• KSCE Journal of Civil and Environmental Engineering Research
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• v.35 no.4
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• pp.791-799
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• 2015

This study focuses on the algae growth restraining. Many researches on a critical damage from algae growth are published, but it is hard to find how th restrain. Abnormal algae increasing is a problem, because it makes red tides, biodeterioration, etc. Therefore this study aims to decrease the damage from algae growth. Some metal ions have been used microorganism killing materials from old times. Especially, Cu ions are highly effective. Based on these uses of the metal ions, a functional mortar which restrains algae growth is developed. The mortar contains soluble glass which dissolve in water. The soluble glass was made of Cu ions and phosphates. When the soluble glass is dissolved, Cu ions are soaked out stably from the soluble glass. Culture mediums which incubate algae were made to evaluate the developed mortar specimens. Culture mediums were filled with fresh water and sea water. Algae were incubated for fourteen days in culture mediums. The evaluating methods are measuring volume of the dissolved organic carbon and the chlorophyll. Using these two measurements, the mortar specimens are judged that can restrain algae or not. According to the result, the functional mortars of culture medium filled with fresh and sea water shows similar trend. The functional mortar for restraining algae growth performs that's role well.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.230-237
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• 2004

In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{＋}$ , $Cu^{ 2+}$, $Hg^{2＋}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2＋}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.131-137
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• 1998

The cellulase components were partially purified from the culture filtrate of the alkalophilic bacterium Pseudomonas sp. AC-711 and its enzymatic properties were characterized. The specific activity of the purified major enzyme component was 3.5 units/mg protein as carboxymethyl cellulase and the yield was 23% of the total activity of the culture broth. The molecular weight of the component was 46,000 and the Km and Vmax on CMC were determined as $15.4mg\;mL^{-1}$ and $4.17{\mu}moles\;mL^{-1}\;min^{-1}$, respectively. The enzyme was stable at the temperatures below $60^{\circ}C$ and at the pH range of 4.0~11.0, and the optimal temperature and pH were $60^{\circ}C$ and pH 8.0, respectively. The enzyme activity was not significantly affected by the common surfactants (concentration: 0.05%) such as ${\alpha}$-olefin sulfonate, linear alkylbenzene sulfonate, sodium dodecyl sulfonate, hexadecyltrimethylammonium bromide and Tween 80. The enzyme was activated by the metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Co^{2+}$, whereas inhibited by $Hg^{2+}$ and $Zn^{2+}$. The enzyme exhibited relatively high activity toward amorphous CMC as compared with crystalline substrates such as filter paper and avicel.

• Korean Journal of Microbiology
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• v.39 no.4
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• pp.223-229
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• 2003

The purpose of this work was to perform the characterization of NAD(P)H-nitroreductase isolated from Stenotrophomonas sp. OK-5 capable of degrading 2,4,6-trinitrotoluene (TNT). Initially, NADP(H)-nitroreductase by a series of purification processes including ammonium sulfate precipitation, DEAE-sepharose, andQ-sepharose was prepared. From samples harvested from fraction collector, three different fractions (I, II & III)having the enzyme activity of NAD(P)H-itroreductase were detected. Specific activities of three fractions I, II,and III of NAD(P)H-nitroreductase were determined to approximately 5.06 unit/mg, 4.95 unit/mg and 4.86 unit/mg, and concentrated to 10.5, 9.8, and 8.9-fold compared to crude extract, respectively. Among these three fractions,the fraction I of NAD(P)H-nitroreductase demonstrated the highest specific activity in this experiment. Several factors affecting on the enzyme activity of NAD(P)H-nitroreductase (fractions I, II & III) were investigated.The optimum temperature of all NAD(P)H-nitroreductase (fractions I, II & III) was 30oC, and the optimal pH was approximately 7.5. Metal ions such as Ag+, Cu2+, Hg2+ inhibited approximately 80% enzyme activity of all NAD(P)H-nitroreductase, and the enzyme activities were decreased about 30-40% inhibition in the presence of Mn2+ or Ca2+. However, Fe3+ showed stimulatory effect on the enzyme activity. The molecular weights of NAD(P)H-nitroreductase (fractions I, II & III) were measured about 27 kDa on the SDS-PAGE.

• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.142-150
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• 2002

Streptomyces polychromogenes IFO 13072 was used as a strain producing cholesterol oxidase(EC 1.1.3.6). The conditions of cholesterol oxidase production were investigated. The optimum composition of medium for production of the enzyme was 1% dextrin, 0.5% casamino acid, 0.1% $KH_2$PO$_4$, 0.5% $NaNO_3$ and 0.05% $MgSO_4$(pH 7.3). The enzyme was purified specifically by cholesterol affinity column chromatography with a yield of 23.2%. The purified enzyme showed a single polypeptide on SDS-PAGE and the molecular weight was estimated about 52,000 daltons. The optimum pH and temperature of the cholesterol oxidase were pH 7.0 and $37^{\circ}C$, respectively. The enzyme was stable in the range of pH 6.0~7.0 and $25^{\circ}C$. The cholesterol oxidase activity was strongly inhibited by metal ions such as $Hg^{2＋}$ and $Fe^{2＋}$ and inhibitors such as dithiothreitol, mercaptoethanol and isonicotinic acid. The Michaelis constant(Km) for the cholesterol was found to be 25 mM by Lineweaver-Burk plot analysis.

• Journal of Industrial Technology
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• v.39 no.1
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• pp.15-19
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• 2019

This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.4
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• pp.458-464
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• 1993

In the present paper enzymatic properties of the trypsins from the four dark fleshed fish were compared with each other and thermal stabilities of the enzymes were also investigated. The trypsins from the dark fleshed fish showed their activity only in BA-p-NA substrate of the amide substrates such as BA-p-NA and SP-p-NA, and BAEE and TAME of the ester substrate such as ATEE, BAEE, BTEE, and TAME. The enzymes were strongly inhibited by the serine protease inhibitors such as antipain, leupeptin, TLCK, DFP and SBTI, and were also inhibited by such metal ions as Cu$^{2＋}$ and Hg$^{2＋}$, but fairly activated by $Mg^{2＋}$. Denaturation constants of the enzymes were 13.4$\times$10$^{-4}$ sec$^{-1}$ for anchovy trypsin, 47.18$\times$10$^{-4}$ sec$^{-1}$ for mackerel trypsin A, 34.06$\times$10$^{-4}$ sec$^{-1}$ mackerel trypsin B, 42.28$\times$10$^{-4}$ sec$^{-1}$ for yellowfin tuna trypsin and 16.6$\times$10$^{-4}$ sec$^{-1}$ for albacore trypsin at 55$^{\circ}C$. The activation energies of the trypsins at a temperature range of 3$0^{\circ}C$ to 5$0^{\circ}C$ were estimated to be 13.91 ㎉/mole for anchovy trypsin, 11.61㎉/mo1e and 8.43㎉/mole for mackerel trypsin A and for mackerel typsin B, 4.35㎉/mole for yellowfin tuna trypsin, and 3.76㎉/mole for albacore trypsin.

• 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.

• Journal of Microbiology and Biotechnology
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• v.21 no.3
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• pp.256-262
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• 2011

A 20.9-kDa metalloprotease was isolated from dried fruiting bodies of the wild basidiomycete mushroom Lepista nuda. The N-terminal amino acid sequence of the protease was seen to be ATFVLTAATNTLFTA, thus displaying no similarity with the sequences of previously reported metalloproteases. The protease was purified using a procedure that entailed ion-exchange chromatography on CM-Cellulose, Q-Sepharose, and Mono S, and FPLC-gel filtration on Superdex 75. The protease functioned at an optimum pH of 7.0 and an optimum temperature of $50^{\circ}C$. It was also noted that the protease demonstrated a proteolytic activity of 1,756 U/mg toward casein. The $K_m$ of the purified protease toward casein was 6.36 mg/ml at a pH of 7.0 and with a temperature of $37^{\circ}C$, whereas the $V_{max}$ was 9.11 ${\mu}g\;ml^{-1}\;min^{-1}$. The activity of the protease was adversely affected by EDTA-2Na, suggesting that it is a metalloprotease. PMSF, EGTA, aprotinin, and leupeptin exerted no striking inhibitory effect. The activity of the protease was enhanced by $Fe^{2+}$, but was curtailed by $Cd^{2+}$, $Cu^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Fe^{2+}$ ions. The protease also exhibited inhibitory activity against HIV-1 reverse transcriptase with an $IC_{50}$ value of 4.00 ${\mu}M$. The $IC_{50}$ values toward hepatoma Hep G2 and leukemia L1210 cells in vitro were 4.99 ${\mu}M$ and 3.67 ${\mu}M$, respectively.

• Bulletin of the Korean Chemical Society
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• v.26 no.10
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• pp.1529-1532
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• 2005

A simple and cost effective method for separation and preconcentration of Ag(I) at the $10^{-7}\;mol\;L^{-1}$ level in the environmental and mineral samples is present. The method is based on the flotation of Ag(I)-iodide complex as an ion-associate with ferroin in pH of 4 from a large volume of an aqueous solution (500 mL) using nheptane. The floated layer was then dissolved in dimethylsulfoxide (DMSO) for the subsequent spectrophotometric determination. Beer's law was obeyed over a range of 2.0 ${\times}$ $10^{-7}$-4.0 ${\times}$ $10^{-6}$ mol $L^{-1}$ with the apparent molar absorptivity of 2.67 ${\times}$ $10^5$ L $mol^{-1}\;cm^{-1}$. The detection limit (n = 5) was 4 ${\times}$ $10^{-8}$ mol $L^{-1}$, and RSD (n = 5) obtained for 2.0 ${\times}$ $10^{-6}$ mol $L^{-1}$ of Ag(I) was 2.2%. The interference effects of a number of elements was studied and found that only $Hg^{2+}$ at low concentration, and $Pb^{2+}$, $Cd^{2+}$, $Cu^{2+}$, and $Fe^{3+}$ ions at moderately high concentrations were interfered. To overcome on these interference effects, the solution was treated with EDTA at a buffering pH of 4 and passed through a column containing Amberlite IR-120 ionexchanger resin, just before the flotation process. The proposed method was applied to determine of Ag(I) in a synthetic waste water, a photographic washing sample and a geological sample and the results was compared with those obtained from the flame atomic absorption spectrometry. The results were satisfactorily comparable with together, so that the applicability of the proposed method was confirmed in encountering with the real samples.