• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.584-592
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• 2004

A thermostable $\beta$-glycosidase gene, tfi $\beta$-gly, was cloned from the genomic library of Thermus filiformis Wai33 A1. ifi $\beta$-gly consists of 1,296 bp nucleotide sequence and encodes a polypeptide of 431 amino acids. It shares a strong amino acid sequence similarity with the $\beta$-glycosidases from other Thermus spp. belonging to the glycosyl hydrolase family 1. In the present study, the enzyme was overexpressed in Escherichia coli BL21 (DE3) using the pET21b(+) vector system. The recombinant enzyme was purified to homogeneity by heat treatment and a $Ni^{2+}$-affinity chromatography. Polyacrylamide gel electrophoresis (PAGE) showed that the recombinant Tfi $\beta$-glycosidase was a monomeric form with molecular mass of 49 kDa. The temperature and pH range for optimal activity of the purified enzyme were 80- $90^{\circ}C$ and 5.0-6.0, respectively. Ninety-three percent of the enzyme activity was remained at $70^{\circ}C$ after 12 h, and its half-life at $80^{\circ}C$ was 6 h, indicating that Tfi $\beta$-glycosidase is highly thermostable. Based on its K_m$, or $K_{cat}K_m$, ratio, Tfi $\beta$-glycosidase appeared to have higher affinity for $\beta$-D-glucoside than for $\beta$-D-galactoside, however, $K_{cat} for \beta$-D-galactoside was much higher than that for $\beta$-D-glucoside. The activity for lactose hydrolysis was proportionally increased at $70^{\circ}C$ and pH 7.0 without substrate inhibition until reaching 250 mM lactose concentration. The specific activity of Tfi TEX>$\beta$-glycosidase on 138 mM lactose at $70{^\circ}C$ and pH 7.0 was 134.9 U/mg. Consequently, this newly cloned enzyme appears to have a valuable advantage of conducting biotechnological processes at elevated temperature during milk pasteurization in the production of low-lactose milk.

• Journal of the Mineralogical Society of Korea
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• v.6 no.2
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• pp.57-79
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• 1993

Chuncheon nephrite, which was formed by the polymetasomatic alteration of dolomitic marble, can be classified into pale green, green, dark green, and grey types on the basis of their occurrence, mineralogical and textural characteristics. The nephrites consist obiefly of fibrous or hairlike(length/width ratio>10) cryptocrystalline(crystal width < $2{\mu}m$) tremolite, and include less amounts of micro-crystalline diopside, calcite, clinochlore, and sphene as impurities. The oriented and rather curved crystal aggregate, of nephritic tremolite are densely interwoven, resulting in a massive-fibrous texture which may explain the characteristic toughness of nephritic jade. The characteristic greenish color of the nephrite may be preferably related to Fe rather than Cr and Ni. However, the variation of color and tint in the Chuncheon nephrite also depends on the mineralogical and textural differences such as crystallinity, texture, and impurities. The chemical composition of the nephritic tremolite is not stoichiometric and rather dispersed especially in the abundances of Al, Mg, and Ca. Al content and Mg/Ca ratio for the nephritic tremolite are slightly increased with deepening in greenish color of the nephrite. Fe content in the nephritic tremolite is generally very low, but comparatively richer in the dark green nephrite. In nephritic tremolite, wide-chain pyriboles are irregularly intervened between normal double chains, forming a chain-width disorder. Most nephritic tremolites in the Chuncheon nephrite show various type of chain-width defects such as triple chain(jimthompsonite), quintuple chain (chesterite), or sometimes quadruple chain in HRTEM observations. The degree of chain-width disorder in the nephritic tremolite tends to increase with deepening in greenish color. Triple chain is the most common type, and quadruple chain is rarely observed only in the grey nephrite. The presence of pyribole structure in the nephritic tremolite is closely related to the increase of Al content and Mg/Ca ratio, a rather dispersive chemical composition, a decrease of relative intensity in (001) XRD reflection, and an increase in b axis dimension of unit cell. In addition, the degree and variation of chain-width disorder with nephrite types may support that an increase of metastability was formed by a rapid diffusion of Mg-rich fluid during the nephrite formation.

• Korean Journal of Agricultural and Forest Meteorology
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• v.7 no.4
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• pp.265-273
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• 2005

Chemical properties and heavy metal concentrations of forest soils of four abandoned coal mine lands affected by coal mining activities in the Mungyeong area were investigated to provide basic information for revegetation of abandoned coal mine lands. Soil pH in abandoned coal mine lands ranged from 5.30 to 6.76 it in the control site was 5.23. Contents of organic matter and total N in abandoned coal mine lands were $4.46\~7.19\%\;and\;0.07\~0.15\%$, respectively. Available P contents were 6.54 for A (Samchang), 6.52 for B (Bongmyeong),3.94 fur C (Kabjung), 5.45 mg/kg for D (Danbong coal mine land) and 5.25 mg/kg for the control site, which had a positive correlation with soil pH. Contents of exchangeable Ca, Mg, K and Na in abandoned coal mile lands averaged 196.1, 88.7, 88.2 and $10.2cmol^+/kg$, with a range of $132.1\~242.1,\;24.2\~138.\; 64.9\~120.8\;and\;8\~12.2cmol^+/kg$, respectively. Those of the control site were 192.8, 95.8, 104 and $21.2 cmol^+/kg$, respectively. Heavy metals such as Al, As, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn of forest soil in abandoned coal mine lands have a larger content than those of the control site. Al, Mn and fb content was especially high in abandoned coal mine lands. The Al content of forest soil in abandoned coal mine lands ranged from 397 to 917 ppm, which was considered to be high enough to inhibit tree growth. Therefore, it is suggested that soils of abandoned coal mine lands contaminated by mining activities need to be properly treated for remediation of environmental problems.

• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.239-255
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• 1997

Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.114-120
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• 2002

A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2＋$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.1
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• pp.80-85
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• 2004

The purpose of this experiment was to assess seasonal variation of feed utilization by feed sources and to obtain information on the use of feed resources by comparing seasonal changes of dry matter intake, digestibility, nitrogen balance and feeding behavior in spotted deer (Cervus nippon) fed forest by-product silage (FBS) and corn silage (CS). Dry matter intake (DMI) of FBS was higher than that of CS in both winter and summer. While DMI of both diets was higher in summer, this was not significant at the 5% level. In contrast to DMI, digestible dry matter intake (DDMI) was higher for CS than for FBS in both seasons, but the difference was not significant. Digestibility of dry matter and crude protein was significantly higher (p<0.01) for CS than for FBS, whereas digestibility of crude fiber was significantly higher (p<0.01) for FBS than for CS in both seasons. Seasonal digestibility of dry matter and crude fiber for FBS was significantly greater (p<0.01) in summer than in winter: In summer, seasonal digestibility was 57.2% for dry matter and 55.5% for crude fiber, and in winter, 50.8% for dry matter and 30.7% for crude fiber. On the other hand, seasonal digestibility of crude protein was higher (p<0.01) in winter (42.1%) than in summer (32.3%). No significant difference (p>0.05) was found between the two seasons and diets for nitrogen intake (NI), 18.7 g/d in summer and 19.4 g/d in winter for FBS, 17.7 g/d in summer and 17.7 g/d in winter for CS. Fecal nitrogen was higher (p<0.01) for FBS than for CS and varied little seasonally. There was significant difference (p<0.01) between two seasons in urinary nitrogen, which was little difference between two diets. Retained nitrogen (RN) was different significantly (p<0.01) between two diets in both seasons, but there was little difference between seasons. Deer usually spent longer time on eating FBS than eating CS. Eating FBS took 221 min in summer and 187 min in winter, whereas eating CS took 113 min in summer and 109 min in winter. Deer spent less time on eating food in winter than in summer. Time spent on rumination was longer for FBS than for CS: for FBS, 504 min in summer and 456 min in winter, for CS, 423 min in summer and 279 min in winter. Time varied seasonally with both diets.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.638-643
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• 2007

The full encoding sequence for human type II hexokinase (HXK II) was cloned into the E. coli expression vector pET 21b and expressed as a C-terminally hexahistidine-tagged protein in the BL2l (DE3) strain. The IPTG-induced HXK II approximately accounted for 17% of the total E. coli proteins, and 81% of HXK $II_{6{\times}His}$ existed in inclusion bodies. To improve the production of soluble recombinant HXK II protein, in the functionally active form, we used low temperature, and the osmotic stress expression method. When expressed at $18^{\circ}C$, about 83% of HXK $II_{6{\times}His}$ existed in the soluble fraction, which amounted to a 4.1-fold yield over that expressed at $37^{\circ}C$. The soluble form of HXK $II_{6{\times}His}$ was also highly produced in the presence of 1M sorbitol under the standard condition $(37^{\circ}C)$, which indicated that temperature downshift and low water potentials were required to improve the yield of active recombinant HXK II protein. The expressed protein was purified by metal chelate affinity chromatography performed in an IDA Excellose column charged with $Ni^{2+}$ ions, resulting in about 40mg recombinant HXK II protein obtained with purity over 89% from 51 of E. coli culture. The identity of HXK $II_{6{\times}His}$ was confirmed by Western blotting analysis. Taken together, using the stress-governed expression described in this study, human active HXK II can be purified in sufficient amounts for biochemical and biomedical studies.

• Journal of Korean Society for Atmospheric Environment
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• v.22 no.5
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• pp.590-603
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• 2006

Size-and time-resolved aerosol samples were collected using an eight-stage Davis rotating unit for monitoring (DRUM) sampler from 23 March to 29 April 2001 at Gosan, Jeju Island, Korea, which is one of the super sites of Asia-Pacific Regional Aerosol Characterization Experiment(ACE-Asia). These samples were analyzed using synchrotron X-ray fluorescence for 3-hr average concentrations of 19 elements including Al, Si, S, Cl, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Br, Rb, and Pb. The size-resolved data sets were then analyzed using the positive matrix factorization(PMF) technique to identify possible sources and estimate their contributions to particulate matter mass. PMF analysis uses the uncertainty of the measured data to provide an optimal weighting. Twelve sources were resolved in eight size ranges($0.09{\sim}12{\mu}m$) and included continental soil, local soil, sea salt, biomass/biofuel burning, coal combustion, oil combustion, municipal incineration, nonferrous metal source, ferrous metal source, gasoline vehicle, diesel vehicle, and volcanic emission. The PMF result of size-resolved source contributions showed that natural sources represented by local soil, sea salt, continental soil, and volcanic emission contributed about 79% to the predicted primary particulate matter(PM) mass in the coarse size range ($1.15{\sim}12{\mu}m$) while anthropogenic sources such as coal combustion and biomass/biofuel burning contributed about 58% in the fine size range($0.56{\sim}2.5{\mu}m$). The diesel vehicle source contributed mostly in ultra-fine size range($0.09{\sim}0.56{\mu}m$) and was responsible for about 56% of the primary PM mass.

• Journal of Plant Biology
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• v.1 no.1
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• pp.11-20
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• 1958

I. Assumption: Mo-kam-ju, Koelreuteria paniculata is a woody plant mainly cistributed over Northern China. Its sporadic growth along the beaches of Korea and Japan is assumed to have been sowed by the seeds transported adrift on the current across the ocean. II. Use and Growth of the Plant: In China, this plant has been cultivated from early times mainly at temples, it seeds being used as rosaries, its flowers for yellow dyes and medicine for the eyes, and its leaves for black dyes. In Korea and Japan, these plants have been cultivated at temples and used as the material for rosaries. No natural growth of these plants was reported until 1919. III. Discoveries: In this domestic area, Dr. Chung Tae Hyun discovered the plant on the beach between Cho-Do and Chang-san-kot, Hwang-Hae Prov. in 1920. The reporter discovered them on the beach at Buk-Ni, Duk-jok-Do in 1948, at An-Hung in 1956 and on the beach at An-min-Do in 1957. In the Japan area, it was discovered for the first time twenty years ago, mainly along the coast line of the Japan Sea and some along the eastern coast line, at Subo, Yamaguchi Prefecture facing the Pacific Ocean. IV. Study and Experiment: A. Seed The seed coat is thick, non-permeable and floatable. A number of seeds were immersed in artificial sea water and fresh water separately. The seeds remained there for a hundred and forty-five (145) days from April 6, 1957 to August 29, 57. Thirty one seeds out of the fifty immersed in salt water and twenty seeds out of the fifty immersed in fresh water remained on the surface of the water, proving them to be non-permeable to both sea and fresh water. Of course, these had retained their germinating capabilities. Five (5) seeds out of twenty from the fresh water and six (6) out of thirty-one from the sea water were successfully germinated after a hole had been drilled in the seed coat and they had been planted. Thus their floating capability, non-permeability and germinating capability after a possible 145 day trip on the flowing currents has been proved satisfactory according to the assumption made above. B. Current As shown in the Data 2 and 3, the sea current initiated in the Pohai Sea flows westward down along the coast line of Korea reaching the southern part during the autumn and winter seasons. This fact also is in favor of the reporter's assumption. V. Discussion and Conclusion: The reporter concludes that, as discussed above, Koelreuteria paniculata which originates in Northern China is transported adrift on the flowing sea current to our western coast line, and also the coast lines of Shantung and Kuangtung of China and it is germinated on the sandy beaches forming new plants. Thus, the seeds drifted down on the southern beach of Korea and have been transported to the Japanese coast, adrift on the Tae-Ma current. Upon fruition, the seeds of the plants which settle on the coasts of western Korea and Japan will migrate to new places. It can be, however, assumed that while the thickness and nonpermeability of the coat enables the long travel in the water, this also can compose a difficulty in germination, consequently in developing a new distribution of this species.

• Journal of Food Hygiene and Safety
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• v.26 no.3
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• pp.273-282
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• 2011

This study established hazards which may cause risk to human at farm during cultivation stage of paprika. Samples of plants (paprika, leaf, stem), cultivation environments (water, soil), personal hygiene (hand, glove, clothes), work utensils (carpet, basket, box) and airborne bacteria were collected from three paprika farms (A, B, C) located in Western Gyeongnam, Korea. The collected samples were assessed for biological (sanitary indications and major foodborne pathogens), chemical (heavy metals, pesticide residues) and physical hazards. In biological hazards, total bacteria and coliform were detected at the levels of 1.9~6.6 and 0.0~4.610g CFU/g, leaf, mL, hand or 100 $cm^2$, while Escherichia coli was not detected in all samples. In major pathogens, only Bacillus cereus were detected at levels of ${\leq}$ 1.5 log CFU/g, mL, hand or 100 $cm^2$, while Staphylococuus aureus, Listeria monocytogenes, E. coli O157 and Salmonella spp. were not detected in all samples. Heavy metal and pesticide residue as chemical hazards were detected at levels below the regulation limit, physical hazard factors, such as insects, pieces of metal and glasses, were also found in paprika farms. Proper management is needed to prevent biological hazards due to cross-contamination while physical and chemical hazards were appropriate GAP criteria.