• Korean Journal of Agricultural Science
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• v.14 no.1
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• pp.144-155
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• 1987

A potent fungus producing milk clotting enzyme with fairly weak proteolytic activity was isolated from various soil and sewage, which the selected strain, SA-101, was identified as Mucor sp. with microbiological characteristics. Its milk clotting enzyme production was maximized when grown on 10g of wheat bran media added to 8ml of tap water containing 0.1M HCl for 60hrs at $30^{\circ}C$. This enzyme production was stimulated by addition of 6% lactose, 0.05% NaCl and reached a maximal level of 9810 unit/g wheat bran. The crude enzyme product could be produced effectively by salting out with ammonium sulfate fractionation and lyophilization. The ratio of milk clotting activity to proteolytic activity of crude enzyme product was lower than Hansen rennet, but resembled to Meito rennet. The optimal temperature of milk clotting activity of crude enzyme product was abound $60^{\circ}C$ on a substrate of 10% reconstituted skim milk containing 1/100M $CaCl_2$.

• Journal of Soil and Groundwater Environment
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• v.23 no.5
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• pp.26-36
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• 2018

Total petroleum hydrocarbon (TPH) is a mixture of various oil substances composed of alkane, alkene, cycloalkane, and aromatic hydrocarbons (benzene, toluene, ethylbenzene, xylene, etc.). In this study, we investigated 92 groundwater wells around 36 gas stations to evaluate distribution characteristics of petroleum hydrocarbons. Groundwater in the wells was sampled and monitored twice a year. The fraction analysis method of TPH was developed based on TNRCC 1006. The test results indicated aliphatic and aromatic fractions accounted for 28.6 and 73.8%, respectively. The detection frequencies of TPH in the monitoring wells ranged in 21.6 - 24.2%. The average concentration of TPH was 0.11 mg/L with the concentration range of 0.25~0.99 mg/L. In the result of TPH fraction analysis, in aliphatic fractions were 19% (C6-C8 : 0.2%, C8-C10 : 0.4%, C10-C12 : 0.4%, C12-C16 : 0.5%, C16-C22 : 1.0%, C22-C36 : 16.6%), and aromatic fractions were 81% (C6-C8 : 1.1%, C8-C10 : 0%, C10-C12 : 2.9%, C12-C16 : 0.3%, C16-C22 : 4%, C22-C36 : 66.8%). Fractions of C22-C36 were detected in about 83% of the monitoring wells, suggesting non-degradable characteristics of hydrocarbons with high carbon content.

• Journal of Soil and Groundwater Environment
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• v.24 no.3
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• pp.24-35
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• 2019

It is known that ${\delta}^{13}C_{DIC}$ (carbon-13 isotope of dissolved inorganic carbonate (DIC) ions) of water increases when dissolved $CO_2$ degases. However, ${\delta}^{13}C_{DIC}$ could decrease when the pH of water is lower than 5.5 at the early stage of degassing. Laboratory experiments were performed to observe the changes of ${\delta}^{13}C_{DIC}$ as $CO_2$ degassed from three different artificial $CO_2$-rich waters (ACWs) in which the initial pH was 4.9, 5.4, and 6.4, respectively. The pH, alkalinity and ${\delta}^{13}C_{DIC}$ were measured until 240 hours after degassing began and those data were compared with kinetic isotope fractionation calculations. Furthermore, same experiment was conducted with the natural $CO_2$-rich water (pH 4.9) from Daepyeong, Sejong City. As a result of experiments, we could observe the decrease of DIC and increase of pH as the degassing progressed. ACW with an initial pH of 6.4, ${\delta}^{13}C_{DIC}$ kept increasing but, in cases where the initial pH was lower than 5.5, ${\delta}^{13}C_{DIC}$ decreased until 6 hours. After 6 hours ${\delta}^{13}C_{DIC}$ increased within all cases because the $CO_2$ degassing caused pH increase and subsequently the ratio of $HCO_3{^-}$ in solution. In the early stage of $CO_2$ degassing, the laboratory measurements were well matched with the calculations, but after about 48 hours, the experiment results were deviated from the calculations, probably due to the equilibrium interaction with the atmosphere and precipitation of carbonates. The result of this study may be not applicable to all natural environments because the pressure and $CO_2$ concentration in headspace of reaction vessels was not maintained constant as well as the temperature. Nevertheless, this study provides fundamental knowledge on the ${\delta}^{13}C_{DIC}$ evolution during $CO_2$ degassing, and therefore it can be utilized in the studies about carbonated water with low pH and the monitoring of geologic carbon sequestration.

• Korean Journal of Environmental Agriculture
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• v.19 no.4
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• pp.319-323
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• 2000

This study was conducted to compare fractionations and availability of heavy metal in paddy soils near five abandoned mining areas. The sequential extraction procedure was used to fractionate the heavy metals in soils into the designated from water $soluble(H_2O)$, $exchangeable(0.5M\;KNO_3)$, organically bound(0.5M NaOH), $oxide/carbonate(0.05M\;Na_2-EDTA)$, and $sulfide/residual(4M\;HNO_3)$. EDTA and $HNO_3$ extractable of Cd, Pb, and Zn, and NaOH and $HNO_3$, extractable of Cu were predominant chemical forms. The ratio of $H_2O+KNO_3$ extractable of Cd, Zn, Cu, and Pb were 25.1, 8.7, 4.0, and 0.4%, respectively. The ratio of $H_2O+KNO_3$ extractable heavy metal were negatively correlated with soil pH, while $EDTA+HNO_3$ extractable heavy metal were positively correlated. The most consistent distribution patterns were found when the soil samples were grouped according to their total contents. Specially, the ratio of $H_2O+KNO_3$ extractable heavy metal were higher as total contents of heavy metal were increased. The ratio of $H_2O+KNO_3$ extractable heavy metal(Cd 1.06, Cu 0.15, Pb 0.01, and Zn 0.05%) were lower at the high soil pH than those(Cd 31.31, Cu 4.06, Pb 1.75, and Zn 10.16%) at the low level. Compared to other chemical forms, the degree of contribution for $KNO_3$ extractable form to the Cd uptake to brown rice was high, whereas that for EDTA and $HNO_3$ extractable forms were high to the Zn.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.4
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• pp.284-291
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• 2001

To study whether N isotope composition (${\delta}^{15}N$) of crop reflects the kind of fertilizer (chemical or organic) applied to field, a pot experiment was conducted. Corn (Zea mays L.) was cultivated under greenhouse conditions for 70 days. Composted pig manure and urea were applied at 0 and 0 (C0U0), at 0 and 300 (COU2), at 300 and 0 (C2U0) and at 150 and $150kg\;N\;ha^{-1}$ (C1U1), respectively. The ${\delta}^{15}N$ values of composted pig manure and urea were + 13.9‰ and -2.3‰, respectively. The ${\delta}^{15}N$ values of whole parts (roots + stems + leaves + grains) were + 12.7, + 12.9, + 14.0 and + 13.0‰ for C0U0, C0U2, C2U0 and C1U1 treatments, and were not significantly affected by the application of isotopically different N sources (P<0.05). However, leaves or grains showed significantly (P<0.05) different ${\delta}^{15}N$ values between treatments. The ${\delta}^{15}N$ values of leaves and grains were + 14.3 and + 16.2‰ for C2U0, +13.2 and +13.9‰ for C0U0, +10.1 and + 12.6‰ for C1U1 and +10.1 and +12.4‰ for C0U2 treatments. The different ${\delta}^{15}N$ values of corn from the values of N sources (compost and urea) applied to soil showed that the ${\delta}^{15}N$ values of corn were affected not only by the isotope composition of N source, but also by N pool mixing and isotope fractionation accompanying N transformation. This study suggests that although the ${\delta}^{15}N$ values of crop are not identical to the ${\delta}^{15}N$ values of N sources applied to fields, the application of isotopically different N sources such as compost and chemical fertilizer may result in qualitative difference in ${\delta}^{15}N$ values of crop.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.262-267
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• 2007

To access the natural product antibiotics produced by uncultured microorganisms, six cosmid libraries of DNA extracted directly from soil samples (environmental DNA, eDNA) were constructed and screened for the production of antibacterial active molecules. Of the approximately 60,000 clones screened, one antibacterial clone (YS92B) was detected. Ethyl acetate extracts of clone YS92B showed antibacterial activity against various pathogenic bacteria (Listeria monocytogenes, Bacillus subtilis, Pseudomonas syringae, Xanthomonas campestris pv. oryzae, Staphylococcus epidemis). Active constituents from cultures of YS92B were isolated and purified using a bioassay-guided fractionation against B. subtilis through a series of procedures (ethyl acetate extraction, Sephadex LH20 column chromatography, High Performance Liquid Chromatography). NMR (Nuclear Magnetic Resonance) spectral analysis of a major antibacterial active YS92B-VII indicated that it is a lauric acid linked to tyrosine. This report describes the characterization of antibacterially active long chain N-acyl derivatives of tyrosine that are produced by eDNA clones hosted in Escherichia coli from Korean soils.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.208-214
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• 1994

We screened many species from a wide variety of bacterial genera for a new type II restriction endonuclease. The purification and characterization of SolI from a soil isolate, Streptoverticillium olivoverticillatum are described here. The enzyme turned out to be an isoschizomer of BamHI. It recognized the hexanucleotide sequence of 5'-G$\downarrow$GATCC-3' and cleaved as in dicated by the arrow, generating a 4 base 5' extension. Unlike its isoschizomer, BamHI, the activity was sensitive to dam methylation within the recognition sequence. Following ammonium sulfate fractionation of the crude extract, heparin-agarose and Affi-gel Blue column chromatography were employed to purify the enzyme. SolI required at least 0.2 mM of $MgCl_2$ for the cleavage to occur. The enzyme exhibited its maximal activity in the absence of NaCl, but was inhibited completely in the presence of 120 mM NaCl. The pH and temperature optima for activity were pH 8.6 and $40^{\circ}C$, respectively. The molecular weight of SolI was estimated to be 43,000 Da by Superose-12 gel filtraion chromatography.

• Microbiology and Biotechnology Letters
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• v.5 no.4
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• pp.177-184
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• 1977

Arylsulfatase catalyzes the release of SO$\sub$4//sup2/- from sulfate esters of simple phenols. Arylsolfatase occurs widely in animal tissues and in microorganisms including soil bacteria. Its widespread distribution suggests that it has a rather fundamental function and environmental meaning. It has been shown previously that arylsulfatase of Klebsiella was purified and characterized. A condition of arylsulfatase synthesis was tested with several strains of Serratia. Serratia marcescens could not utilize some sugars, such as xylose, rhamnose, glucosamine and arabinose hut glucose and mannitol as a sole carbon source. However, arylsulfatase synthesis was repressed by glucose but not by mannitol. The enzyme synthesis was repressed ob inorganic sulfate and methionine, and this repression was relieved by addition of tyramine. Arylsulfatase of S. marcescen was purified by fractionation with ammonium sulfate and followed by chromatographies on DEAE-Cellulose CM-Cellulose, and DEAE-Sephadex A-25. The molecular weight of arylsulfatase was determined to be 46,000 by SDS-Gel electrophoresis and 49,000 by Sephadex G-100 column chromatography. The enzyme showed some different properties with that of K. aerogenes. The activity was maximum at pH 6.8. The Km and Vmax values for p-nitrophenyl sulfate were 2.5${\times}$10$\^$－4/ M and 20 nmoles/min/mg protein, respectively. The enzyme showed high activities toward phenyl sulfate, ο-and p-nitro phenyl sulfates, and p-nitrocatechol sulfate. The inhibition of enzyme was strongly affected by hydroxylamine, inorganic fluoride, sulfide and phosphate, but by inorganic sulfate. Like Klebsiella arylsulfatase, tyramine, octopamine, and dopamine gave signifcant inhibitory effect.

• Journal of Life Science
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• v.8 no.5
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• pp.614-621
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• 1998

An extracellular inulinase from Penicillium sp. which isolated from soil sample was purified to a single protein th-rough ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography and Toyopearl HW 65 F gel filtration. The temperature and pH for the enzyme reaction were around 6$0^{\circ}C$ and 4.0, respectively. The enzyme was stable at 3$0^{\circ}C$-5$0^{\circ}C$ and in the pH range of 4 to 5. $CuCl_2$, $HgCl_2$ and EDTA inhibited the enzyme activity strongly. By contrast, $MnCl_2$ and $CaCl_2$ activated the enzyme activity. The molecular weight of the purified enzyme was esti-mated to be 77,000 dalton by SDS-polyacrylamide gel electrophoresis. The Km values of the enzyme for inulin were calculated to be $2.2\times10^{-3}$M. TLC analysis suggested that purified enzyme is exo-type inulinase. The NH2-terminal amino acid sequences of the purified enzyme was determined to be $NH_2$-X-Glu-Ser-Tyr-Thr-Glu-Lys/Leu-Tyr-Arg-Pro.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.593-596
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• 1997

In the course of search antagonistic fungi from soil in green house, four kind of fungi (AF1, AF2, AF3, AF4) were isolated, which have activities against Phytophthora capsici, Botrytis cinera, Rhizoctonia solani, Pythium ultimum and Fusarium oxysporum. The AF2 was identified according to the morphological description of Aspergillus terreus. This antagonistic fungus inhibiting various plant pathogens was effective to reduce disease incidence of cucumber seedlings caused by mixed inoculum of Rhizoctonia solani, Pythium ultimum and Fusarium oxysporum. Antifungal compound I was isolated and purified by fresh chromatography from A. terreus. The $^1H$ and $^{13}C$ assignment of compound I was achieved from two-dimensional $^1H-^1H\;COSY$, HMQC, HMBC with the add of homonuclear and heteronuclear double resonance experiment. The compound I was identified butyrolactone I (${\alpha}$-oxo-${\beta}$-(p-hydroxyphenyl)-${\gamma}$-(p-hydroxy-m-3,3-dimethyl-allylbenzyl)-${\gamma}$-methoxycarbonyl-${\gamma}$-butyrolactone, $C_{24}H_{24}O_7$, M.W.=424).