• Korean Journal of Soil Science and Fertilizer
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• v.42 no.1
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• pp.46-52
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• 2009

Deposition of atmospheric N that is depleted in $^{15}N$ has shown to decrease N isotope ratio ($^{15}N/^{14}N$,expressed as ${\delta}^{15}N$) of forest samples such as tree rings, foliage, and total soil-N. However, its effect on ${\delta}^{15}N$ of mineral soil-N which is biologically active N pool has never been tested. In this study, ${\delta}^{15}N$ of mineral N($NH{_4}^+$ and $NO_3{^-}$) in forest soils from organic and two depths of mineral soil layers (0 to 20 cm and 20 to 40cm depth) of Pinus densiflora stands located at two distinct areas (rural and industrial areas) in southern Korea was analyzed to investigate if there is any difference in ${\delta}^{15}N$ of mineral N between these areas. We also evaluated potential N loss of the study sites using ${\delta}^{15}N$ of mineral N. Across the soil layers, the ${\delta}^{15}N$ of $NH{_4}^+$ ranged from +8.9 to +24.8‰ in the rural area and from +4.4 to +13.8‰ in the industrial area. Soils from organic layer (+4.4‰) and mineral layer between 0 and 20 cm (+13.8‰) of industrial area showed significantly lower ${\delta}^{15}N$ of $NH{_4}^+$ than those of rural area (+8.9 and +24.3‰, respectively), probably indicating the greater contribution of $^{15}N$-depleted $NH{_4}^+$ from atmospheric deposition to forest in the industrial area than in the rural area. Meanwhile, ${\delta}^{15}N$ of $NO_3{^-}$ was not different between the rural and industrial areas, probably because ${\delta}^{15}N$ of $NO_3{^-}$ is more likely to be altered by the N loss that causes $^{15}N$ enrichment of the remaining soil N pool. Compared with the ${\delta}^{15}N$ of soil mineral N reported by other studies (from -10.9 to +15.6‰ for $NH{_4}^+$ and -14.8 to +5.6‰ for $NO_3{^-}$), the ${\delta}^{15}N$ observed in our study was substantially high, suggesting that the study sites are more subject to the N loss. It was concluded that $NH{_4}^+$ rather than $NO_3{^-}$ can conserve the ${\delta}^{15}N$ signature of atmospheric N deposition in forest ecosystems.

• Journal of Microbiology and Biotechnology
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• v.12 no.6
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• pp.972-978
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• 2002

Streptomyces sp. AD001 is a Gram-positive soil actinomycetes secreting an uncharacterized 2,4-dichlorophenol (DCP) oxidizing enzyme, whose activity is similar to the previously known Actinomycetes lignin-peroxidase (ALiP). This extracellular peroxidase was purified from Streptomyces sp. AD001 as a single protein band on an SDS-PACE by ammonium sulfate fractionation, Q-sepharose, concanavalin A, and Bio-Gel HTP column chromatographies. The molecular mass of the purified peroxidase was determined by SDS-PAGE to be 45.2 kDa, and 49.7 kDa with MALDI-TOF-MS, respectively. The highest level of peroxidase activity was observed at pH 7.5 and $30^{\circ}C$. The amino terminal sequence of the purified peroxidase (G-E-P-E-E-G-N-V-D-G-T-L) showed no significant homologies to my known proteins, suggesting that Streptomyces sp. AD001 may secrete a novel kind of bacterial peroxidase Initial rate kinetic data of the 2,4-DCP oxidation were best modeled with a random-binding bireactant system.

• Microbiology and Biotechnology Letters
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• v.10 no.3
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• pp.185-190
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• 1982

A strain of Streptomyces sp. (YS-40) which was able to produce penicillinase, was isolated from soil and the enzymatic characteristics of this enzyme were investigated. The crude enzyme was obtained with the fractionation by 80 % cold acetone. The optimal temperature and pH of this enzyme was 45$^{\circ}C$ and 5.0 respectively. The stable pH range of the enzyme was between pH 5.5 and 8.0 at 37$^{\circ}C$. By heat treatment at 6$0^{\circ}C$ and 8$0^{\circ}C$ for 10 min, the remained relative activities were about 50%, 30% respectively. The activity of the enzyme was inhibited by Cu$^{＋＋}$, $_Mn^{＋＋}$, Zn$^{＋＋}$ but Co$^{＋＋}$, Li$^{＋＋}$, $Ca^{＋＋}$, $Mg^{＋＋}$ $Ba^{＋＋}$ did not affect. Among 11 chemical reagents, ethylenedi aminetetra-acetic acid disodium salt (EDTA-2Na), sodium dodecyl sulfate (SDS) and sodium fluoride inhibited the enzyme activity.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.353-360
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• 1998

A bacterial strain PN33 producing large amounts of extracellular pectin lyase (PNL, EC 4.2.2.10) was isolated from soil. The isolated bacterium was identified as a strain of Bacillus sp. Production of PNL by the strain was induced only by pectins, with a higher degree of esterification, which had been added to the culture medium as a sole carbon source. The optimal medium for PNL production was determined to consist of 10 g pectin, 2 g yeast extract, 4 g $K_2HPO_4{\cdot}3H_2O$, 0.6 g $MgSO_4$, and 0.11 g $CaCl_2$ per liter (pH 7.0). The PNL activity in the culture supernatant reached the highest level of 132 mU/ml after 32 h cultivation at $37^{\circ}C$ in the optimal medium. The PNL produced was purified to homogeneity by ammonium sulfate fractionation (50~80%), and cation exchange and size exclusion chromatographies. The molecular mass of the enzyme was estimated to be approximately 52 kDa by SDS-PAGE. Almost the same mass was determined by nondenaturing PAGE, indicating that the functional enzyme had a monomeric structure. As expected, the PNL exhibited higher activities on the highly esterified pectins whereas it gave no detectable activity on polygalacturonic acid. The enzyme showed the highest activity at the acidic pH of 6.0, exceptional for a bacterial PNL. Maximum activity was measured at $40^{\circ}C$, although the stability f the purified enzyme was poor at this temperature. alcium (1 mM) was found to activate the PNL activity by $50\%$, and also remarkably increased the thermal stability f the enzyme. Phenylmethylsulfonylfluoride (PMSF) and iethylpyrocarbonate (DEPC) inhibited the PNL activity lmost completely at the concentration of 5 mM. This result ndicates that some serine and histidine residues of the nzyme may play an essential role for catalytic function of he enzyme.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.376-382
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• 1990

An extracellular cellulase producing bacterium YE-5 was isolated from soil, and identified as a Cellulomonas sp. by its taxonomical characteristics. The maximal activities of avicelase (0.35 units/ml), CMCase (3.18 units/ml), FPase (0.315 units/ml) and $\beta$-glucosidase (0.882 units/ml) were obtained when this strain was cultured for 48 hrs at $30^{\circ}C$ in a medium containing 0.8% (w/v) Solka floc, 0.06010 (wlv) urea, 0.1% (w/v) $K_2HP0_4$, 0.1% (w/v) $MgS0_4.7H2_0$, 0.2% (w/v) bacto peptone, 0.2% (w/v) yeast extract and pH 6.5. The cellulase was purified by ammonium sulfate fractionation, DEAE-Sepharose column chromatography and Sephadex 6-100 column chromatography from culture filtrate of Cellulomonus sp. YE-5. The molecular weights of purified avieelase, CMCase I, and CMCase II were estimated to be about 95,000 ~ 105,000, 46,000 ~ 47,000 and 120,000 ~ 125,000, respectively.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.259-264
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• 1988

Two forms of extracellular inulinase, designated as PI and PII were detected in the crude enzyme preparation from n species of Pseudomonas isolated from soil. PI and PII were purified to homogeneity by ammonium sulfate fractionation, DEAE Sephadex A-50 chromatography, Sephadex G-100 and Sephadex G-200 gel filteration. Both isoenzymes catalyzed specifically and endowise the cleavage of the $\beta$-2,1-fructofranoside linkage of inulin, and displayed no action upon sucrose, raffinose and levan. The optimal pH values for the PI and PII enzyme were pH 5.5 and 6.0, respectively and the highest activity of the two enzymes was observed at 55$^{\circ}C$. The Km values of PI and PII were calculated to be 2$\times$10$^{-3}$M and 5$\times$10$^{-3}$M, respectively.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.150-155
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• 1992

- Some properties of an extracellular cytosine deaminase excreted from an isolate from soil samples were examined after 20~80%' ammonium sulfate fractionation. The enzyme catalyzed the conversion of cytosine and 5-fluorocytosine into uracil and 5-fluorouracil by substrate specificity, respectively. The optimum temperature and storage time on the stability of the enzvme preparation were below $50^{\circ}C$ keeping above 90% of the residual activity and near 4 days keeping above 80% of the residual one in Tris-HCI buffer. The maximum activity was also obtained at 8.0 in pH and 37'C in temperature. The pHs and temperatures for enzyme activity ranged from 8.0~8.5 and from 37~$45^{\circ}C$. respectively. The presence of $Ag^-,Hg^{2-}, Zn^{2-}, Cu^{2-}, Sn^{2-}, \; or\; Pb^{2-}$ in the reaction mixture resulted in the marked inhibition in enzyme activity, but 1 mM of $K^+, Fe^[3+}, Mg^{2+}, \; or \; Na^+$. slightly increased the activity. The enzyme preparation was heavily affected by most of inhibitors tested such as 1 mM of EIITA, NaCN and pentachlorophenol, and completely inactivated by p-CMB and TCA of 1 mM, or 10 mM.

• Korean Journal of Pharmacognosy
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• v.36 no.1 s.140
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• pp.56-59
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• 2005

Urease plays an important role in the urea metabolism and the effect of urease activity on human and environment is enormous. For instance, urease acts as a virulence factor of the urinary and gastrointestinal tracts infections in human and animal, being involved in kidney stone formation, catheter encrusatation, pyelonephritis, ammonia encephalopathy, hepatic coma, and urinary tract infections. Widespread urease activity in soil induces a plant damage due to ammonia toxicity and pH increase. Therefore, urease activity regulation through urease inhibitors would lead to an enhanced efficiency of urea nitrogen uptake in plants and to the improved therapeutic strategies for ureolytic bacterial infections. To search for new inhibitory compounds on urease activity from herbs, MeOH extracts of herbs were screened. Among of them, the MeOH extracts of Areca catechu exhibited an excellent inhibitory effect on urease activity. Two compounds were isolated from the ethyl acetate fraction by the activity guided fractionation. Their chemical structures were identified as (+)-catechin(compound I) and allantoin(compound II) by spectroscopic evidence, respectively. Compound I showed a stronger inhibitory effect on urease activity than compound II.

• Journal of Korean Society on Water Environment
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• v.31 no.2
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• pp.159-165
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• 2015

The organic matter sources of phytoplankton and related environmental factors influencing algal bloom in Paldang reservoir were studied using nitrogen and carbon isotope ratio(${\delta}^{15}N$, ${\delta}^{13}C$). Phytoplankton samples for stable isotope analysis were collected from four points in reservoir using a plankton net. Physicochemical water quality, algal taxa and hydrological data were collected from published monitoring material. Phytoplankton samples were analyzed by IRMS. CN ratio of each sample was very similar to that of phytoplankton from literature cited. ${\delta}^{15}N$ of each sample was decreased during July. Mixing and dilution of nitrogen sources due to increment of influx by concentrated rainfall were considered as the main reason for the decline of ${\delta}^{15}N$. Based on analyzed ${\delta}^{15}N$ value of each sample, nitrogen source of Bughan river sample was presumed to come from soil. The nitrogen sources of Namhan river and Kyeongan stream samples seemed to be sewage or animal waste. Low ${\delta}^{15}N$ value in August (2012) seemed to be influenced by isotope fractionation due to the blooming of nitrogen-fixation blue-green algae (Anabaena spp.). Variation in ${\delta}^{15}N$ values particularly by blue-green algal bloom was considered the important factor for estimating the organic matter sources of phytoplankton.

• Journal of Microbiology and Biotechnology
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• v.6 no.1
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• pp.1-6
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• 1996

A bacterial strain NS70 producing an alkaline protease was isolated from soil samples taken near a hot spring and identified as Bacillus licheniformis by its morphological and physiological properties and cellular fatty acid analysis. The isolated alkaline protease was purified by ammonium sulfate fractionation, DEAE-, CM-, and Phenyl-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 32, 000 Da by sodium dodecylsulfate polyacrylamide gel electrophoresis. Its optimal pH and temperature for proteolytic activity against Hammarsten casein were 12 and $65^{\circ}C$, respectively. The enzyme was stable at alkaline pH range from 6.0 to 12.0, and fairly stable up to $65^{\circ}C$. The enzyme was inhibited by phenylmethylsulfonyl fluoride but not by EDTA and N-ethylmaleimide indicating that the enzyme is serine protease. Enzyme activity was markedly inhibited by $Hg^{2+}$ and $Cu^{2+}$. Autolytic phenomena were observed on purified protease NS70 but autolysis was reduced by the addtion of $Ca^{2+}$ ion or bovine serum albumin.