• Microbiology and Biotechnology Letters
• /
• v.47 no.2
• /
• pp.259-268
• /
• 2019

In this study, the extracellular metalloprotease from Serratia marcescens PPB-26 was purified to homogeneity via ethanol fractionation and DEAE-cellulose column chromatography. Thus, a 3.8-fold purification was achieved with a 20% yield and specific activity of 76.2 U/mg. The purified protease was a 50-kDa monomer whose optimum pH and temperature for activity were 7.5 and $30^{\circ}C$ respectively; however, it was found to remain active in the 5-9 pH range and up to $40^{\circ}C$ for 6 h. The protease had a half-life of 15 days at $4^{\circ}C$, an optimum reaction time of 10 min, and an optimum substrate (casein) concentration of 0.25%. Furthermore, the Michaelis constant ($K_m$) and reaction velocity ($V_{max}$) of the protease were calculated to be 0.28% and $111.11{\mu}moles/(min{\cdot}mg)^{-1}$, respectively. The protease was stable when subjected to metal ions (2 mM), showing increased activity with most (especially $CoCl_2$ and $MgSO_4$ (30.54% increase)). It was also stable when exposed to oxidizing agents, bleaching agents, and detergents (5% v/v for 60 min). It retained 93% of its activity in non-ionic detergents (Tween-20, Tween-80, and Triton X-100). Moreover, wash performance analysis in commercial detergents (Ariel and Tide) showed that not only was the protease capable of protein stain removal, but also reduced cleaning time by 80% when added to detergents. Thus, the Serratia marcescens PPB-26 metalloprotease appears to be a promising new candidate as a laundry additive in the detergent industry.

• Journal of Applied Biological Chemistry
• /
• v.44 no.1
• /
• pp.16-19
• /
• 2001

Soybean plants(Glycine max [L.] merr.) inoculated with Bradyrhizobium japonicum MN110 were grown in growth chambers under 400 or $800{\mu}l{\cdot}l^{-1}$ atmospheric $CO_2$ and harvested at 25, 28, 32, and 35 DAT to examine the effect of $CO_2$ enrichment on phosphorus accumulation, uptake, and utilization efficiency during vegetative growth. Phosphorus concentration in leaf was lower in high $CO_2$ plant by 47% at 25 DAT and 34% at 35 DAT than those in the control plant but phosphorus concentrations in stem, root and nodule were not affected by $CO_2$ enrichment. Total phosphorus accumulation increased 3.9-fold in high $CO_2$ plant and 3.2-fold in the control plant between 25 and 35 DAT. Elevated $CO_2$ caused a decrease in the whole plant phosphorus concentration by 35%, which was due almost entirely to a decrease in the phosphorus concentration of leaves. $CO_2$ enrichment increased phosphorus utilization efficiency in the whole plant by 70% during the experimental period. Plants exposed to high $CO_2$ had larger root systems than under ambient $CO_2$, but high $CO_2$ plants had lower P-uptake efficiency. Averaged over four harvests, plants at high $CO_2$ had 38% larger root mass that was more than offset the 20% lower efficiency of P-uptake and accounted for increased phosphorus accumulation by high $CO_2$ plant. These results suggest that the reduced phosphorus concentration in soybean plant under $CO_2$ enrichment may be an acclimation response to high $CO_2$ concentration or enhanced starch accumulation, resulting in the plants to have a lower phosphorus requirement on a unit dry weight basis or a high phosphorus utilization efficiency under these conditions.

• BMB Reports
• /
• v.37 no.6
• /
• pp.684-690
• /
• 2004

Heparin lyase I was purified to homogeneity from Bacteroides stercoris HJ-15 isolated from human intestine, by a combination of DEAE-Sepharose, gel-filtration, hydroxyapatite, and CM-Sephadex C-50 column chromatography. This enzyme preferred heparin to heparan sulfate, but was inactive at cleaving acharan sulfate. The apparent molecular mass of heparin lyase I was estimated as 48,000 daltons by SDS-PAGE and its isoelectric point was determined as 9.0 by IEF. The purified enzyme required 500 mM NaCl in the reaction mixture for maximal activity and the optimal activity was obtained at pH 7.0 and $50^{\circ}C$. It was rather stable within the range of 25 to $50^{\circ}C$ but lost activity rapidly above $50^{\circ}C$. The enzyme was activated by $Co^{2+}$ or EDTA and stabilized by dithiothreitol. The kinetic constants, $K_m$ and $V_{max}$ for heparin were $1.3{\times}10^{-5}\;M$ and $8.8\;{\mu}mol/min{\cdot}mg$. The purified heparin lyase I was an eliminase that acted best on porcine intestinal heparin, and to a lesser extent on porcine intestinal mucosa heparan sulfate. It was inactive in the cleavage of N-desulfated heparin and acharan sulfate. In conclusion, heparin lyase I from Bacteroides stercoris was specific to heparin rather than heparan sulfate and its biochemical properties showed a substrate specificity similar to that of Flavobacterial heparin lyase I.

• YAKHAK HOEJI
• /
• v.51 no.2
• /
• pp.133-139
• /
• 2007

A simple and sensitive HPLC-MS method for quantitation of 10${\alpha}$-methoxy-9,10-dihydrolysergol (MDL), the main metabolite of nicergoline, in human plasma was developed and the bioavailability parameters of MDL was assessed in Korean healthy male volunteers. Clomipramine was used as an internal standard. MDL and internal standard in plasma sample were extracted using ethyl acetate. A centrifuged upper layer was then evaporated and reconstituted with mobile phase of 10 mM ammonium acetate-acetonitrile (10 : 90, v/v). The reconstituted samples were injected into a Zorbax SB-C8 column (2.1${\times}$150 mm,5 ${\mu}$m) at a flow-rate of 0.3 ml/min. Using MS with selected ion monitoring (SIM) mode, MDL and clomipramine were detected without severe interference from human plasma matrix. MDL produced a protonated molecular ion ([M+H]$^+$) at m/z 287. Internal standard produced a protonated molecular ion ([M+H]$^+$) at m/z 315. A linear relationship for MDL was found in the range of 2.5${\sim}$100 ng/ml. The lower limit of quantitation (LLOQ) was 2.5 ng/ml with acceptable precision and accuracy. The intra- and inter-day validation for all coefficients of variation (R.S.D.%) were found less than 15%. Main pharmacokinetic parameters of 30 mg of nicergoline were revealed as follows: AUC$_t$ 321.1${\pm}$64.5 ng${\cdot}$hr/ml, C$_{max}$, 51.2${\pm}$25.3 ng/ml, T$_{max}$ 3.6${\pm}$1.5 hr, K$_{el}$ 0.12${\pm}$0.07 hr$^{-1}$ and t$_{1/2}$ 7.6${\pm}$3.4 hr. Inter subject variations and race differences were shown in comparison with the published data in the literature.

• Journal of Pharmaceutical Investigation
• /
• v.22 no.2
• /
• pp.125-131
• /
• 1992

The effects of domperidone, scopolamine butylbromide and cimetidine on the absorption and bioavailability of ciprofloxacin were studied in female rats. Ciprofloxacin was given in a single oral dose of 30 mg/kg to control group. Ciprofloxacin was concurrently administered with domperidone $(T_1\;group)$, scopolamine butylbromide $(T_2\;group)$, and cimetidine $(T_3\;group)$ to rats, respectively. Significantly changed pharmacokinetic parameters observed in $T_2$group when compared with control group were first-order absorption rate constant, $Ka(4.43{\pm}0.85$\;versus\;2.86{\pm}0.41\;hr^{-1},\;p<0.05)$, time needed to reach peak concentration, $T_{max}\;(32.27{\pm}2.46\;versus\;51.75{\pm}5.51\;min,\;p<0.05)$, area under the plasma concentration-time curve, AUC $(332{\pm}19\;versus\;477{\pm}27\;{\mu}g{\cdot}min/ml,\;p<0.05)$ and absolute bioavailability, Fabs $(60.6{\pm}3.6\;versus\;87.0{\pm}5.0%,\;p<0.05)$. On the other hand, domperidone and cimetidine did not significantly affect the absorption of ciprofloxacin. It is suggested that when scopolamine butylbromide is selected for clinical use, there is need for awareness of the reduction in absorption rate and the enhancement in absorption extent of ciprofloxacin.

• Journal of Life Science
• /
• v.17 no.2 s.82
• /
• pp.266-271
• /
• 2007

Bisphenol A (BPA) has been widely used as a monomer for production of epoxy resins and polycarbonate plastics. The annual production of BPA exceeds 640,000 metric tons in worldwide. BPA, a suspected phenolic endocrine disrupter, is moderately soluble and frequently detected in industrial wastewater. To date, HPLC and GC has been used for BPA analysis. However, HPLC and GC-analysis need high operation lost, experts, and an elaborate pre-treatment of samples, and is difficult to apply on-time and mass analysis. Therefore, simple, mass and rapid detection of BPA in environments is necessary. In the present study, spectrophotometric method of BPA quantification was developed. Based on blue-color product formation with BPA and ferric chloride/ferricyanide under the optimized conditions, the standard curve was acquired $({\lambda}_{750}=0.061\;BPA\;[{\mu}M]+0.07155,\;R^2=0.992)$. Using an established method, the BPA contents in the soil extract, and different water samples and living products, including disposable syringe, cup and plastic tube, were analyzed. The results suggested that the method is useful for BPA determination from different massive samples. Since the BPA metabolites, nontoxic 4-hydroxyacetophenone or 4-hydroxybenzaldehyde, did not form blue-color product, this method is also useful to screen a microorganism for BPA bioremediation.

• Korean Journal of Ecology and Environment
• /
• v.39 no.3 s.117
• /
• pp.340-351
• /
• 2006

To reveal physicochemical factors contributing to variation of phytoplankton communities, the study was carried out biweekly at 6 stations from Feb. 2004 to Feb. 2005 in the lower part of the Han River, Korea. As results, water temperature was changed from $0.3^{\circ}C$ to $26.6^{\circ}C$, pH: 6.6${\sim}$9.1, DO: 1.89${\sim}$22.23 mg $L^{-1}$, BOD: 0.38${\sim}$9.20 mg $L^{-1}$, COD: 1.4${\sim}$15.2 mg $L^{-1}$, Conductivity: $62.5{\sim}500.0\;{\mu}s\;cm^{-1}$, SS: 3.00${\sim}$159.3 mg $L^{-1}$, and Chl a $1.7{\sim}71.3\;{\mu}g\;L^{-1}$. Phytoplankton standing crops ranged from min. $3.6{\times}10^2\;cells\;mL^{-1}$ (July 2004, St. 3) to max. $2.3{\times}10^4\;cells\;mL^{-1}$ (Feb. 2005, St. 6), and mean of those varied from $5.9{\times}10^3\;cells\;mL^{-1}$in spring, $2.1{\times}10^3\;cells\;mL^{-1}$ in summer, $4.1{\times}10^3\;cells\;mL^{-1}$ in autumn and $8.5{\times}10^3\;cells\;mL^{-1}$ in winter, respectively. In order to investigate factors influencing the total phytoplankton standing crops a multiple regression analysis was adopted for the correlation between standing crops and environmental factors. The coefficient of determination ($R^2$) value of the regression was 0.465, it showed that environmental factors which predominantly influenced phytoplankton standing crops were water temperature, COD, $NO_2-N$, $PO_4-N$, Discharge and pH. six stations could be divided into 3 groups based on similarity index in terms of environmental factors. In ANOVA analysis for physicochemical and biological factors, water temperature, chlorophyll a, silicate, phytoplankton standing crops were the same group differed little from stations. However, Station 1and 2 were grouped followed in dissolved oxygen, conductivity, COD, nitrite, nitrate, ammonia and phosphate, and Station 3, 4 and 5 were followed in dissolved oxygen, conductivity, pH and phosphate.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.14 no.2
• /
• pp.164-170
• /
• 1985

Yellowish modified wool, dithiocarbamate(DTC) wool, was synthesized by partial hydrolysis in 0.2 N-NaOH reacting with carbon disulfide to use as ${\alpha}-amylase$ immobilization matrix. ${\alpha}-amylase$ was immobilized reacting with sulfide group of DTC-wool by covalent binding within 1 hour. 0.5 gram of this preparation, $DTC-wool-{\alpha}-amylase$, contained 150 ug of enzyme protein and its specific activity was about 90% of the native one. General properties of $DTC-wool-{\alpha}-amylase$ were a little different from optimum temperature, optimum pH, heat stability, kinetic constants and activation energy. An apparent Michaelis constant and maximum velocity of $DTC-wool-{\alpha}-amylase$ were 5.56 mg/ml and 0.37 mg/ml. $min^{-1}$ respectively, while activation energy was 16.6 kcal/mole.