• Title/Summary/Keyword: 기질 친화상수

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An Effect of Ethanol on Polypyrrole-Glucose Oxidase Enzyme Electrode (Polypyrrole-Glucose oxidase 효소전극의 Ethanol 첨가효과)

  • 김현철;구할본;사공건
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1999.11a
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    • pp.147-150
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    • 1999
  • In the case of immobilizing of glucose oxidase in organic polymer using electrosynthesis, the glucose oxidase obstructs charge transfer and mass transport during the film growth. This may lead to short chained polymer and/or make charge-coupling weak between the glucose oxidase and the backbone of the polymer. That is mainly due to insulating property and net chain of the glucose oxidase. Since being the case, it is useless to increase in amount of glucose oxidase more than reasonable in the synthetic solution. We establish qualitatively that amount of immobilization can be improved by adding a little ethanol in the synthetic solution. As ethanol was added by 0.1 rnol dm" in the synthetic solution, Michaelis-Menten constants of the resulting enzyme electrode decreased from 30.7 mmol $dm^{-3}$ to about 2 mmol $dm^{-3}$. That suggests increase in affinity of the enzyme electrode for glucose and in amount of the immobilized enzyme.zyme.

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Substrate Variety of a Non-metal Dependent Tagatose-6-phosphate Isomerase from Staphylococcus aureus (Staphylococcus aureus 유래 비금속성 이성화효소인 Tagatose-6-phosphate Isomerase의 기질다양성)

  • Oh Deok-Kun;Ji Eun-Soo;Kwon Young-Deok;Kim Hye-Jung;Kim Pil
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.106-111
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    • 2005
  • To investigate the substrate variety of a putative non-metal dependent isomerase, the tagatose-6-phosphate isomerase (E.C. 5.3.1.26) structural genes (lacB; 510bp and lacA; 430bp) of Staphylococcus aureus were subcloned and co-expressed. Based on the substrate configuration, various aldoses were surveyed for substrate of ketose isomerization. Among the 10 aldoses tested, D-ribose and D-allose were isomerized by the enzyme. The subunit A and B showed more than $95\%$ activity for D-ribose and $75\%$ for D-allose in the presence of 1mM EDTA compared with non-EDTA conditions, which implying tagatose-6-phosphate isomerase is a non-metal dependent isomerase. Each of subunit A or subunit B alone showed no activity for any of the substrates tested. The affinity constant ($K_m$) of tagatose-6-phosphate isomerase against D-ribose and D-allose were 26 mM and 142 mM, respectively.

The Study of TCE Dechlorination using Geobacter lovleyi with Slow Release Substrate Applied (Slow Release Substrate를 이용한 Geobacter lovleyi의 TCE 탈염소화 연구)

  • Cha, Jae Hun;An, Sang Woo;Park, Jae Woo;Chang, Soon Woong
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.9
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    • pp.53-59
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    • 2012
  • This study investigated characteristics of decomposition of tetrabutoxysilane (TBOS) as a slow release substrate (SRS) and on effect of TBOS decompostion compounds (acetate and butylate) for anaerobic dechlorination of trichloroethylene (TCE). In the batch experiment, TCE, cis-dichloroethene (cis-DCE), 1-butanol and TBOS were analysed by GC/FID and acetate and butylate were measured by HPLC. 1M of TBOS transferred and accumulated 4M of 1-butanol by abiotically hydrolysis reaction. The hydrolysis rate was in a range of 0.186 ${\mu}M/day$. On other hand, 1-butanol fermented to butyrate and acetate with indigenous culture from natural sediments. This results showed that TBOS could be used a slow release substrate in the natural sites. The dechlorinated potential of TCE with acetate and butyrate was increased with a decreasing initial TCE concentrations. In addition, first order coefficients of dechlorination with acetate as electron donor was higher then that with butyrate. It is because that dechlorination of Geobacter lovleyi was affected by substrate affinity, biodegradability and microbial acclimation on various substrates. However, dechlorinated potential of Geobacter lovleyi was decreased with accumulation cis-DCE in the anaerobic decholoronation process. The overall results indicated that SRS with Geobacter lovleyi might be a promising material for enhancing dechlorination of TCE on natural site and cis-DCE should be treated by ZVI as reductive material or by coexisting other dechlorinated bacteria.

Production of Chitosna Oligosaccharides Using Chitin-Immobilized Enzyme (키틴 고정화 효소를 이용한 키토산 올리고당의 생산)

  • 전유진;박표잠;변희국;송병권;김세권
    • KSBB Journal
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    • v.13 no.2
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    • pp.147-154
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    • 1998
  • Enzymatic hydrolysis using an immobilized enzyme was carried out to produce chitosan oligosaccharides (COSs) from chitosan effectively. Chitosanase was immobilized on eight different carriers by physical adsorption. The enzyme immobilized on chitin had higher activity than those immobilized on the other carriers in spite of its lower adsorption. The activity of chitin-immobilized enzyme was more than 90% of the original activity. Optimal temperature of the immobilized enzyme increased by about $15^{\circ}C$ and its thermostability was excellent in relatively wide range of temperature. But its effects of pH did not improve compared to the free enzyme. The immobilized enzyme produced 153 mg/g chitosan of the reducing sugar for 3hrs of hydrolytic incubation time. The total content of higher oligomers, tetramer to hexamer, among amount of total COSs obtained for 2hrs was more than 90%. In kinetic parameters for both enzymes, immobilized enzyme showed lower affinity for substrate and reaction rate than free enzyme, however, no reduction of the rate for high substrate concentrations. Consequently, chitin-immobilized could effectively hydrolyse chitosan and produce the higher COSs without activity decrease in comparison with the free enzyme.

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Purification and Characterization of $\beta$-Galactosidase from Sea Urchin, Hemicentrotus pulcherrimus (성게로부터 분리한 $\beta$-galactosidase의 정제 및 특성)

  • KIM Gyu-Hyung;KIM Yong-Tae;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.5
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    • pp.637-644
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    • 1998
  • [ $\beta$ ]-Galactosidase was extracted from the internal organ of sea urchin, Hemicentrotus pulcherrimus The enzyme was purified 384.6-fold over the crude extract by the sequential chromatographic methods including DEAE-Sephadex A-25, CM-Cellulose, and Con A-Sepharose 4B affinity chromatography with a recovery $1.26\%$. The molecular weight of the purified enzyme was estimated approximately 94 kDa as monomeric term by SDS-PAGE and Sephadex G-150 gel chromatography. The maximum enzymatic activity was observed at pH 3.0 and $50^{\circ}C$ but the one was stable over the ph range or 3.0$\~$5.0 and below $37^{\circ}C$. The $K_m$ and $V_{max}$ values against PNPG (P-nitrophenyl $\beta$-D-galactopyranoside) were 15.0 mM and 214 $\mu$mole/min per mg protein, respectively. The enzymatic activity was activated by $Ba^{2+}$, but significantly inhibited by $DEP,\;Hg^{2+},\;Sn^{2+}$ and galactose.

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Studies on the processing of rapid fermented anchovy prepared with low salt contents by adapted microorganism. -2. Thermodynamic characteristics of microbial extracellular protease isolated from fermented fish paste- (미생물을 이용한 저식염 멸치젓의 속성발효에 관한 연구 -2. 젓갈에서 분리한 단백질분해효소의 열역학적 특성-)

  • Cha, Yong-Jun;Lee, Eung-Ho
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.325-329
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    • 1990
  • This study was undertaken to determine thermodynamic characteristics of B. subtilis p-4 and B. licheniformis p-5 proteases isolated from fermented anchovy paste. $K_m$ values of two proteases for casein as a substrate were 0.38mM in p-4 protease and 0.18mM in p-5 protease, respectively. Denaturation constants($K_D$) of p-4 and p-5 proteases were $12.2{\times}10^{-5}/sec\;and\;19.0{\times}10^{-5}/sec\;at\;40^{\circ}C,\;and\;35.7{\times}10^{-5}/sec\;and\;46.3{\times}10^{-5}/sec\;at\;50^{\circ}C$, respectively. Activation energies($E_a$) of p-4 and p-S pmteases were 19.6 Kcal/mole and 15.2kcal/mole, respectively. Free energy of activation(${\Delta}G^*$), activation enthalpy(${\Delta}H^*$) and activation entropy(${\Delta}S^*$) at $40^{\circ}C$ were 23.21Kcal/mole, 18.98Kcal/mole and -13.50 eu, respectively for p-4 protease and 22.93Kcal/mo1e, 14.58Kcal/mole and -26.68 eu, respectively for p-5 protease. The major amino acids in p-4 protease(151 residues of amino acid) were Gly, Glu, Pro, Asp, Ser, Ala, Lys and Leu, while those in p-5 protease(247 residues of amino acid) were Gly, Glu, Asp, Ala and Leu. It may be concluded that heat denaturation of two proteases showed liner regression curve and p-5 protease was more sensitive to heat than p-4 protease.

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Evaluation of Kinetic Constant and Effect of Effluent Recycling in Wastewater Treatment from Fisheries Processing Plant using EMMC Process (EMMC공정을 이용한 수산물 가공공장 폐수처리에서 동력학적 인자 평가와 유출수반송의 영향)

  • Jeong, Byung-Gon
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.12 no.1
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    • pp.1-8
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    • 2009
  • EMMC(Entrapped Mixed Microbial Cell) process which is a kind of active cell immobilizing method was applied to treat fisheries processing wastewater biologically. Kinetic constants were calculated for organic and nitrogen removal and effect of effluent recycling on system performance was evaluated also. Yield coefficient, Y showed relatively low value compared with Y value obtained from conventional activated sludge process. It means that EMMC process can reduce amount of excess sludge significantly compared with conventional activated sludge process. Endogenous respiration coefficient $k_e$ of EMMC process also showed relatively low value compared with that of conventional activated sludge process. Yield coefficient Y, endogenous respiration coefficient $k_e$ and half saturation constant $k_s$ obtained from EMMC process in terms of nitrification were compared with reported value from literature based on suspended growth nitrification system. The value of Y obtained from this study has no difference compared with values obtained from literature review and $k_e$ of this study was low but $k_s$ of this study was high compared than values obtained from suspended growth nitrification system. To evaluate the effect of internal recycling on system performance, system was operated with internal recycling ratio of 1.5Q, 2.0Q, 2.5Q and 3.0Q. increase of internal recycling ratio effect more greatly on improvement of denitrification efficiency than that of nitrification efficiency. Accordingly, optimization of internal recycling ratio has to be based on improvement of anoxic reactor performance.

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Characterization of Carboxymethyl Cellulase Produced by Cellulomonas sp. CS1-1 on Microcrystalline Cellulose (Cellulomonas sp. CS1-1이 미소결정성 섬유소로부터 생산한 Carboxymethyl Cellulase의 효소적 성질)

  • Park, Jong-Soo;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.24 no.2
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    • pp.275-282
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    • 1997
  • The prcx.iuction of extracellular 1,4-${\beta}$-glucanase by Cellulomonas sp. CS1-1 on microcrystalline cellulose, sigmacell was maximal after 5-day cultivation as 280 units/mL, which was three times higher than the level produced on carboxymethyl cellulose. A carboxymethyl cellulase containing the carbohydrate of 8.2% was purified from the culture filtrate by successive procedures of column chromatographies. Purification factor was calculated as 22-folds with the specific carboxymethyl cellulase activity of 31.9 units/mg. The molecular weight and isoelectric point of the purified enzyme were 54,000 and pI 5.4, respectively. The optimal pH and temperature were 6.0 and $45^{\circ}C$, and the enzyme was stable between pH 6.5 and 7.5 and below $50^{\circ}C$. The estimated Km and Vmax were 10 mg/mL and $6.25{\mu}mol/min$ for carboxymethyl cellulose and 30.3 mg/mL and $2.85{\mu}mol/min$ for sigmacell, respectively. The enzyme was partially inhibited by $Ag^+$, $Zn^{+{+}}$, $Fe^{+{+}}$ and EDTA, while completely inhibited by $Cd^{+{+}}$ and $Hg^{+{+}}$ at 1 mM concentration.

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Growth kinetics and chlorine resistance of heterotrophic bacteria isolated from young biofilms formed on a model drinking water distribution system (모델 상수관망에 형성된 초기 생물막에서 분리한 종속영양세균의 생장 동역학 및 염소 내성)

  • Park, Se-Keun;Kim, Yeong-Kwan;Oh, Young-Sook;Choi, Sung-Chan
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.355-363
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    • 2015
  • The present work quantified the growth of young biofilm in a model distribution system that was fed with chlorinated drinking water at a hydraulic retention time of 2 h. Bacterial biofilms grew on the surface of polyvinyl chloride (PVC) slides at a specific growth rate of $0.14{\pm}0.09day^{-1}$ for total bacteria and $0.16{\pm}0.08day^{-1}$ for heterotrophic bacteria, reaching $3.1{\times}10^4cells/cm^2$ and $6.6{\times}10^3CFU/cm^2$ after 10 days, respectively. The specific growth rates of biofilm-forming bacteria were found to be much higher than those of bulk-phase bacteria, suggesting that biofilm bacteria account for a major part of the bacterial production in this model system. Biofilm isolates exhibited characteristic kinetic properties, as determined by ${\mu}_{max}$ and $K_S$ values using the Monod model, in a defined growth medium containing various amounts of acetate. The lowest ${\mu}_{max}$ value was observed in bacterial species belonging to the genus Methylobacterium, and their slow growth seemed to confer high resistance to chlorine treatment (0.5 mg/L for 10 min). $K_S$ values (inversely related to substrate affinity) of Sphingomonas were two orders of magnitude lower for acetate carbon than those of other isolates. The Sphingomonas isolates may have obligate-oligotrophic characteristics, since the lower $K_S$ values allow them to thrive under nutrient-deficient conditions. These results provide a better understanding and control of multi-species bacterial biofilms that develop within days in a drinking water distribution system.

Biodeodorization of Trimethylamine by Biofilter Packed with Waste Tire-Chips (폐타이어칩 충진형 바이오 필터에 의한 Trimethylamine 제거)

  • Park, Hun-Ju;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.8
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    • pp.789-797
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    • 2008
  • This study was conducted to investigate removal characteristics of gaseous trimethylamine(TMA) through biofilter packed with waste tire-chips. The sludge in this experiment was collected from an activated sludge operated in a wastewater treatment facility treating malodorous pollutants. The nominal amount of collected sludge was inoculated through packing materials in the filter. The removal efficiencies for varying concentrations and SVs(Space velocity) were assessed based on TMA, COD$_{Cr}$, NO$_3{^-}$-N, NO$_2{^-}$-N, NH$_4{^+}$-N and EPS(Extracellular Polymeric Substances) in leachate, since biofilter had been steady-stately operated. The influent concentration of 10 ppm of TMA was removed to approximately 95% regardless of changing SV at 120 and 180 hr$^{-1}$, but it was lowered to 80 to 90% at SV 240 hr$^{-1}$. As influent concentration was gradually increased from 5 to 55 ppm, the removal efficiencies of TMA were initially high for 95% in the range of 5 to 10 ppm, but lowered to 80% for 10 to 30 ppm. As a part of kinetic study for TMA decomposition, V$_m$(maximum substrate removal rate) and $K_s$(substrate infinity coefficient) were 14.3 g$\cdot$m$^{-3}$$\cdot$h$^{-1}$ and 0.043 g$\cdot$m$^{-3}$, respectively while adapted period was shown in the range of 100 to 150 hr. Also, the EPS concentration was consistently observed from the leachate showing 100 to 200 ppm, which indicates that biofilm has been continuously formed and sustained throughout tire-chips packed reactor.