nm wavelength. Tissue dye content was expressed as ng of dye per mg of wet weight tissue. The amount of plasma extravasation was measured on the part of airways in each groups. Results: 1) Vagus nerve(NANC) stimulation significantly increased plasma leakage in trachea, main bronchus, and peripheral bronchus compared with control group, $14.1{\pm}1.6$ to $49.7{\pm}2.5$, $17.5{\pm}2.0$ to $38.7{\pm}2.8$, and $12.7{\pm}2.2$ to $19.1{\pm}1.6ng$ of dye per mg of tissue(mean ${\pm}$ SE), respectively(p<0.05). But there was not significantly changed in lung parenchyma(p>0.05) 2) FK224 had significant inhibitory effect upon vagal nerve stimulation-induced airway plasma leakage in any airway tissues of rat,such as trachea, main bronchus, and peripheral bronchus compared with vagus nerve stimulation group, 49%, 58%, and 70%, respectively(p<0.05). Inhibitory effect of FK224 on airway plasma leakage in neurogenic inflammation was revealed the more significant in peripheral bronchus, but no significant in lung parenchyma. Conclusion: These results suggest that FK224 is a selective NK receptor antagonist which effectively inhibits airway plasma leakage induced by the endogenous neurotransmitters relased by neurogenic inflammation in rat airway. Tachykinin receptor antagonists may be useful in the treatment of brochial asthma.

• Korean journal of food and cookery science
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• v.22 no.5 s.95
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• pp.642-649
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• 2006

The browning of soy sauce is caused by the reaction of amino-carbonyl between amino-compounds and reducing sugar. Only a few studies have investigated the formation of melanoidins in UHYUKJANG. The objectives of this study were to analyze the brown pigment of UHYUKJANG and to investigate the characteristics of UHYUKJANG in comparison with soy sauce and model melanoidins. The samples were ripened for 0, 60, 120, 180, 240, 300 and 360 days at 4$^{\circ}C$ and 20$^{\circ}C$. The pH, absorbance at 420 nm absorbance ratio of 400 to 500 nm and UV-VIS spectra as an index of color intensity were measured. Additionally, L, a and b values of the samples and the amount of 3-Deoxyglucosone(3DG) in the samples were measured. The pH of both soy sauce (from 6.26 to 5.52) and UHYUKJANG (from 6.13 to 5.11) rapidly decreased during the first 60 days of aging and was also affected by storage temperature. The absorbance of samples at 420 nm increased during the aging process, reaching its maximum after 180 days, regardless of sample and temperature. On the other hand, the intensity of brown color in the samples increased with increasing aging period according to the results of absorbance ratio (soy sauce: 1.37 to 5.29, UHYUKJANG: 1.37 to 5.02). The L value of soy sauce increased during the aging process and was maximized after 240 days at 4$^{\circ}C$ and 180 days at 20$^{\circ}C$, but decreased thereafter. There was no significant difference in L value of UHYUKJANG, regardless of aging period and temperature. On the other hand, the b value did not reveal any significant change during aging, but the a value increased until 120 days of aging in the other samples except for UHYUKJANG at 20$^{\circ}C$. The average amount of 3DG separated from soy sauce was 5.65 mg%, and from UHYUKJANG was 3.74 mg%. These results indicated that the browning of UHYUKJANG was also caused by melanoidins produced by the reaction of amino-carbonyl during the fermentation process.

• The Korean Journal of Food And Nutrition
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• v.24 no.3
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• pp.367-375
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• 2011

For this study, Korean-type Koumiss was made by the fermentation of mixed cultures, in which yeast, Kuyveromyces, and microflora, Streptococcus thermophiles and Lactobacillus bulgaricus, were inoculated into 10% skimmed milk with added whey powder(control: A, 2%: B, 4%: C, 6%: D, and 8%: E). Fat, protein, lactose, titratable acidity, pH, the number of lactic acid bacteria, the number of yeast, alcohol content, volatile fatty acids, volatile free amino acids and minerals were measured in the products. The results were as follows: As the dosage of whey powder increased, fat increased from 0.74% in the control to 2.30% in sample E, protein increased from 2.95% in the control to 4.39% in sample E and lactose increased from 3.10% in the control to 7.43% in sample E. Titratable acidity and pH increased gradually. The number of lactic acid bacteria increased from $10^9\;cfu/m{\ell}$ in the control to $3.8{\times}10^9\;cfu/m{\ell}$ in sample E, and the number of yeast increased from $6.1{\times}10^7\;cfu/m{\ell}$ in the control to $1.65{\times}10^8\;cfu/m{\ell}$ in sample E, according to the increase of whey powder content. For alcohol content, the average values were 0.863%, 0.967%, 0.890%, 1.290%, and 1.313% for the control and samples B, C, D, and E, respectively. As the dosage of whey powder increased, alcohol content showed a tendency to gradually increase. The average alcohol content of E was 1.313 and this was higher than the alcohol content of Kazahstana-type Koumiss with 1.08%. Sixteen types of free amino acids were detected. Glycine was the lowest in the control at $0.38mg/m{\ell}$ and sample E contained $0.64mg/m{\ell}$. Histidine was also low in the control at $0.42mg/m{\ell}$ and sample E contained $0.65mg/m{\ell}$. On the other hand, glutamic acid was highest at $4.13mg/m{\ell}$ in the control whereas sample E had $6.96mg/m{\ell}$. Proline was also high in the control at $1.71mg/m{\ell}$ in control, but E contained $2.80mg/m{\ell}$. Aspartic acid and leucine were greater in sample E than in the control. For volatile free fatty acids, content generally had a tendency to increase in the control, and samples B, C, D, and E. Content of acetic acid gradually increased from $12,661{\mu}g/100m{\ell}$ in the control to $37,140{\mu}g/m{\ell}$ in sample E. Butyric acid was not detected in the control and was measured as $1,950{\mu}g/100m{\ell}$ in sample E. Caproic acid content was $177{\mu}g/100m{\ell}$ in the control and $812{\mu}g/100m{\ell}$ in sample E, and it increased according to the increase of whey powder content. Valeric acid was measured in a small amount in the control as $22{\mu}g/100m{\ell}$, but it was not detected in any other case. Mineral contents of Ca, P, and Mg increased from 1,042.38 ppm, 863.61 ppm, and 101.28 ppm in the control to 1,535.12 ppm, 1,336.71 ppm, and 162.44 ppm in sample E, respectively. Na content was increased from 447.19 ppm in the control to 1,001.57 ppm in sample E. The content of K was increased from 1,266.39 ppm in the control to 2,613.93 ppm in E. Mineral content also increased with whey powder content. In sensory evaluations, the scores increased as whey powder content increased. Flavor was lowest in the control with 6.3 points and highest in E with 8.2 points. Body and texture were highest at 4.2 points in the control, which did not have added whey powder. In the case of appearance, there were no great differences among the samples.