• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.195-202
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• 2023

Protease is a widely used enzyme particularly in the detergent industry. In this research, we aimed to isolate alkaline protease-producing bacteria for characterization as a laundry detergent additive. The screening of alkaline protease production was investigated on basal medium agar plus 1% skim milk at pH 11, with incubation at 30℃. The highest alkaline protease-producing bacterium was 6BS15-4 strain, identified as Bacillus gibsonii by 16S rRNA gene sequencing. While the optimum pH was 12.0, the strain was stable at pH range 7.0-12.0 when incubated at 45℃ for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60℃ and the enzyme was stable at 55℃ when incubated at pH 11.0 for 60 min. Metal ions K⁺, Mg²⁺, Cu²⁺, Na⁺, and Zn²⁺ exhibited a slightly stimulatory effect on enzyme activity. The enzyme retained over 80% of its activity in the presence of Ca²⁺, Ba²⁺, and Mn²⁺. Thiol reagent and ethylenediaminetetraacetic acid did not inhibit the enzyme activity, whereas phenylmethylsulfonyl fluoride significantly inhibited the protease activity. The alkaline protease from B. gibsonii 6BS15-4 demonstrated efficiency in blood stain removal and could therefore be used as a detergent additive, with potential for various other industrial applications.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.1
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• pp.33-38
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• 2004

This study was aimed at testing the gene expression of antioxidant enzymes and apoptosis genes for in vitro culture in porcine embryos produced by in vitro maturation/in vitro fertilization (IVM/IVF). Pocine preimplantation embryos obtainted from IVM/IVF can be successfully culture in vitro, but they are delayed or stop to develop at specific developmental stage. Many factors such as reactive oxygen species and apoptosis in an IVM/IVF system followed by in vitro culture influence the rate of production of viable blastocysts. Porcine embryos derived from IVM/IVF were cultured in the atmosphere of 5% $CO_2$ and 20% $O_2$ at $38.5^{\circ}C$ in NCSU23 medium. The patterns of gene expression for antioxidant enzymes and apoptosis genes during in vitro culture in pocine IVM/IVF embryos were examined by the modified semi-quantitative single cell reverse transcriptase-polymerase chain reaction (RT-PCR). Porcine embryos produced by in vitro procedures were expressed mRNAs for CuZn-SOD, GAPDH and GPX, whereas transcripts for Mn-SOD and catalase were not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell 16 cell and blastocyst, but p53 mRNA was not detected at any stages. The fas transcripts was only detected in blastocyst stage. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in vitro culture of porcine IVM/IVF embryos.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1546-1554
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• 2010

A bacterial strain capable of producing extracellular ${\alpha}$-galactosidase was isolated from a sample of sugarcane industrial waste. Microbiological, physiological, and biochemical studies revealed that the isolate belonged to Bacillus sp. Furthermore, based on a 16S rDNA sequence analysis, the new isolate was identified as Bacillus megaterium VHM1. The production of ${\alpha}$-galactosidase was optimized based on various physical culture conditions. Guar gum and yeast extract acted as the best carbon and nitrogen sources, respectively. The optimum pH was 7.5 and the enzyme remained stable over a pH range of 5-9. The enzyme was optimally active at $55^{\circ}C$ and thermostable with a half-life of 120 min, yet lost 90% of its residual activity within 120 min at $60^{\circ}C$. One mM concentrations of $Ag^2$, $Cu^2$, and $Hg^{2+}$ strongly inhibited the ${\alpha}$-galactosidase, whereas the metal ions $Fe^2$, $Mn^{2+}$, and $Mg^{2+}$ had no effect on the ${\alpha}$-galactosidase activity, and $Zn^{2+}$, $Ni^{2+}$, and $Ca^{2+}$ reduced the enzyme activity slightly. When treated with the B. megaterium VHM1 enzyme, the flatulence-causing sugars in soymilk were completely hydrolyzed within 1.5 h.

• Proceedings of the Korean Vacuum Society Conference
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• 2000.02a
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• pp.211-211
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• 2000

Carbon nanotubes (CNTs) have been spotlighted as one of promising field emission displays(FEDs). For the first time, to authors knowledge, we have developed the 9" color CNT-FEDs with the resolution of 240x576 lines. The 9" CNT-FEDs with diode-type and triode-type structures are presented. The well-dispersed CNT paste was squeezed onto the metal-patterned cathode glass. For the anode plate, the Y2O2S:Eu, ZnS:Ag,Cl low-voltage phosphors were printed for red, green, and blue colors, respectively. The vacuum-packaged panel maintained the vacuum level of 1x10-7 Torr. The uniform moving images vacuum-packaged panel maintained the vacuum level of 1x10-7 Torr. The uniform moving images were demonstrated at 2 V/um. High brightness of 800, 200, and 150cd/m2 was observed on the green, red, and blue phosphors at V/um, respectively. Field emission characteristics of a triode-type CNT-FED were simulated using a finite element method. the resultant field strength on the cathode was modulated by gate bias and emitted electrons were focused on the anode. A relatively uniform emission image was experimentally achieved at the 800V anode. A relatively uniform emission image was experimentally achieved at the 800V anode and the 50-180 V gate biases. Energy distribution of electrons emitted from CNTs was measured using an energy analyzer. The maximum peak of energy curve corresponded to the Fermi energy level of CNTs. The whole fabrication processed of CNT-FEDs were fully scalable and reproducible. Our CNT-FEDs has demonstrated the high potential of large-area and full-color applications with very low cost fabrication and low power consumption.

• Economic and Environmental Geology
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• v.2 no.4
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• pp.23-71
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• 1969

Sediment samples were taken at about half-mile intervals from all the inajor rivers draining the St. Austell granite mass. The minus 80 mesh(B.S.S.) fraction of each sample was analysed, using semiquantitative methods, for sodium, potassium, lithium, phosphorus, nickel, chromium, tin, tungsten, arsenic copper, zinc and lead. The work was carried out with the view to gaining further information as to the geographical distribution of such different granite facies as might axist, and to investigate the geochemical dispersion of these elements with relation to mineralisation in this area. The sesults confirm Exley's suggestion that the mass consists of two major granite intrusions, the earlier undifferentiated one is joined on the west by a later differentiated intrutive. During the work grid deviation maps proved particularly useful in obtaining data concerning the nature of the granite but frequency diagrams were not particularly helpful. All the known lode areas were associated with stream sediments containing anomalously high concentrations of lode metals and it is concluded that these high concentrations are due premarily to lode material transferred to the streams in the form of tailings lost during milling operations.

• Economic and Environmental Geology
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• v.28 no.3
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• pp.251-257
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• 1995

Three PEA (Pulverised Fuel Ash) samples, which were fresh, 17 and some 40 years weathered, were collected from two major British power plants. Batch leaching tests with these samples using distilled water and simulated industrial leachate showed higher amounts of element liberation from fresh ash, including Ca, Na, K, S (as $SO^{2-}_4$, $Cr_{total}$, Cu, Li Ni, Mo and CI and this seems to indicate their surface association and easier dissolution when contact with water. On the contrary Mg, Al, Ba, Si, V, As and Se do not show such readily leachable concentrations and these elements might be more associated with glass fraction in PFA particle rather than surface. Although element concentrations in the weathered ash are much lower than those in the initial leachate from the fresh ash, elements are still detected as resonable concentrations, with rather constant levels and this seems to demonstrate the element release from unstable glass phase of PFA particle. Fe, Ca, $Cr_{total}$, Cu, Ni, Zn and Hg were removed from the synthetic leachate by PFA and this is also confirmed by gain in solid PFA. The order of element retention is Meaford weathered ash > Drax weathered ash > Drax fresh ash in decreasing order and this conforms with the degree of weathering. Namely, the more wethered, the more wethered, the more effective in metal retention from the synthetic leachate.

• Journal of environmental and Sanitary engineering
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• v.15 no.4
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• pp.59-77
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• 2000

This study was carried out to investigate on several factors, which contaminative the water quality through the water pipe during feeding water, in 42 largescaled apart-ments(total 84 cases) and assayed the Volatile Organic Compounds(VOCs) and concen-tration of heavy metals that inflow and outflow in reservior water in Jeonnam area(Mokpo, Suncheon, Yeosu) from January 1999 to December 1999. The results obtained were summarized as follows ; 1. The quality of the water pipe composition in the order of frequency in the quality of water pipes were Copper(45.2%)> Zinc(38.9%)> Stainless steel(9.5%)> PVC(4.8%)> PM(2.4%) in observing 42 sites. All of the drain pipes were used the cast iron quality. 2. The result of certification curve from 12 items(17kind) of VOCs was $1.0-4.0{\mu{g}}/{\ell}$ range, a coefficient of correlation was shown 0.99 over. A MDL of each substance range was within $0.1-1.0{\mu{g}}/{\ell}$. 3. The result of the assay, 5 kinds(Viny chloride, Dichloromethane, Ethylbenzene, M,P-xylene, Styrene) of the VOCs of 14 kinds was not detected and the other items were detected slightly. The detection rate of one item and over in total VOCs samples, were 25.9% in inflow and 27.9% in outflow. And frequency of detect in inflow/outflow of THM(Chloroform, Bromodichloro-methane, Dibromochloromethane, Bromoform) were shown higher than 94.1%, 97.0% each stages. It comes to the conclusion that all of the samples were reason able for drinking water standards. 4. The coefficient of correlation were reasonable, it shown 0.999 over in $0.1-1.0{\mu{g}}/{\ell}$ of a measuring range conditions of 4kinds in organic substance(Zn, Cu, Fe, Mn). 5. The results were showed suitability in 78 cases(92.9%) and unsuitability in 6 cases (7.1%), in 84 cases of in organic substances. Compare to inflow stage, mean concentrations of heavy metal, were increased slightly in Zn, Cu, Fe except Mn than outflow stage. The result of the mean concentration in organic substance inflow and outflow in the apartment water tank using Pair-compared T-test, in 95% reliance index, were $0.179mg/{\ell}(0.151-0.307mg/{\ell})$ in Zinc, $0.136mg/{\ell}(0.113-0.230mg/{\ell})$ in Copper, $0.052mg/{\ell}(0.048-0.098mg/{\ell})$ in Fe, and there was a bit growing tendency but there was no differece in Mn. 6. The mean concentration of Copper which used Cu pipe as a water supply pipe in apartment were $0.216mg/{\ell}(0.161-0.338mg/{\ell})$ in case of the Zine pipe were $0.286mg/{\ell}(0.204-0.435mg/{\ell})$. It shows that the detection rate was more higher than the other material used in Cu or Zn as the water supply pipe. We supposed to Cu and Zn substance were gushing out water supply pipe.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.1
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• pp.35-41
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• 2006

Antibacterial activities of the trace elements in combination with the food additives were measured against 6 kinds of food-borne microorganisms such as Escherichia coli, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis and Pseudomonas fluorescens. The difference of antibacterial activity was not shown among the kinds of food additives, such as dextrin, gelatin and collagen. Zn and Ge in combination with food additives had strong antibacterial effect. Especially, $1\%$ zinc acetate in combination with $1\%$ gelatin was more effective against P. fluorescens and S. aureus than against Bacillus sp., E. coli and V. parahaemolyticus. Minimum inhibitory concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on S. aureus and P. fluorescens, and 1.0 mg/mL on E. coli, V. parahaemolyticus, B. cereus and B. subtilis. Minimum bactericidal concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on P. fluorescens and 1.0 mg/mL on E. coli, V. parahaemolyticus, S. aureus, B. cereus and B. subtilis. The zinc acetate in combination with gelatin showed stronger inhibitory effect in acidic range below pH 6.0, and remained active even after heat treatment at $121^{\circ}C$ for 15 min. In comparison with control, the viable cell counts of fish pastes, which were coated with the solution containing both $1\%$ zinc acetate and $3\%$ gelatin, were decreased by more than 100-fold until the storage of 7 days at $10^{\circ}C$. The results indicate that the combined use of zinc acetate and some food additives could prolong the shelf life of fish pastes by 8 days or more at $10^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.20 no.5
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• pp.519-526
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• 1992

The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

• Food Science of Animal Resources
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• v.22 no.3
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• pp.259-266
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• 2002

The egg shell membrane degrading isolated from soil was identified as Bacillus licheniformis by 16S rDNA identification method. A keratinase was isolated from the Baciilu licheniformis culture. DEAE-cellulose ion-exchange and Sephadex C-75 gel chromatograhies were used to purify the enzyme. The specific activity was increased 17.3-fold by the purification procedures. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and Sephadex G-75 chromatography indicated that the purified keratinase was monomeric and had a molecular weight of 65 kDa. The enzyme showed optimum activity at pH 9.0, and was stable above pH 9.0. The optimum temperature was 50$\^{C}$ and the enzyme was stable in the temperature ranges from 20$\^{C}$ to 50t. By the addition of 1 mM and 10 mM FeSO4, the activities of the enzyme were increased to 111$\pm$4.6% and 133$\pm$3.79%, respectively. The keratinase was an alkaline serine pretense because it was inhibited only by phenylmethylsulfonylfluorice (PMSF).