• Title/Summary/Keyword: total substrate inhibition concentration

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Theoretical Consideration of the Modified Haldane Model of the Substrate Inhibition in the Microbial Growth Processes (미생물 성장 공정에서의 기질 저해에 관한 modified Haldane 모델의 이론적 고찰)

  • Hwang, Young-Bo
    • Applied Chemistry for Engineering
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    • v.19 no.3
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    • pp.277-286
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    • 2008
  • This paper deals with the theoretical derivation of the modified Haldane model of the substrate inhibition in the microbial growth processes. Based on the biological concepts of substrate-receptor complex working mechanisms, a new microbial kinetics of N-fold multiplex substrate inhibition and its generalization has been considered theoretically, which is natural expansion of the simple substrate inhibition mechanism in the enzyme reaction. As a result, the modified Haldane model of the substrate inhibition turns out to be a well-designed four-parameter kinetic model with a biological constant of the total substrate inhibition concentration.

Production of L-Tryptophan by Enzymatic Processes (효소공정에 의한 트립토판 생산)

  • 이인영;안경섭;김의환;이선복
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.73-78
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    • 1992
  • - Enzymatic synthesis of L-tryptophan(Trp) using E. coli tryptophanase has been investigated. In order to reduce the substrate inhibition by indole and to increase the product yield of L-tryptophan three different approaches have been made in this work. First, indole was intermittently fed to the reaction mixture in order to control the indole concentration at lower level. When 15 mM of indole was used as a total amount of substrate, conversion yield of 80% has been obtained with intermittent feeding while only 20% of indole was converted into L-tryptophan by conventional batch operation, The second method employed in this work was the use of cyclohexane-phosphate buffer organic two-phase system. In this system, indole was mainly partitioned into the organic-solvent phase and therefore substrate inhibition was expected to be reduced. L-Tryptophan production in organic two-phase system was, however, unexpectedly lower than that obtained in aqueous buffer solution. As a third method cyclodextrins have been added to the aqueous reaction mixture. It was found that the addition of $\beta$-cyclodextrin enhanced the tryptophan synthesis noticeably while $\alpha$-cycfodextrin showed little effect on tryptophan production.

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Enzymatic Hydrolysis of Pretreated Chitin by Aspergillus carneus Chitinase

  • Mohamed, Abdel-Naby;Kwon, Dae-Young
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.197-203
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    • 1992
  • Studies of the pretreatment of chitin and its subsequent hydrolysis by Aspergillus carneus chitinase are reported. Ball milling was found to be the most effective way among the pretreatment methods tested. Data are presented describing the effect of enzyme and substrate concentrations on the rate and extent of the hydrolysis process. It was found that the successive addition of enzyme improved the saccharification yield. Significant product inhibition of the chitinase was observed when N-acetylglucosamine concentration was 3.6% or higher. Adsorption of enzymes to the substrate occurred during a 24 hr hydrolysis period. An initial rapid and extensive adsorption occurred, followed by gradual desorption which increased during the time of reaction. Intermediate removal of the hydrolyzate and continuation of the hydrolysis by adsorbed enzyme on the residual chitin was also investigated. A total of 75.4 g/l reducing sugars, corresponding to 69.2% saccharificaton yield (as N-acetylglucosamine) was obtained. In addition an increase in the amount of recoverable enzymes was observed under these conditions. Evidence presented here suggests that the technique, whereby the free enzymes in the recovered hydrolyzate are re-adsorbed onto the new substrate, may provide a means of recirculating the dissolved enzymes.

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Effect of seeding ratio on acidogenic biokinetics in high ammonia concentration

  • Yang, Keun-Young;Shin, Seung-Gu;Hwang, Seok-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.65-66
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    • 2005
  • Anaerobic digestion is one of the well-known methods for biological treatment handling of concentrated organic matter such as swine $wastewater.^{1)} The anaerobic digestion can reduce organic loading but also hydrolyze non-biodegradable organic $matter.^{2)}$ The feces from the scrapper-type barn are usually collected to make compost and the urine is discarded with swine-slurry wastewater by ocean-dumping or treated by biological methods. The lagoon, aerobic digestion, anaerobic digestion, SBR, $A^{2}/O$, and UCT have been applied for treating swine $wastewater.^{3)} In this study, as a result of the analysis of swine wastewater, the total and soluble chemical oxygen demand was 130g/L and 60g/L, respectively. And the volatile fatty acid as chemical oxygen demand equivalent was 45g/L, which was 75% of soluble chemical oxygen demand. Before everything else, ammonia nitrogen concentration was 6.5 g/L. From biochemical acidogenic potential test, it was concluded that the enhanced acidification process to manage swine waste should be operated in the ammonia nitrogen concentration of less than 1.2 g/L. In the result of seeding ratio experiments with artificial $wastewater^{4)}, the lag period of acidogens was taken the long time because of the inhibition by the $ammonia^{5)}$, however no difference of period by the seeding ratio was not shown. The Haldane-based biokinetics were also evaluated using a method of fourth order Runge-Kutta $approximation.^{6,7)}$ The nonlinear least squares (NLLS) method with a 95% confidence interval was also used. The ranges of maximum microbial growth rate, ${/mu_{max}}$, and half saturation coefficient, $K_{s}$, for acidogenesis of various seeding ratio with artificial wastewater were 6.1 ~ 12.6 $d^{-1}$ and 45,000 ~ 53,500 mg glucose/L, respectively. Also, the methanogenic microbial yield coefficient, Y, and microbial decay rate coefficient, $k_{d}$, and inhibition substrate concentration, $K_{si}$, for the reactors were determined to be 0.32 ~ 0.465 ${/mu}g$/mg glucose; 0.42 ~ 1.01 $d^{-1}$ and 51,500 ~ 55,600 mg glucose/L, respectively.

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Effect of Ginseng Saponin on Bacterial α-Amylase Activity (인삼(人蔘) Saponin이 세균(細菌) α-Amylase 활성(活性)에 미치는 영향(影響))

  • Do, Jae Ho;Kim, Sang Dal;Joo, Hyun Kyu
    • Microbiology and Biotechnology Letters
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    • v.13 no.1
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    • pp.7-11
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    • 1985
  • In order to investigate the biological activity of ginseng saponins, the effects of ginseng saponins on the reaction catalyzed by bacterial a-amylase were studied and the results obtained were summerized as follows. Bacterial ${\alpha}$-amylase activity was increased by the addition of protopanaxadiol (diol), protopanaxatriol (triol) and total saponin. Preincubation of ${\alpha}$-amylase with diol saponin at $40^{\circ}C$ for 3 min increased ${\alpha}$-amylase activity to the degree of 120%. In the protective effect on the heat denaturation of the enzyme, triol saponin protected the heat denaturation for 5 min at $60^{\circ}C$, but diol saponin accelerated the heat denaturation. The hydrolyzates of diol and triol saponin increased the enzyme activity more than the intact diol and triol saponin. In the catalysis system of bacterial ${\alpha}$-amylase, the addition of diol and triol saponin reduced the substrate inhibition in the presence of high concentration of the substrate.

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Fed-Batch Culture of Brevibacterium CHI for the Production of Nitrile Hydratase (Brevibacterium CH1의 유가 배양에 의한 Nitrile Hydratase의 생산)

  • 황준식;황영보;이처영;장호남
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.614-618
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    • 1992
  • The batch and fed-batch cultivations of Brevibacteriurn CHI were carried out for the production of nitrile hydratase. In batch culture, with pH control the cell mass and the specific activity increased more 20% and 30%. respectively. The maximum growth rate was obtained at a glucose concentration of $20g/{\ell}$ because of substrate inhibition. The fed-batch culture of Brevibacteriurn CHI with constant substrate feeding gave a cell density of up to $68g/{\ell}$ and nitrile hydratase activity was maintained at above 6.1units/mg. The cell growth yield on carbon .source was ca. 0,68 g/g glucose consumed. The total nitrile hydratase activity in this fed-batch mode increased up to 414.8 units/m${\ell}$, which amounted to 4.4 times that of the batch culture.

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Hydrolysis of Fish Protein Concentration in an Ultrafiltration Membrane Reactor (한외여과막 반응기를 이용한 FPC의 가수분해)

  • 최정호;변희국;김세권
    • Membrane Journal
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    • v.10 no.2
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    • pp.83-91
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    • 2000
  • In order to improve functional properties, enzymatic hydrolysis of FPC (fish protein concentration) was achieved in ultrafiltration membrane reactor (MWCO 5,000). First, insoluble FPC was hydrolyzed by pepsin in batch reactor to decrease the fouling in ultrafiltration membrane reactor, and second hydrolysis was achieved by pronase E in ultrafiltration membrane reactor The optimum operating conditions in batch reactor using pepsin were at temperature 45$^{\circ}C$, pH 2.0 and the ratio of substrate to pepsin, 150 (w/w) After operating for 5hrs under optimum conditions, 89% of total amount of initial FPC was hydrolyzed. The rate constants, $K_{m}$ and V$_{max}$, were 1.25% and 0.89 mg/$m\ell$/min, respectively, and substrate inhibition was occured above 1.5%. The ultrafiltration membrane reactor was operated with recycling rate of 474 $m\ell$/min and transmembrane pressure of 15 psi. The permeate flux was increased by temperature, transmembrane pressure, but the permeate flux was fixed by pH. The optimum ratio of substrate to pronase E was 200(w/w) and the productivity of ultrafiltration membarane reactor was 702 mg/mg -enzyme, that of batch reactor was 51mg/mg-enzyme. Molecular weight distributions tot first and second hydrolysates were from 2,500 Da to 20,000 Da and from 700 Da to 10,000 Da, respectivelyly.

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Anti-bacterial Effects of Aqueous Extract Purified from the Immature Cone of Red Pine (Pinus densiflora) (미성숙 솔방울 열수추출물의 항균성)

  • Jeong, Kyung Hui;Hwang, In Sik;Kim, Ji Eun;Lee, Young Ju;Kwak, Moon Hwa;Lee, Young Hee;Lee, Jae Ho;Hwang, Dae Youn;Jung, Young Jin
    • Textile Coloration and Finishing
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    • v.26 no.1
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    • pp.45-52
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    • 2014
  • Red pine (Pinus densiflora) is widely used traditional medicine, pharmacological and nutritional values from which the phytochemical compounds are derived. The present study was aimed to examine the antibacterial effects in the absence and presence of a immature red pine cone extract against 13 microorganisms. The components in the aqueous extract from immature red pine cone were identified by GC-MS. About 1.4% of total polyphenolic compound was measured in aqueous extract collected from immature red pine cone. Also, the high concentration of ${\beta}$-phellenandrene, ${\alpha}$-pinene, limonene, bornyl acetate and aldehyde was detected in total ion chromatograms. Of total 13 microorganisms, 4 microorganisms including Pseudomonas aeruginosa, Vibrio cholera, Listeria monocytogenes, Klebsiella pneumonia were effectively killed by aqueous extract of immature red pine cone. The highest anti-bacterial effect was detected in P. aeruginosa, followed by V. cholera, L. monocytogenes and K. pneumonia. In case of P. aeruginosa, the largest diameter of inhibition zone was maintained to 1/2 solution treated cells and slightly decreased at 1/4 and 1/8 solution treated cells. Also, in test used V. cholera and L. monocytogenes, the inhibition zone was strongly formed in only 1 and 1/2 solution treated cells, while K. pneumonia showed the very small diameter of inhibition zone in all concentrations. Therefore, these results suggested that the aqueous extracts of immature red pine cone should be considered as a new and potentially important anti-bacterial substrate to effectively prevent the microbial infection and penetration.

Studies on the Naringinase of Mold-[part 1] Screening test of Molds on the Production of Naringinase and some properties of Crude Enzyme of Selected strain- (사상균 Naringin 분해 효소에 관한 연구-[제1보] 우량 균주의 분리 선별과 선별균의 조효소 성질에 관하여-)

  • Ki, Woo-Kyung;Sung, Nack-Kie
    • Applied Biological Chemistry
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    • v.13 no.3
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    • pp.237-242
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    • 1970
  • Fifty strains of mold which isolated from the various sources were screened for the production of Naringinase which hydrolyse naringin, the 7-rhamnoe-glucoside of 4'.5.7. - trihydroxyflavanonin, the main bitter principle of citrus fruits and grape fruits. Of the 4 strains yielded naringinase with significant activity, S-1 strain was selected on the criterion of industrial application, and some properties of crude naringinase of this S-1 was investigated. The results obtained were as follows. 1. Naringenase obtained from S-1 strain has optimum pH range from 3.0 to 5.0 for its activity. 2. Production of naringinase was increased on the addition of naringin to the medium. 3. Hydrolysis of naringin with approporiate concentration of naringinase was carried out linerly up to 80% on the 0.1% substrate solution. 4. The optimum temperature for its activity was $50^{\circ}C$, and this enzyme was inactivated 80% of its total activity at $70^{\circ}C$ for 10 minutes, 40% at $60^{\circ}C$ for 30 minutes. But signifiant decrease of activity were not occurred by heat treatment at $50^{\circ}C$ for 2 hours. 5. Crude enzyme of the naringinase obtained from S-1 strain was competitively inhibited by addition of glucose on the substrate, and inhibitor constant of the glucose on the this enzyme was 1.5 Mol, and inhibition rate were linearly increased according to the increase of sucrose concentration and 56% of its total activity was inhibited at 1 Mol sucrose solution.

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Metabolism of $C^{14}-1-glucose$ and $C^{14}-6-glucose$ by the Ehrlich Ascites Turner Tissue (에르릿히 복수종양의 $C^{14}-1-$ 포도당 및 $C^{14}-6-$포도당 대사에 관한 연구)

  • Kwon, Chang-Rak
    • The Korean Journal of Physiology
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    • v.1 no.1
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    • pp.33-41
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    • 1967
  • The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.

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