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Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents (세탁세제 첨가용 효소 개발을 위한 남극 해양세균 유래 저온성 단백질분해효소의 특성 연구)

  • Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.60-68
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    • 2018
  • A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

Characterization of a Novel Fibrinolytic Enzyme Produced from Bacillus subtilis BK-17 (Bacillus subtilis BK-17 유래 혈전용해 효소의 특성)

  • Hyun Bek;Lim Hak-Seob;Chung Kyung Kae;Choi Yung Hyun;Choi Byung Tae;Seo Min-Jeong;Kim Ji-Eun;Ryu Eun-Ju;Huh Man Kyu;Joo Woo Hong;Jeong Young Kee
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.987-993
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    • 2005
  • A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.

Hydrogeochemical Characterization of Groundwater in Jeju Island using Principal Component Analysis and Geostatistics (주성분분석과 지구통계법을 이용한 제주도 지하수의 수리지화학 특성 연구)

  • Ko Kyung-Seok;Kim Yongie;Koh Dong-Chan;Lee Kwang-Sik;Lee Seung-Gu;Kang Cheol-Hee;Seong Hyun-Jeong;Park Won-Bae
    • Economic and Environmental Geology
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    • v.38 no.4 s.173
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    • pp.435-450
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    • 2005
  • The purpose of the study is to analyze the hydrogeochemical characteristics by multivariate statistical method, to interpret the hydrogeochemical processes for the new variables calculated from principal components analysis (PCA), and to infer the groundwater flow and circulation mechanism by applying the geostatistical methods for each element and principal component. Chloride and nitrate are the most influencing components for groundwater quality, and the contents of $NO_3$ increased by the input of agricultural activities show the largest variation. The results of PCA, a multivariate statistical method, show that the first three principal components explain $73.9\%$ of the total variance. PC1 indicates the increase of dissolved ions, PC2 is related with the dissolution of carbonate minerals and nitrate contamination, and PC3 shows the effect of cation exchange process and silicate mineral dissolution. From the results of experimental semivariogram, the components of groundwater are divided into two groups: one group includes electrical conductivity (EC), Cl, Na, and $NO_3$, and the other includes $HCO_3,\;SiO_2,$ Ca, and Sr. The results for spatial distribution of groundwater components showed that EC, Cl, and Na increased with approaching the coastal line and nitrate has close relationship with the presence of agricultural land. These components are also correlated with the topographic features reflecting the groundwater recharge effect. The kriging analysis by using principal components shows that PC 1 has the different spatial distribution of Cl, Na, and EC, possibly due to the influence of pH, Ca, Sr, and $HCO_3$ for PC1. It was considered that the linear anomaly zone of PC2 in western area was caused by the dissolution of carbonate mineral. Consequently, the application of multivariate and geostatistical methods for groundwater in the study area is very useful for determining the quantitative analysis of water quality data and the characteristics of spatial distribution.

Separation of Ni and Fe from $H_2SO_4$ leaching solution of scrapped Fe-Ni alloy (Fe-Ni 합금(合金) 스크랩의 황산(黃酸) 침출액(浸出液)으로부터 Ni와 Fe의 분리(分離))

  • Yoo, Kyoung-Keun;Jha, Manis Kumar;Kim, Min-Seuk;Yoo, Jae-Min;Jeong, Jin-Ki;Lee, Jae-Chun
    • Resources Recycling
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    • v.17 no.1
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    • pp.80-87
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    • 2008
  • Cementation and solvent extraction processes were studied to separate nickel and iron ions from the $H_2SO_4$ leaching solution with 47 g/L $Fe(Fe^{2+}/Fe^{3+}=1.03),$, 23.5 g/L Ni and 0.90M $H_2SO_4$ which leached from Fe-Ni alloy. Iron powder was used as a reducing agent for the cementation of Ni ion from the leaching solution. The reduction percentage of Ni ion was $17{\sim}20%$ by adding 4 times stoichiometric amount of iron powder at $60{\sim}80$. This may result from the fact that the cementation of Ni ion occurred after the reduction of $Fe^{3+}$ to $Fe^{2+}$ and the neutralization of $H_2SO_4$ with iron powder. The cementation process was proved to be unfeasible for the separation/recovery of Ni ion from the leaching solution including $Fe^{3+}$ as a major component. $Fe^{2+}$ present in the leaching solution was converted to $Fe^{3+}$ for solvent extraction of Fe ion using D2EHPA in kerosene as a extractant. The oxidation of $Fe^{2+}$ to $Fe^{3+}$ was completed by the addition of 1.2 times stoichiometric amount of 35% $H_2SO_4$. 99.6% $Fe^{3+}$ was extracted from the leaching solution (23.5 g/L $Fe^{3+}$) by 4 stages cross-current extraction using 20 vol.% D2EHPA in kerosene. $NiSO_4$ solution with 98.5% purity was recovered from the $H_2SO_4$ leaching solution of scrapped Fe-Ni alloy.

Flavor Constituents in Enzyme Hydrolysates from Shore Swimming Crab and Spotted Shrimp (민꽃게 및 꽃새우 효소가수분해물의 풍미발현성분)

  • ;;Chi-Tang Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.787-795
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    • 2001
  • For the developing natural fisheries flavoring substances using crustacea, the flavor constituents of enzyme hydrolysates from shore swimming crab (crab) and spotted shrimp (shrimp) were investigated. In taste-active compounds of both enzyme hydrolysates, total free amino acid contents of crab and shrimp enzyme hydrolysates were 5,226.7 mg% and 8,757.3 mg%, respectively. The major amino acids were taurine, glutamic acid, proline, asparagine, glycine, alanine, valine, leucine, lysine anserine and arginine. As for ATP related compounds, AMP was the principal component and small amounts of IMP was detected in both enzyme hydrolysates. In the quarternary ammonium bases, betaine was the principal component (593.8mg%), and contents of TMAO and betaine in both samples were 60.7 mg% and 850.0 mg%, 124.1 mg% and 755.9 mg%, respectively. The major components were Na, K, P and Cl in inorganic ions. The major fatty acids of both sample were 14 : 0, 16 : 0, 16 : 1n7, 18 : 1n9, 20 : 5n3 and 22 : 6n3, and composition ration of n3 polyunsaturated fatty acids of were 27.8% and 28.5%, respectively. Total 99~109 volatile compounds were detected as a cooked odor of crab and shrimp enzyme hydrolysates by SDE apparatus/gas chromatography/mass spectrometry. The volatile flavor compounds identified from cooked crab enzyme hydrolysate were composed of 6 acids, 10 alcohols, 7 aldehydes, 11 ketones, 1 ester, 5 phenols, 4 benzenes, 22 hydrocarbons, 1 furan, 21 nitrogen containing compounds and 11 micellaneous compounds. And the volatile flavor compounds indentified from cooked shrimp enzyme hydrolysate were composed of 13 acids, 10 alcohols, 6 aldehydes, 10 ketones, 3 esters, 2 phenols, 5 benzenes, 36 hydrocarbons, 1 furan, 14 nitrogen containing compounds and 8 micellaneous compounds.

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Food Components Characteristic of Oysters Produced in Korea (국내 산지별 굴의 성분 특성)

  • Choi, Jong-Duck;Hwang, Seok-Min;Kang, Jin-Young;Kim, Sang-Hyun;Kim, Jeong-Gyun;Oh, Kwang-Soo
    • Journal of agriculture & life science
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    • v.46 no.6
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    • pp.105-115
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    • 2012
  • The detailed proximate, fatty/amino acid, mineral composition, texture, color, chemical and taste compounds of six oysters (four kinds of cultured oysters and two kinds of wild oysters) in Korea were investigated. Length and weight of the shell removed cultured and wild oysters were 4.7~5.1 and 3.0~4.2 cm, and 5.9~9.1 and 2.6~5.5 g, respectively. The proximate compositions were not significantly different between cultured and wild oysters. Amino nitrogen and volatile basic nitrogen content of these ones were 232.8~258.2 and 160.5~213.9 mg/100 g, 9.5~12.0 and 7.8~9.5 mg/100 g, respectively. As a texture characteristic of muscle, shearing force were 95~114 and 105~132 g. Amounts of total amino acids of cultured and wild oysters were 9,004~10,198 and 8,165~8,942 mg/100 g, respectively. Major amino acids and inorganic ions were aspartic acid, glutamic acid, proline, alanine, leucine, phenylalanine, lysine, arginine and K, Na, Ca, Fe, S, P, Zn. Major fatty acids of these ones were 16:0, 18:0, 16:1n-9, 18:1n-9, 22:1n-9, 16:4n-3, 20:5n-3 and 22:6n-3, and there was little difference between the two groups. Amounts of free amino acids of cultured and wild oysters extracts were 1,444~1,620 and 1,017~1,277 mg/100 g, respectively, and major ones were taurine, glutamic acid, glycine, alanine, tryptophan, ornithine and lysine. There is a little difference in glycine, tryptophan, ornithine and arginine contents, but TMAO and TMA contents were low in amount, and were not significantly different between the two groups.

Effects of Air Pollution on the Decline of Pinus thunbergii forest in Urban Industrial Area (도시공단의 대기오염이 해송의 쇠퇴에 미치는 영향)

  • Lee, Sung-Sik;Kim, Jeong-Woon;Lee, Chong-Kyu
    • Journal of agriculture & life science
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    • v.46 no.3
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    • pp.1-10
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    • 2012
  • This study was carried out to provide basal information on factors effected the decline of Pinus thunbergii by analysisof, the content of chlorophyll, and acid deposition through stepwise regression analysis. pH in throughfall at industrial area was 4.65 in the spring and 4.72 in the fall. On the contrary, pH at rural area was measured 5.32 in the spring and 5.34 in the fall. EC of rain fall at industrial area was $262.30{\mu}s$, However, there was $47.72{\mu}s$ at control area. Of anions, the concentration of ${NO_3}^-$ was $52.13mg/{\ell}$ at industrial area and $37.85mg/{\ell}$ at area. The concentration of ${SO_4}^{2-}$ was $57.89mg/{\ell}$ at industrial area and $36.21mg/{\ell}$ at area. Of the concentration of leaf chlorophyll in Pinus thunbergii, chlorophyll a was 0.2378 but control area was 0.4378. Also the content of chlorphyll b was 0.2097 and control area was 0.2345. The degree of decline of Pinus thunbergii forest was 2.97 at industrial area and 1.20 at area. We carried out a correlation analysis between the degree of decline and the concentration of ions, $SO_2$, and $NO_2$ in Pinus thunbergii forest. As a result, there was a negative correlation (r=-0.8672) between rain acids, and a positive correlation between $SO_2$ concentrations (r=0.8924) and between $NO_2$ concentrations (r=0.8428) in air. The correlations among acid depositions at level of 1% were pH (r=-0.8672), ${NO_3}^-$(r=0.6996), ${SO_4}^{2-}$(r=0.8497), $SO_2$ (r=0.8924), and $NO_2$ (r=0.8428).

Analysis of Correlation between Particulate Matter in the Atmosphere and Rainwater Quality During Spring and Summer of 2020 (봄·여름철 대기 중 미세먼지와 빗물 수질 상관성 분석)

  • Park, Hyemin;Kim, Taeyong;Heo, Junyong;Yang, Minjune
    • Korean Journal of Remote Sensing
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    • v.37 no.6_2
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    • pp.1859-1867
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    • 2021
  • This study investigated seasonal characteristics of the particulate matter (PM) in the atmosphere and rainwater quality in Busan, South Korea, and evaluated the seasonal effect of PM10 concentration in the atmosphere on the rainwater quality using multivariate statistical analysis. The concentration of PM in the atmosphere and meteorological observations(daily precipitation amount and rainfall intensity) are obtained from automatic weather systems (AWS) by the Korea Meteorological Administration (KMA) from March 2020 to August 2020. Rainwater samples (n = 216, 13 rain events) were continuously collected from the beginning of the precipitation using the rainwater collecting device at Pukyong National University. The samples were analyzed for pH, EC (electrical conductivity), water-soluble cations(Na+, Mg2+, K+, Ca2+, and NH4+), and anions(Cl-, NO3-, and SO42-). The concentration of PM10 in the atmosphere was steadily measured before and after the precipitation with a custom-built PM sensor node. The measured data were analyzed using principal component analysis (PCA) and Pearson correlation analysis to identify relationships between the concentration of PM10 in the atmosphere and rainwater quality. In spring, the daily average concentration of PM10 (34.11 ㎍/m3) and PM2.5 (19.23 ㎍/m3) in the atmosphere were relatively high, while the value of daily precipitation amount and rainfall intensity were relatively low. In addition, the concentration of PM10 in the atmosphere showed a significant positive correlation with the concentration of water-soluble ions (r = 0.99) and EC (r = 0.95) and a negative correlation with the pH (r = -0.84) of rainwater samples. In summer, the daily average concentration of PM10 (27.79 ㎍/m3) and PM2.5 (17.41 ㎍/m3) in the atmosphere were relatively low, and the maximum rainfall intensity was 81.6 mm/h, recording a large amount of rain for a long time. The results indicated that there was no statistically significant correlation between the concentration of PM10 in the atmosphere and rainwater quality in summer.

Comparative Analysis of Heavy Metal Contamination, Mineral Composition and Spectral Characteristics of White, Reddish Brown and Mixed Precipitates Occurring at Osip Stream Drainage, Gangwondo, South Korea (강원도 오십천 수계에 분포하는 백색침전물, 적갈색침전물 및 혼합침전물의 중금속 오염, 광물조성 및 분광학적 특성의 비교분석)

  • Lim, Jeong Hwa;Yu, Jaehyung;Shin, Ji Hye;Koh, Sang-Mo
    • Economic and Environmental Geology
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    • v.52 no.1
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    • pp.13-28
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    • 2019
  • This study analyzed precipitation environment, heavy metal contamination, and mineral composition of white, reddish brown and mixed precipitates occurring at the Osip stream drainage, Gangwondo. Furthermore, spectral characteristics of the precipitates associated with heavy metal contamination and mineral composition was investigated based on spectroscopic analysis. The pH range of the precipitates was 4.43-6.91 for white precipitates, 7.74-7.94 for reddish brown precipitates, and 7.59-7.9 for the mixed precipitates, respectively. XRF analysis revealed that these precipitates were contaminated with Ni, Cu, Zn, and As. The white precipitates showed high Al concentration compared to reddish brown precipitates as much as 3.3 times, and the reddish brown precipitates showed high Fe concentration compared to white precipitates as much as 15 times. XRD analysis identified that the mineral composition of the white participates was aluminocoquimbite, gibbsite, quartz, saponite, and illite, and that of reddish brown precipitates was aluminum isopropoxide, kaolinite, goethite, dolomite, pyrophyllite, magnetite, quartz, calcite, pyrope. The mineral composition of the mixed precipitates was quartz, albite, and calcite. The spectral characteristics of the precipitates was manifested by gibbsite, saponite, illite for white precipitates, goethite, kaolinite, pyrophyllite for reddish brown precipitates, and albite for the mixed precipitates, respectively. The spectral reflectance of the precipitates decreased with increase in heavy metal contamination, and absorption depth of the precipitates indicated that the heavy metal ions were adsorbed to saponite and illite for white precipitates, and goethite and magnetite for reddish brown precipitates.

Isolation of Wild Yeasts from Freshwaters and Soils in Nonsan Stream and Sapgyoho in Chungcheongnam-do, Korea, and Microbiological Characteristics of the Unrecorded Wild Yeasts (충남 논산천과 삽교호로부터 야생효모의 분리 및 국내 미기록 효모들의 균학적 특성)

  • Jang, Ji-Eun;Park, Seon-Jeong;Moon, Jeong-Su;Lee, Hyang Burm;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.49 no.3
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    • pp.337-349
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    • 2021
  • This study aimed to isolate wild yeasts from water and soil sample of the Nonsan stream and Sapgyoho (lake) in Chungcheonnam-do, Korea, and to further characterize previously unrecorded wild yeast strains. In total, 102 strains, representing 55 different species of wild yeast were isolated from 95 samples collected from the Jangseoncheon and Ipchoncheon of Nonsan stream in Jellabuk-do and Chungcheonnam-do. Among these, 33 strains were isolated from alkalophilic yeast extract-peptone-dextrose (YPD) medium (pH 9.0), and 9 strains were isolated concurrently on general YPD medium (pH 6.5) and alkalophilic medium. Seventeen strains of Cryptococcus laurentii were predominantly isolated. Additionally, 65 strains, representing 27 different species of wild yeast were isolated from 58 samples obtained from Sapgyoho (lake). Among the 82 isolated wild yeast strains, 8 strains, including Candida fructus JSC 72-1(JSL-GGU 015), had not previously been recorded. All 8 previously unrecorded yeasts were oval in shape except C. fructus JSC72-1(JSL-GGU-015), and only the Filobasidium chernovii JSC39-1(JSL-GGU-013) strain formed spores. All strains except Pseudosydowia eucalypti JSC23-6(JSL-GGU-012) grew well in yeast extract-peptone-dextrose (YPD) and yeast extract-malt extract media and grew in vitamin-free medium. Four strains, including P.eucalypti JSC23-6(JSL-GGU-012) grew well in 15% NaCl-containing YPD medium. F.chernovii JSC39-1(JSL-GGU-013) and Sirobasidium intermedium JSC7-3(JSL-GGU-014) assimilated lactose, and five strains, including F. chernovii JSC39-1(JSL-GGU-013) also assimilated starch. All strains were resistant to 800 ppm of Ca, Cu, Li, and Mg ions.