• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.427-434
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• 1998

An alkaline protease was 4-fold purified, yielding 2.3% of recovery by ammonium sulfate precipitation, CM-cellulose column chromatography and Sephadex G-100 column chromatography. The purified enzyme was estimated to be monomeric with molecular weight of about 62,000 from polyacrylamide gel eletrophoresis (PAGE) and sodiumdodecylsulfate polyacrylamide gel electrophoresis (SDS-FAGE). The optimal pH and temperature of the alkaline pretense activity were 11.0 and 50$^{\circ}C$, respectively, exhibiting high stability at pH value from 6.0 to 11.0 at 50$^{\circ}C$ for 30 minute. The alkaline pretense was activated by MnSO$_4$, CaCl$_2$, and was inhibited by CuSO$_4$, ZnSO$_4$, HgCl$_2$, EDTA and EGTA. Also, the enzyme was found to be a metaloenzyme requiring Mn$\^$2+/ as cofactor. The NH$_2$-terminal amino acid of alkaline protease was alanine. The Km and Vmax values of this enzyme for casein was 4.0 mg/$m\ell$ and 5,500 unit/$m\ell$, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.10
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• pp.1469-1473
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• 2011

The efficacy of antimicrobial agents and heat treatments on spore inactivation was investigated. Grapefruit seed extract (GFE) and ethylenediaminetetraacetic acid (EDTA) were used and as antimicrobial agents, and heat treatments were conducted at $70^{\circ}C$, $80^{\circ}C$, and $90^{\circ}C$ for 30 minutes. Heat treatments at $90^{\circ}C$ were the most effective on spore inactivation as a single treatment and caused a 2.3 log reduction. When combined with a single treatment to discover synergistic effects, 1% GFE with $80^{\circ}C$ heat treatments and 0.5 mM EDTA with $80^{\circ}C$ heat treatments resulted in 2.1 log and 3.2 log reductions, respectively, though they did not show reductions at each single treatment (GFE 1% (v/v), EDTA 0.5 mM, $80^{\circ}C$). So it was concluded that by combining GFE, EDTA in low concentration treatment, and heat treatment, B. cereus spores can be effectively inactivated.

• The Korean Journal of Food And Nutrition
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• v.10 no.3
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• pp.325-329
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• 1997

A inulinase of garlic(Seosan) was partially purified by ammonium sulfate fractionation and Sephadex G-150 gel filtration chromatography with a recovery of 9.1%. Optimum temperature and pH of the enzyme were 4$0^{\circ}C$ and pH 6.0, respectively, and the enzyme was stable below 7$0^{\circ}C$ and in the pH range of 5.0~8.0. The enzyme was strongly inhibited by metal ions(Al3+, Mn2+, Hg2+, Cd2+) and EDTA, and the Km value for inulin was 0.22%.

• Journal of Microbiology
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• v.43 no.3
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• pp.237-243
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• 2005

The protease purified from Bacillus cereus JH108 has the function of leucine specific endopeptidase. When measured by hydrolysis of synthetic substrate (N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide), the enzyme activity exhibited optimal activity at pH 9.0, $60^{\circ}C$. The endopeptidase activity was stimulated by $Ca^{++},\;Co^{++},\;Mn^{++},\;Mg^{++},\;and\;Ni^{++}$, and was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline, and EGTA. Addition of serine protease inhibitor, PMSF, resulted in the elimination of the activity. The endopeptidase activity was fully recovered from the inhibition of EDTA by the addition of 1 mM $Ca^{++}$, and was partially restored by $Co^{++}\;and\;Mn^{++}$, indicating that the enzyme was stabilized and activated by divalent cations and has a serine residue at the active site. Addition of $Ca^{++}$ increased the pH and heat stability of endopeptidase activity. These results show that endopeptidase requires calcium ions for activity and/or stability. A Lineweaver-Burk plot analysis indicated that the $K_m$ value of endopeptidase is 0.315 mM and $V_{max}$ is 0.222 ) is $0.222\;{\mu}mol$ of N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide per min. Bestatin was shown to act as a competitive inhibitor to the endopeptidase activity.

• Journal of the Korean Chemical Society
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• v.9 no.2
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• pp.101-105
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• 1965

Spectrophotometric method for the determination of Co(Ⅱ) is developed based on the fact that Co(Ⅱ) forms a stable red complex with 7-nitroso-8-hydroxyquinoline-5-sulfonate at pH 4. 5. The absorbance is measured at 528$m{\mu}$, $25^{\circ}C$. Beer's law is followed in the concentration range of 0. 3 to 6. 0 p.p.m. of Co(Ⅱ) and molar extinction coefficient of the complex was $1.1{\times}10^4$. Of the diverse ions checked, Fe(Ⅱ), Fe(Ⅲ), Cu(Ⅱ), Mn(Ⅱ), Hg(Ⅰ), CN-, EDTA interfere. The composition of the complex is found to be 3:1 ligand to metal species by mole ratio and continuous variation methods.

• KSBB Journal
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• v.5 no.2
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• pp.119-124
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• 1990

Phospholipase D catalyzes the phosphatidohydrolysis and transphosphatidylation of phospholipid in the biological systems. In this study we were partially purified phospholipase D from Chinese cabbage and the characterization of the enzyme was carried out in a multistirring batch system bioreactor. The enzyme showed optimum activity at pH ,5.6, highest activity at 37$^{\circ}C$ and Ca2+ is important for the enzyme activity. Optimum concentrations of Ca2+ for phosphatidohydrolysis was 20 mM and for transphosphatidylation was 40 mM, respectively. Some organic solvents such as diethylether, isopropylether and butylacetate were activated the enzyme activity. On the other hand, EDTA, Ba2+, Mn2+ and Zn2+ showed inhibitory effect on the enzyme activity. The base acceptors in transphosphatidylation by the Chinese cabbage phospholipase D were tested. Various poly-and monohydroxy alcohols were found to be active.

• Journal of the Korean institute of surface engineering
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• v.36 no.5
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• pp.379-385
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• 2003

The surface morphology, the glossiness and the hardness of Zn-Cr and Zn-Cr-X(X:Co, Mn) alloy electrodeposits were investigated by using chloride bath with EDTA additive and flow cell system. The surface morphology of Zn-Cr alloy and Zn-Cr-Mn alloy changed from fine needle shape crystalline structure to colony structure of fine granular crystallites with increasing current density in the range of 20-100 $A/dm^2$. The surface morphology of Zn-Cr-Co alloy deposited from low Co concentration bath(2.5-10 g/$\ell$) was similar to that of Zn-Cr alloy, while that of Zn-Cr-Co alloy deposited from high cobalt concentration bath was fine granular crystalline structure in the same range of current density. The glossiness of Zn-Cr and Zn-Cr-Mn alloy increased noticeably with increasing current density, while that of Zn-Cr-Mn alloy decreased with increasing Mn concentration of bath in high current density region. The glossiness of Zn-Cr-Co alloy deposited from low Co concentration bath increased with current density while that of the alloy from high Co concentration bath decreased with increasing current density. The hardness of Zn-Cr and Zn-Cr-X alloy increased noticeably with current density.

• Journal of the Korean institute of surface engineering
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• v.36 no.3
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• pp.256-262
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• 2003

The current efficiency and the composition of Zn-Cr and Zn-Cr-X (X : Co, Mn) alloy electrodeposits were investigated by using chloride bath with EDTA auditive and flow cell plating system. The current efficiency of Zn-Cr alloy decreased with increasing current density, while it increased with the content of Co and Mn of the Zn-Cr-X alloy bath in high current density region. The Cr content in Zn-Cr alloy increased from 1.4-2.7 to $28wt\%$ with increasing current density and the phase structure of the alloys changed from $\eta-Zn$ through $\eta-Zn+\gamma'-ZnCr\;to\;\gamma'-ZnCr$ with Increasing Cr content of the alloys. The Co content in Zn-Cr-Co alloys increased with Co content of the bath, while Cr content of the alloy increased or decreased in low current density region $(10-75A/dm^2)$ or high current density region $(75-100A/dm^2)$, respectively. $\gamma-ZnCo$ phase was formed in the Zn-Cr-Co alloy with above $9.0wt\%$ Co. The content of Mn and Cr in Zn-Cr-Mn alloys increased or decreased with the increase of current density in high current density region, respectively while Cr content of the alloy decreased noticeably with the increase of Mn content in the bath. Two phases of $\delta_1-ZnMn$ and $\gamma'-ZnCr$ were formed in the Zn-Cr-Mn alloy with above $8.6wt\%$ Mn.

• The Korean Journal of Mycology
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• v.24 no.1 s.76
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• pp.8-16
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• 1996

The ${\alpha}-amylase$ of Alternaria alternata was purified through ammonium sulfate precipitation, dialysis and Sephadex G-100 column chromatography. One single band was obtained in SDS-polyacrylamide gel electrophoresis. The optimum pH for enzyme activity was 5.0 and the enzyme activity was maintained at $3.6{\sim}7.0$pH range. The optimum temperature for ${\alpha}-amylase$ activity was $40^{\circ}C$ and 71% of the activity was still maintained until 30 min after heating at $80^{\circ}C$. The ${\alpha}-amylase$ was slightly activated by $Mn^{2+},\;Zn^{2+}\;and\;Sn^{2+}$, but inhibited by $Ba^{2+},\;Pb^{2+},\;Co^{2+}\;and\;Ag^{1+}$. The $Hg^{2+}\;and\;Ag^{2+}$ slightly inhibited the activity of the enzyme at concentrations of $10^{-3}\;and\;10^{-4}M$. The Michaelis constant $(K_m)$ to soluble starch was $6.50{\times}10^{-2}M$ and inhibition constant $(K_i)$ by the 1mM EDTA was $8.0{\times}10^{-2}M$. The inhibition of this enzyme by EDTA was competitive one.

• Korean Journal of Food Science and Technology
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• v.12 no.1
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• pp.13-17
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• 1980

A Streptomyces spp. strain SY 79-1 which was capable of producing metal protease was isolated from soil. The optimal pH and temperature of the protease were around pH 8.0 and $45^{\circ}C$, respectively. The stable pH range of the enzyme was between pH 6.0 to 8.0. The enzyme was stable at $45^{\circ}C$, but it lost the activity about 75 % for 5 min and completely for 30 min when it was treated at $60^{\circ}C$. The activity of the enzyme was inhibited by $Hg^{++},\;Cu^{++},\;Ag^{+}$ and activated by $Mg^{++},\;Mn^{++},\;Co^{++},\;but\;Fe^{++},\;Ca^{++},\;Pb^{++}\;and\;Al^{3+}$ did not affect enzyme activity. This enzyme was strongly inhibited by EDTA, but was not inhibited by 2, 4-DNP, ${\rho}$-CMB, ${\varepsilon}$-aminocaproic acid, cysteine, thiourea, citric acid, oxalic acid and sodium arsenate. When cobalt was added to the EDTA-denatured enzyme, the activity of the enzyme was restored.