• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.292-300
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• 2012

We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.343-351
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• 2012

In this paper, the kinetics of a cloned special glucosidase, named ginsenosidase type III hydrolyzing 3-O-glucoside of multi-protopanaxadiol (PPD)-type ginsenosides, were investigated. The gene (bgpA) encoding this enzyme was cloned from a Terrabacter ginsenosidimutans strain and then expressed in E. coli cells. Ginsenosidase type III was able to hydrolyze 3-O-glucoside of multi-PPD-type ginsenosides. For instance, it was able to hydrolyze the 3-O-${\beta}$-D-(1${\rightarrow}$2)-glucopyranosyl of Rb1 to gypenoside XVII, and then to further hydrolyze the 3-O-${\beta}$-D-glucopyranosyl of gypenoside XVII to gypenoside LXXV. Similarly, the enzyme could hydrolyze the glucopyranosyls linked to the 3-O-position of Rb2, Rc, Rd, Rb3, and Rg3. With a larger enzyme reaction $K_m$ value, there was a slower enzyme reaction speed; and the larger the enzyme reaction $V_{max}$ value, the faster the enzyme reaction speed was. The $K_m$ values from small to large were 3.85 mM for Rc, 4.08 mM for Rb1, 8.85 mM for Rb3, 9.09 mM for Rb2, 9.70 mM for Rg3(S), 11.4 mM for Rd and 12.9 mM for F2; and $V_{max}$ value from large to small was 23.2 mM/h for Rc, 16.6 mM/h for Rb1, 14.6 mM/h for Rb3, 14.3 mM/h for Rb2, 1.81mM/h for Rg3(S), 1.40 mM/h for Rd, and 0.41 mM/h for F2. According to the $V_{max}$ and $K_m$ values of the ginsenosidase type III, the hydrolysis speed of these substrates by the enzyme was Rc>Rb1>Rb3>Rb2>Rg3(S)>Rd>F2 in order.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.5
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• pp.535-542
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• 2006

In this study, removals of 1-naphthol by oxidative-coupling reaction using birnessite, one of natural Mn oxides present in soil, was investigated in various experimental conditions(reaction time, Mn oxide loadings, pH, etc). Removal efficiency of 1-naphthol by birnessite was high in all the experimental conditions, and UV-vis. and mass spectrometric analyses on the supernatant after reaction confirmed that the reaction products were oligomers formed by oxidative-coupling reaction. Pseudo-first order rate constants, f, for the oxidative transformation of 1-naphthol by birnessite was derived from the kinetic experiments under various amount of birnessite loadings, and using the observed pseudo-first order rate constants with respect to birnessite loadings, surface area-normalized specific rate constant, $k_{surf}$ was also determined to be $9.31{\times}10^{-4}(L/m^2{\cdot}min)$ for 1-naphthol. In addition, the oxidative transformation of 1-naphthol was found to be dependent on solution pH, and the pseudo-first order rate constants were increased from 0.129 at pH 10 to 0.187 at pH 4.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1720-1728
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• 2020

We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

• Journal of Korean Society of Environmental Engineers
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• v.35 no.3
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• pp.220-225
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• 2013

The adsorption characteristics of phenol by the powdered activated carbon (PAC) were investigated by series of batch experiments. The pseudo-second-order model described the adsorption kinetics adequately with correlation coefficients over 0.999, indicating chemical adsorption as the rate-limiting step. The kinetic rate constants were from 0.55 to 19.81 mg $mg^{-1}min^{-1}$. The adsorption isotherm followed the Langmuir isotherm, indicating the homogeneous mono-layer adsorption onto the surface of the adsorbent. The values of activation energy, enthalpy and entropy were 17.44 kJ $mol^{-1}$, -8.26 kJ $mol^{-1}$ and -18.94 J $mol^{-1}K^{-1}$, respectively. The Gibbs free energy was in the range of -2.89~-2.14 kJ $mol^{-1}$. The results show that the phenol adsorption is physical, spontaneous and exothermic reaction.

• Journal of Korean Society of Environmental Engineers
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• v.39 no.1
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• pp.26-33
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• 2017

The objective of this study was to evaluate the characteristic of adsorption by using a meso-porous adsorbent (MPA), and investigate the removal efficiency of geosmin which taste and odor causing compounds in drinking water supplies through batch test. The results for the adsorption isotherm was analyzed by using the Langmuir equation and Freundlich equation, generally being applied. And the study showed that the both Langmuir and Freundlich equation explains the results better. Both of pseudo-first-order model and pseudo-second-order model were respectively applied for evaluation of kinetic sorption property of geosmin onto MPA. The adsorption experiment results using MPA showed that maximum adsorption capacity of MPA was lower 7 times than that of GAC, and adsorption rate of MPA was faster 11 times than that of GAC, on the basis of pseudo-first-order model. Therefore, it was determined that MPA was effectively able to remove geosmin in drinking water supplies in short EBCT condition, but regeneration cycle in MAP process was shorter than that in conventional process.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.8
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• pp.1390-1393
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• 2003

Measuring in vivo rate of bone collagen synthesis has so far been technically difficult and often subject to quite large errors. In the present study, bone collagen synthesis rate was measured using a precursor-product method, based on the exchange of $^2$$H_2O$ into amino acids. Mass isotopomer abundance in hydroxyproline from bone collagen was analyzed by gas chromatography/mass spectrometry. The $^2$$H_2O$ labeling protocol consisted of an initial intraperitoneal injection of 99.9% $^2$$H_2O$, to achieve approximately 2.5% body water enrichment followed by administration of 4% $^2$$H_2O$ in drinking water for 9 weeks. Body $^2$$H_2O$ enrichments were stable at 2.7 ∼ 3.0% over labeling Period. In growing rats, the fractional synthesis rate ( $k_{s}$) of bone collagen was 0.066 $\pm$ 0.049 w $k^{-1}$ . The unique features of stable $^2$$H_2O$ pools and label incorporation allowed the precursor-product approach to be used for measuring bone collagen synthesis rate..

• Journal of the Society of Naval Architects of Korea
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• v.39 no.3
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• pp.48-56
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• 2002

The flow characteristics around KRISO 3600TEU container ship model have been experimentally investigated in a circulating water channel. The instantaneous velocity vectors were measured using 2-frame PIV measurement system. The mean velocity fields and turbulent statistics including turbulent kinetic energy and vorticity were obtained by ensemble-averaging 400 instantaneous velocity fields. The free stream velocity was fixed at 0.6m/s and the corresponding Reynolds number was $9{\times}10^5$. The test sections were divided into two regions, three transverse sections of the wake region(Station -0.5767, -1, -3) and five longitudinal sections of the wake((Z/(B/2)=0, 0.1, 0.2, 0.4, 0.6). In the wake region, large-scale longitudinal vortices of nearly same strength are symmetric with respect to the wake centerline and a relatively weak secondary vortex is formed near the waterline. With going downstream, the strength of longitudinal vortex is decreased and the wake region expands.

• Journal of the Institute of Electronics Engineers of Korea SC
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• v.49 no.1
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• pp.47-56
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• 2012

Bicycle has a large amount of kinetic energy available for energy harvesting technology in its speedy and balanced riding movement. Systematic and realistic analysis of its dynamic property is essential to improve the efficiency of energy harvester. However, there has not been enough researches about precise measurement or analysis of bicycle dynamics on real roads. This study aims to investigate the characteristics of vibrational movement of bicycle using MEMS-based accelerometer and to develop a prototype of electromagnetic energy harvester with nonlinear behavior which is proper to the random vibrations accompanied in bicycle riding. The vibrational components have average magnitude of 1 g and turn out to be independent of riding speed. The developed prototype of energy harvester was installed on a front port of a bicycle to use this ambient vibration and generated an average electrical power of 1.5 mW which is enough to support power for most of portable sensors and short range radio-frequency communication. Further study about isolation of vibration from a rider and conversion efficiency is ongoing. The developed energy harvester is expected to be a platform technology for sustainable portable power supply for various smart IT devices and applications.

• Journal of the Korean Chemical Society
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• v.43 no.4
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• pp.461-468
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• 1999

Acetolactate synthase (ALS) catalyzes the first common reaction in the biosynthesis of branched-chain amino acids, valine, leucine, and isoleucine. ALS is the common target of several classes of structurally diverse herbicides, the triazolopyrimidines, the imidazolinones, the sulfonylureas, and pyrimidyl-oxy-benzoates. We examined ihibitory activities of newly synthesized triazolopyrimidine sulfonamide derivatives using partially purified ALS from barley. $IC_{50}$ values for the active derivatives are 0.5nM∼8$\mu$M. Among them TP1 and TP2 are the most potent ALS inhibitors with $IC_{50}$ values of 0.5nM and 1.6nM, respectively. These inhibitors are more potent in the inhibition of barley ALS than commercial herbicides, Metosulam ($IC_{50}$;3.6 nM), Flumetsulam ($IC_{50}$;126 nM), and Cadre ($IC_{50};4 {\mu}M$). The progress curves for inhibition of ALS by TP2 showed that the amount of inhibition increases with time. The inhibition of ALS by TP2 was mixed-type inhibition with respect to pyruvate. Dual inhibition analyses of TP2 versus an imidazolinone, Cadre, and Leu showed parallel and intercepting kinetic pattern, respectively. The results suggest that TP2 binds to ALS competively with Cadre but not with Leu. Chemical modification of cysteinly residues in ALS decreased the sensitivity of ALS to Leu, while the modification did not affect the sensitivity of ALS to TP2 and Cadre.