• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.506-508
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• 1986

Metabolic activity of inorganic nitrogenous compounds affects not only microbial growth but also metabolite production in fermentation technology. We have worked on the enzymes participating in ammonia assimulation of some fermentation bacteria. This paper summarizes the results on glutamine synthetase and its application in practical field. Glutamine synthetase (L-glutamate:ammonia ligase, EC. 6.3.1.2) catalyzes the formation of glutamine from glutamate and ammonia at the expense of cleavage of ATP and inorganic phosphate. The enzyme plays a dual role in nitrogen metabolism in bacteria; it is a key enzyme not only in the biosynthesis of various compounds through glutamine but also in the regulation of synthesis of some enzymes involved in the metabolism of nitrogenous compounds. The detailed works with the Eschericia coli and other enterobacterial enzymes revealed that glutamine synthetase is controlled by the following complex of mechanisms: (a) feedback inhibition by end products, (b) repression and derepression of enzyme synthesis, (c) modulation of enzyme activity in response to divalent cation and (d) covalent modification of enzyme protein by adenylylation and its cascade control. Comparative studies have also been made on the enzymes from other organisms.

• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.75-80
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• 1995

Dextranase produced by Flavobacterium multivorum HL-1 isolated from soil were characterized. Optimum growth condition for the production of the enzyme by the strain appeared to be at the 1.0% concentration of dextran. When NH$_{4}$N0$_{3}$ and beef extract was added to the culture media, the enzyme activity increased upto 570 and 680 units per ml respectively, and other nitrogen sources did not increase the activity. Urea, casamino acid, (NH$_{4}$)$_{2}$SO$_{4}$ and KNO$_{3}$ inhibited the activity. In the presence of Mg^{2+} in culture media, the enzyme activity increased 124%, but its activity was inhibited in the presences of Ca$^{2+}$, Co2$^{2+}$, Hg^{2+} and Zn$^{2+}$ . The optimum temperature for the enzyme activity was 45-55$\CIRC$C . In the ranges of pH 4 to 10, the activity of the enzyme appeared approximately similar.

• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.652-656
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• 1996

The 80 strains which produce carrageenan degrading enzyme were isolated from soils, mud, seaweed, marine moluscus and echonodermata samples. Among them, one isolated strain, which showed the highest activity to produce carrageenan degrading enzyme, was used for this study. The isolated strain was identified as Pseudomonas alcaligenes through its morphological, biochemical, and physiological characteristics. The best conditions for enzyme production were 0.7% nutrient broth and 0.2% carrageenan as nitrogen and carbon source, respectively. The optimal pH, NaCl, temperature and culture time for carrageenan degrading enzyme were 7.0, 1.5%, 30* and 96hrs, respectively.

• Journal of Ginseng Research
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• v.3 no.2
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• pp.134-143
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• 1979

It order to extract the soluble organic substances of Korean ginseng effectively, the ginseng extract have been made by using amylase. The investigation on the optimum condition of enzyme reaction was carried out, and the amounts of gained extract and its saponin pattern were compared among the ethanol extract, water extract and enzyme extract. The results obtained are summerized as follows. 1. The gaining ratio or ginseng extract was the highest value when the raw ginseng and dried ginseng were extracted in the concentration of 7.5% and 5% with 0.3%∼0.6% enzyme for 25 hour. 2. The amounts of ethanol extract, water extract and enzyme extract were 9.14%, 17.23% and 23.73% in case of raw ginseng and 64.09%, 72.52% and 74.36% in case of dried ginseng, respectively. The amount of enzyme extract was increased as much as 6∼14% in case of raw ginseng, and 2∼10% in case of dried ginseng compared with that of ethanol and water extract. 3. The absolute content of saponin was nearly constant in spite of the different extraction method and all of the ginseng saponin pattern of thin-layer chromatograms were almost same.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.737-748
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• 2010

Glutaminase production in Zygosaccharomyces rouxii by solid-state fermentation (SSF) is detailed. Substrates screening showed best results with oatmeal (OM) and wheatbran (WB). Furthermore, a 1:1 combination of OM:WB gave 0.614 units/gds with artificial sea water as a moistening agent. Evaluation of additional carbon, nitrogen, amino acids, and minerals supplementation was done. A central composite design was employed to investigate the effects of four variables (viz., moisture content, glucose, corn steep liquor, and glutamine) on production. A 4-fold increase in enzyme production was obtained. Studies were undertaken to analyze the time-course model, the microbial growth, and nutrient utilization during SSF. A logistic equation ($R^2$=0.8973), describing the growth model of Z. rouxii, was obtained with maximum values of ${\mu}_m$ and $X_m$ at $0.326h^{-1}$ and 7.35% of dry matter weight loss, respectively. A goodfit model to describe utilization of total carbohydrate ($R^2$=0.9906) and nitrogen concentration ($R^2$=0.9869) with time was obtained. The model was used successfully to predict enzyme production ($R^2$=0.7950).

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.5 no.1
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• pp.137-150
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• 1999

Macrophage play a major role in host defence against infection and tumor development and this activity is regulated through the production of several mediators. In particular, the production of NO by macrophages mediates killing or growth inhibition of tumor cells, bacteria, fungi and parasites. However, over-expression of iNOS by various stimuli, resulting in over-production of NO, contributes to the pathogenesis of septic shock and some inflammator and auto-immune disease. Therefore, it would be valuable to develop potent and selective inhibitors of for potential therapeutic use. Thus the agent that supprees the expression of iNOS mRNA or enzyme protein will be usefull for the prevention of various diseases. We are intersted in identifying selective inhibitiors of iNOS which might be useful intreating inflammatory human diseases. In summary, we have demenstrated that scopoletin, isolated from Seman Armenicae and Radix Trichosanthis the production of NO induced by $IFN-\gammer$ plus LPS in RAW 264.7 macrophages, The mechanism for the inhibition of NO production was due to suppression of the expression of iNOS mRNA or enzyme protein.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.209-218
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• 2015

Fructooligosaccharides have been mainly produced by microbial fructosyltransferases (FTase) enzymes. The present work focuses on the optimization of medium composition and cultivation parameters affecting FTase produced by Penicillium aurantiogriseum AUMC 5605 in shake flask cultivation. FTase production was optimized in two steps using DeMeo's fractional factorial design. A 1.46-fold increase in FTase production (105.4 U/mL) was achieved using the optimized culture medium consisting of (g/L): sucrose, 600; yeast extract, 10; $K_2HPO_4$, 5; $MgSO_4{\cdot}7H_2O$, 0.5; $(NH_4)_2SO_4$, 1.0 and KCl, 0.5. The obtained results showed that the maximum FTase enzyme activity was produced at initial cultivation pH values ranging from 6.0-6.5, at agitation speed of 200 rpm and using vegetative fungal cells as inoculum. Moreover, results showed that optimization of medium composition and some cultivation parameters resulted in an increase of about 93.7% in the enzyme activity than the nonoptimized cultivation conditions after 96 h of cultivation. Additionally, maximum production and specific production rates recorded 2340 U/L/h and 102 U/L/h/g cells, respectively.

• Korean Journal of Food Science and Technology
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• v.50 no.5
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• pp.528-534
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• 2018

The current study investigated the effects of enzyme treatment on black rice (Oryza sativa L.) aleurone layer extracts. Different enzymes (lipase, lecitase ultra and lipopan 50BG) were used to test anti-oxidant and anti-inflammatory activities in vitro. The antioxidant activities of enzyme treated or non-enzyme treated extracts of black rice bran were evaluated via 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity. Lipase treated extracts of black rice bran showed higher antioxidant activity compared to that of non-enzyme treated extracts. Anti-inflammatory activities of enzyme treated black rice bran extracts on nitrite production and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) secretion, were tested using a nitric oxide (NO) colorimetric assay kit and an enzyme-linked immunosorbent assay (ELISA) kit. The ethanolic extract of enzyme treated black rice bran decreased the levels of nitric oxide production and pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$, in a lipopolysaccharidestimulated RAW 264.7 cell culture. These findings indicate that enhanced anti-oxidant and anti-inflammation activities of the ethanolic extracts of enzyme treated black rice (Oryza sativa L.) aleurone layers, may be attributed to molecular conversion of ingredients in enzyme catalyzed reactions.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1742-1750
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• 2015

Human insulin is composed of 21 amino acids of an A-chain and 30 amino acids of a B-chain. This is the protein hormone that has the role of blood sugar control. When the recombinant human proinsulin is expressed in Escherichia coli, a serious problem is the formation of an inclusion body. Therefore, the inclusion body must be denatured and refolded under chaotropic agents and suitable reductants. In this study, H27R-proinsulin was refolded from the denatured form with β-mercaptoethanol and urea. The refolding reaction was completed after 15 h at $15^{\circ}C$, whereas the reaction at $25^{\circ}C$ was faster than that at $15^{\circ}C$. The refolding yield at $15^{\circ}C$ was 17% higher than that at $25^{\circ}C$. The refolding reaction could be carried out at a high protein concentration (2 g/l) using direct refolding without sulfonation. The most economical and optimal refolding condition for human preproinsulin was 1.5 g/l protein, 10 mM glycine buffer containing 0.6 M urea, pH 10.6, and 0.3 mM β-mercaptoethanol at $15^{\circ}C$ for 16 h. The maximum refolding yield was 74.8% at $15^{\circ}C$ with 1.5 g/l protein. Moreover, the refolded preproinsulin could be converted into normal mature insulin with two enzymes. The average amount of human insulin was 138.2 g from 200 L of fermentation broth after enzyme reaction with H27R-proinsulin. The direct refolding process for H27R-proinsulin was successfully set up without sulfonation. The step yields for refolding and enzyme reaction were comparatively high. Therefore, our refolding process for production of recombinant insulin may be beneficial to the large-scale production of other biologically active proteins.

• Journal of the Korean Wood Science and Technology
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• v.23 no.4
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• pp.108-116
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• 1995

This experiment was conducted to screen a superior wood-rotting fungi for lignin degradation and ligninolytic enzyme production by evaluation of red colored zone width on potato-dextrose agar medium and oak woodmeal medium complimented guaiacol. Relationship between the red colored zone width on GU-WA medium and klason lignin loss on woodmeal medium showed the positive correlation. Thus, the potential ligninolytic activity of wood rotting fungi which are not elucidated yet may be estimated to some extent by the evaluation of the red colored zone width on GU-WA medium. Of the isolates screened from fruit bodies and decayed woods. LKY-12, LKY-7 and C. versicolor-13 isolates having preferential lignin degradation and laccase activity were selected. These isolates exhibited characteristics of superior wood-rotting fungi as Klason lignin loss ranged from 30% to 35% and ligninolytic enzyme activity of these isolates on glucose-peptone broth was higher than that of other isolates. And then, these isolates were considered to be able to use in biological pulping and bleaching and ligninolytic enzyme production.