A gene encoding ${\beta}$-glucosidase was cloned from Weissella cibaria 37, an isolate from human feces. Sequence analysis showed that the gene could encode a protein of 415 amino acids in length, and the translated amino acid sequence showed homology (34-31%) with glycosyl hydrolase family 1 ${\beta}$-glucosidases. The gene was overexpressed in E. coli BL21(DE3) using pET26b(+) and a 50 kDa protein was overproduced, which matched well with the calculated size of the enzyme, 49,950.87 Da. Recombinant ${\beta}$-glucosidase was purified by using a his-tag affinity column. The purified ${\beta}$-glucosidase had an optimum pH and a temperature of 5.5 and $45^{\circ}C$, respectively. Among the metal ions (5mM concentration), $Ca^{2+}$ slightly increased the activity (108.2%) whereas $Cu^{2+}$ (46.1%) and $Zn^{2+}$ (56.7%) reduced the activity. Among the enzyme inhibitors (1 mM concentration), SDS was the strongest inhibitor (16.9%), followed by pepstatin A (45.2%). The $K_m$ and $V_{max}$ values of purified enzyme were 4.04 mM and 0.92 ${\mu}mol/min$, respectively, when assayed using pNPG (p-nitrophenyl-${\beta}$-D-glucopyranoside) as the substrate. The enzyme liberated reducing sugars from carboxymethyl cellulose (CMC).
Gyejibokryeong-hwan (GJBRH) has been used for treatment of patients with climacteric syndrome. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer (UPLC-ESI-MS) method was established for the simultaneous quantification of seven marker compounds in GJBRH extract. In addition, we assessed the antioxidant effects of GJBRH. All analytes were separated by gradient elution using two mobile phases on a UPLC BEH $C_{18}$ column and maintained at $45^{\circ}C$. The antioxidant activities of GJBRH were evaluated by measuring free radical scavenging activities on 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS) and relative electrophoretic mobility (REM). Regression equations of the seven compounds were acquired with $r^2$ values ${\geq}0.9988$. The amounts of the seven compounds, amygdalin, albiflorin, paeoniflorin, coumarin, cinnamic acid, cinnamaldehyde, and paeonol in GJBRH water extract were 21.71, 2.16, 17.17, 1.97, 0.40, 0.78, and 3.42 mg/g, respectively. The GJBRH showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were $54.18{\mu}g/mL$ and $79.53{\mu}g/mL$. Furthermore, GJBRH reduced the oxidation properties of LDL induced by $CuSO_4$.
A thiol protease has been isolated and partially purified from encysted brine shrimp Artemia franciscana using a four-step procedure(filtration, salting out, gel filtration and ion exchange chromatography). The optimum pH of the enzyme for caseinolytic activity was appeared to be 3.0, and the enzymematic activity was stable up to pH 6.0 but lost completely at the pH higher than 8.0. The optimal temperature of the enzyme was appeared to be 35$^{\circ}$C, and ninety percent of the enzyme activity was lost at 45$^{\circ}$C. Various metal ions, e.g., zinc, copper, iron, inhibited the enzyme activity; however, heavy metal chelator, e.g., EDTA, stimulated the enzyme activity. The protease was concluded to be a member of the thiol group protease, since it was inhibited by thiol protease inhibitors and iodoacetate. The protease was also concluded to be a acid protease based on optimum pH.
This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.
Water repellency which affects infiltration, evaporation, erosion and other water transfer mechanisms through soil has been observed under several natural conditions. Water repellency is thought to be caused by hydrophobic organic compounds, which are present as coatings on soil particles or as an interstitial matter between soil particles. This study was conducted to evaluate the characteristics of the water repellent soil and transport characteristics of trace elements within this soil. Capillary height of the water repellent soil was measured. Batch and column studies were accompanied to identify sorption and transport mechanism of trace elements such as $Cu^{2+}$, $Mn^{2+}$, $Fe^{2+}$, $Zn^{2+}$ and $Mo^{5+}$. Difference of sorption capacity between common and repellent soils was observed depended on the degree of repellency. In the column study, the desorption of trace elements and the spatial concentration distribution as a function of time were evaluated. The capillary height was in the repellency order of 0% > 15% > 40% > 70% > 100%. No water was absorbed in soil indicating >70% repellency. Using trace elements, $Fe^{2+}$ and $Mo^{5+}$ showed higher sorption capacity in the repellent soil than in non-repellent soil. The sorption performance of $Fe^{2+}$ was found to be in the repellency order of 40% > 15% > 0%. Our results found that transfer of $Mo^{5+}$ had similar sorption tendency in soils having 0%, 15% and 40% repellency at the beginning, however, the higher desorption capacity was observed as time passes in the repellent soil compared to in non-repellent soils.
Kim, Tae-Gyun;Cho, Kang-Woo;Song, Kyung-Guen;Yoon, Min-Hyuk;Ahn, Kyu-Hong;Hong, Sung-Kwan
Journal of Korean Society of Environmental Engineers
/
v.31
no.7
/
pp.483-490
/
2009
The aims of this study were to evaluate the removal efficiency for various pollutants in urban storm runoff by a filtration device, and to determine design parameters depending on filter media properties. Appropriate selection of filter media will affect the size and life time of the filtration device. Sets of column tests were performed in order to evaluate the removal efficiency by perlite and a synthetic resin. An investigation of surface properties including CEC (cation exchange capacity) and zeta-potential suggested that the perlite had a superior adsorption capability for cationic pollutants. TCODcr and turbidity were analyzed to investigate the removal characteristic of particulate pollutant. In both columns, the particles in the collected storm runoff was almost completely capture with a small EBCT (empty bed contact time) of 2.5 minutes. Complete clogging at the EBCT of 2.5 minutes occurred after 630 minutes in the perlite column and 810 minutes in the resin column. The removal efficiency of TCODcr and turbidity at the EBCT of 2.5 minutes decreased to below 70% due to an wall effect. The removal efficiency for dissolved pollutant (SCODcr) was negligible due to the insufficient contact time for adsorption. The removal of heavy metals (Cu, Zn, Pb) was mostly ascribed to the filtration of particles containing metals, since the relationship between CEC and the removal efficiency was not apparent. The result of this study would be valuable for the application of filtration device to control of urban storm runoff.
In this study, coal combustion ash (CCA) was evaluated for its stabilization effect on acidic mine waste with column experiment. Total of six treatments were installed depending on mixing ratio between coal wastes and CCA (0, 20, 40%) and mixing method (completely mixing and layered). Artificial acidic rain (pH 5.6) was used for feeding solution with flow rate of $0.05mL\;min^{-1}$. Result showed that higher pH of leachate was observed as more CCA was mixed. The highest pH in leachate was measured when 40% of CCA was mixed with coal waste (pH of 5.8). Also, complete mixing with CCA and coal waste was more effective to increase the pH of leachate than layered treatment. Regarding the reduction of soluble Fe amount, the highest efficiency (78%) was observed when 20% of coal ash was completely mixed with mine waste. Based on those result, optimum mixing ratio of coal ash with mine waste can be ranged 20-40% depending on environmental circumstances in the field.
Kim, Kyeong-Hong;Lee, Jae-Hak;Shin, Kyung-Soon;Pae, Se-Jin;Yoo, Sin-Jae;Chung, Chang-Soo;Hyun, Jung-Ho
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
/
v.5
no.3
/
pp.224-232
/
2000
Inorganic nutrient concentrations in relation to springtime physical parameters of the Yellow Sea were investigated during April 1996. Three major water masses, i.e., the Yellow Sea Warm Current Water (YSWC), Coastal Current Water (CCW) and Changjiang River Diluted Water (CRDW), prevailed in the study area. Water masses were vertically wel1 mixed throughout the study area, and nutrients were supplied adequately from bottom to surface layer. As result of ample nutrients supplied by vertical mixing together with progressed daylight condition, springtime phytoplankton blooms were observed, which was responsible for the depletion of inorganic nutrients in surface water column. Low nutrients concentration in bottom water of the central Yellow Sea (Stn. D9; nitrate: <2 ${\mu}$M, phosphate: <0.3 ${\mu}$) was associated with the entrance of YSWC which is characterized by high temperature and salinity. Influenced by runoff and vertical tidal mixing, CCW with high nutrient concentrations probably associated with China and Korea coastal waters with high nutrients concentration. For the local scale of inorganic nutrient distribution, nutrient transfers from coast to central areas were limited due to restriction imposed by tidal fronts (Stn. D6) and thus affected the horizontal nutrient profiles. Relatively high phytoplankton biomass was observed in the tidal front (Chl-${\alpha}$=12.38 ${\mu}$gL$^{-1}$) during the study period. Overall, the springtime nutrient distribution patterns in the Yellow Sea appeared to be affected by: (1) Large-scale influx of YSWC with low nutrient concentrations and CCW with high nutrient concentrations influenced by Korea and China coastal waters; (2) vertical mixing of water mass and phytoplankton distribution; and (3) local-scale tidal front as well as phytoplankton blooms alongthe tidal front.
Pollutants from industry, mining, agriculture, and other sources have contaminated sediments in many surface water bodies. Sediment contamination poses a severe threat to human health and environment because many toxic contaminants that are barely detectable in the water column can accumulate in sediments at much higher levels. The purpose of this study was to make optimal treatment and disposal plan o( sediment for water quality improvement in small-scale resevoir based on an evaluation of degree of contamination. The degree of contamination were investigated for 23 samples of 9 site at different depth of sediment in small-scale J river. Results for analysis of contaminated sediments were observed that copper concentration of 4 samples were higher than the regulation of hazardous waste (3 mg/L) and that of all samples were exceeded soil pollution warning levels for agricultural areas. Lead and mercury concentration of all samples were detected below both regulations. Necessary of sediment dredge was evaluated for organic matter and nutrient through standard levels of Paldang lake and the lower Han river in Korea and Tokyo bay and Yokohama bay in Japan. The degree of contamination for organic matter and nutrient was not serious. Compared standard levels of Japan, America, and Canada for heavy metal, contaminated sediment was concluded as lowest effect level or limit of tolerance level because standard levels of America and Canada was established worst effect of benthic organisms. The optimal treatment method of sediment contained heavy metal was cement-based solidification/stabilization to prevent heavy metal leaching.
Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.
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