In this study, we produced the recombinant lunasin peptide using E. coli and P. pastoris, and evaluated biological activity of the recombinant lunasin peptide. Lunasin peptide was produced from E. coli transfected with pPGEX-lunasin expression vector and P. pastoris GS115 transfected with pPIC-lunasin expression vector. These recombinant lunasin peptides were similar to the synthetic lunasin peptide in the identification by LC-ESI-MS. In addition, the recombinant lunasin peptide from E. coli and P. pastoris was bound in the chromatin, and inhibited histone acetylation and the activity of histone acetyltransferase. These findings suggest that the production of the lunasin peptide using E. coli and P. pastoris will be useful for industrial utilization of lunasin peptide.
In this study, we produced the recombinant lunasin peptide using E. coli and P. pastoris, and evaluated biological activity of the recombinant lunasin peptide. Lunasin peptide was produced from E. coli transfected with pPGEX-lunasin expression vector and P. pastoris GS115 transfected with pPIC-lunasin expression vector. These recombinant lunasin peptides were similar to the synthetic lunasin peptide in the identification by LC-ESI-MS. In addition, the recombinant lunasin peptide from E. coli and P. pastoris was bound in the chromatin, and inhibited histone acetylation and the activity of histone acetyltransferase. These findings suggest that the production of the lunasin peptide using E. coli and P. pastoris will be useful for industrial utilization of lunasin peptide.
In this study, bacterial growth was assessed by flow cytometry analysis of fluorescent probes-stained bacteria. Flow cytometry has many advantages of rapid analytical time, a low standard deviation, and highly sensitive detection of live and Dead E.coli over colony forming assay. When untreated bacteria were stained by using Thiazole Orange (TO) and Propidium Iodide (PI), double staining had a short analytical time as compared with that of single staining while its error rate was similar to that of single staining. Through double staining experiments, it was determined that optimal concentrations for TO and PI staining were 420 nM and 9.6 μM, respectively.
Limonoid는 항바이러스 및 항균제로써의 치료적인 목적으로 널리 연구되고 있는 성분으로 감귤에서 풍부하게 존재한다. 그러나 성분 자체의 쓴맛으로 인하여 기호성이 저하되므로 이를 해결하는 노력이 필요하며 제품 개발이 요구되고 있는 실정이다. 이러한 쓴맛을 제거할 수 있는 훌륭한 효소로써 LUGT가 주목되고 있으며, 이에 효소의 특성화 연구를 위하여 분리 및 정제를 시도하였다. Plasmid vector인 pET30a(+)을 사용하여 대장균에서
Sequential passes through columns were used to select phages that displays ligands for dextran (-1,6 linked linear chains) from a phage antibody library. Those phages that bound to the in each iteration were replicated in E. coli. A phage preparation isolated on the third round selection produced 5.4 nephelos turbidity units (NTU) in a dextran specific immunonephelometric assay, a 2.2 fold higher value than the phage preparation from the first round selection. This phage gave normalized intensity (N.I.) in a dip-stick assay against high molecular size dextran (T2000, 30\;{\pm}\;610^4\beta-galactosidase linked assay and polymerase chain reaction.
Using high voltage electric fields induced by high voltage AC (10-12 kV/cm, 20 kHz) and pulsed (20-30 kV/cm, 40 Hz) electric field generator as a semipermanent and environment-friendly disinfecting apparatus, the disinfection effect of coliform group in the effluent of sewage plant was investigated. The effects of electric field strength, treatment time, discharge area of a discharge tube, water quality factors (electric conductivity, pH and SS) on its death rate were examined. The death rate of coliform group was increased with increasing electric field strength and treatment time. For AC and pulsed electric field generator, the critical electric field strength was 6 kV/cm and 2 kV/cm, respectively, and the critical treatment time was 5 min and 2 min, respectively, regardless of electric field strength. Comparing the death rate of coliform group by AC and pulsed electric fields used in this study, its death rate was higher for the latter than the former, but did not increase linearly with increasing electric field strength. The results obtained for the effects of discharge area, electric conductivity, pH and SS on the death rate of coliform group using AC electric field (12 kV/cm, 20 kHz) were as follows: its death rate showed the trend to increase linearly with increasing discharge area; for the effect of electric conductivity, its death rate was increased with increasing electric conductivity, regardless of ionic species, increased with increasing cationic valency, but was similar between the same cationic valency; the pH 5~9 used in this study did not affect its death rate; its death rate was decreased with increasing SS concentration.
This study was carried out to investigate the effect of electrochemical (EC) disinfection of artificial wastewater contaminated by Escherichia coli culture. Circulated batch type electrochemical disinfection system using three plates electrodes was used. Also, the several factors (pH, ORP, DO, temperature, current, conductivity) were measured in order to investigate the fundamental design factor in the EC disinfection system. It was demonstrated that the EC process was highly effective for wastewater disinfection. At the constant voltage, the disinfection efficiency was increased according to time. The disinfection efficiency and current increased as the increase of voltage. The variation of conductivity was a little related to the variation of CFU (colony forming units). The differences in disinfection efficiency according to the ice pack and the variation of electrodes were not occurred. The EC disinfection efficiency and current increased according to the increase of circulating flow rate.
The experiment was carried out to determine the distribution of total coliform the translocation of heavy metals and the salt accumulation in soils following the supply of ground water, the domestic wastewater and the ultraviolet treatment of domestic wastewater during the spinach and the chinese cabbage cultivation. There were not much changes in the Total-N, Total-P and cations in soils following the ultraviolet treatment of the domestic wastewater. However, the density of total coliform was dramatically reduced from 894MPN/100mL to 5MPN/100mL. The diagnosis of composition of soil after the harvest of chinese cabbages and spinach has shown that the concentration of Na+ was 3-4 times higher in plot using domestic wastewater than in plot using ground water. When domestic wastewater and ground water were used in growing spinach md chinese cabbage at 50% each, the application of chemical fertilizers reduced about 25% to 50% compared to the criteria set down by the Rural Development Administration and there was not significant difference in terms of harvest. Using non-ultraviolet treatment of domestic wastewater directly on spinach and chinese cabbage has resulted in excessive density of total coliform at 25,000MPN/100mL. Even when ultraviolet treatment, the density was still high at 2,000MPN/100mL. The high density of total coliform even following ultraviolet treatment of domestic wastewater is considered to be caused by photo-reactivation of micro-organism. When reusing domestic wastewater, the application of sterilization such as ultraviolet, ultrasonic and electron-beam for public health and hygiene reasons may provide safe supply of agricultural water.