• Journal of Korean Society of Environmental Engineers
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• v.22 no.3
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• pp.475-484
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• 2000

The purpose of this study was to develop model for estimating biodegrability of organic sludge (sewage and papermill) in various environmental conditions. to assume degradable degree with operating time of SRB reactor. and evaluate duratior of organic sludge as carbon source. Average TCOD was 28.7~63.2mg/L in effluent. organic sludge did not much supply carbon source for experimental period. But in point of durability. it seemed that organic sludge was efficient because it was not consumed by degradation of much organic matter within short period. With increasing $SO_4{^{2-}}$ reduction rate. Pb and Fe was removed 77~82% and 33~59%. respectively. Because Al was precipitated as a hydroxide. its removal rate wa,. about $54{\pm}2%$ in R-l~R-3 maintaining low pH but about 78% in R-4 maintaining high pH. Because Mn was large in solubility. it showed to be much lower than other heavy metals. Considering supportable capacity or durability of orgainc matter for initial SRB mixing ratio of sewage/papermill 0.5 was regarded as appropriate substituting material and at this time. it estimated that carbon source continued about 3.08 year but safety factor must apply to be thought over. because various factors had an effect on degradation of organic sludge.

• The Korean Journal of Pesticide Science
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• v.12 no.4
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• pp.315-322
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• 2008

In this study, a simple, effective, and sensitive method has been developed for the quantitative residue analysis of cyhalofop-butyl and its metabolite cyhalofop acid in water and soil when kept under laboratory conditions. The content of cyholofop-butyl and cyhalofop acid in water and soil was analyzed by first purifying the compounds through liquid-liquid extraction and partitioning followed by Silica gel (adsorption) chromatography. Upon the completion of the purification step the residual levels were monitored through high-performance liquid chromatography (HPLC) using a UV absorbance detector. The recoveries of cyhalofop-butyl from three replicates spiked at two different concentrations ranged from 82.5 to 100.0% and from 66.7 to 97.9% in water and soil, respectively. The limit of detection and minimum detection level of cyhalofop-butyl in water and soil was 0.02 ppm and 10 ng, respectively. The recoveries of cyhalofop acid ranged from 80.7 to 104.8% in water and from 76.9 to 98.1 % in soil. The limit of detection of cyhalofop acid was 0.005 ppm in water and 0.01 ppm in soil, while the minimum detection level was 2 ng both in water and soil. The half-live of cyhalofop-butyl was 4.14 and 6.6 days in water and soil, respectively. The method was successfully applied to evaluate cyhalofop-butyl residues in water and soil applied aj. 30% emulsion, oil in water (EW) product.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.315-321
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• 1994

Glyoxalase I was purified 2,294-fold from Pleurotus ostreatus by S-hexylglutathione affinity chromatography, Sephadex G-150 gel filtration chromatography and DEAE-sepharose A-50 CL-6B ion exchange chromatography with an overall yield of 21.7%. The molecular mass determined by gel filtration was found to be approx. 34 kDa. SDS-PAGE revealed that the enzyme consists of two identical subunits with a molecular mass of approx. 17 kDa. The K sub(m) values of this enzyme for methylglyoxal and phenylglyoxal were 0.39 mM and 0.22 mM, respectively. And this enzyme had a strong affinity for L-xylosone and hydroxypyruvaldehyde. The enzyme showed its optimal activity at pH 6.5-7.5 and at $40^{\circ}C$. $^1H$-NMR spectroscopic analysis of enzymic reaction showed that this enzyme catalyzes intramolecular proton transfer.

• Bulletin of the Korean Chemical Society
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• v.33 no.10
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• pp.3248-3252
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• 2012

Vibrio extracellular metalloprotease (vEP), secreted from Vibrio vulnificus, shows various proteolytic function such as prothrombin activation and fibrinolytic activities. Premature form of vEP has an N-terminal (nPP) and a C-terminal (C-ter100) region. The nPP and C-ter100 regions are autocleaved for the matured metalloprotease activity. It has been proposed that two regions play a key role in regulating enzymatic activity of vEP. Especially, C-ter100 has a regulatory function on proteolytic activity of vEP. C-ter100 domain has been cloned into the E. coli expression vectors, pET32a and pGEX 4T-1 with TEV protease cleavage site and purified using gel-filtration chromatography followed by affinity chromatography. To understand how C-ter100 modulates proteolytic activity of vEP, structural studies were performed by heteronuclar multi-dimensional NMR spectroscopy. Backbone $^1H$, $^{15}N$ and $^{13}C$ resonances were assigned by data from standard triple resonance and HCCH-TOCSY experiments. The secondary structures of vEP C-ter100 were determined by TALOS+ and CSI software based on hydrogen/deuterium exchange. NMR data show that C-ter100 of vEP forms a ${\beta}$-barrel structure consisting of eight ${\beta}$-strands.

• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.1
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• pp.33-44
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• 2003

The arylphorin was purified from the pupal haemolymph of the Chinese oak silkmoth, Antheraea pernyi, and characterized physiologically and biochemically, The protein was purified by a simple preparative polyacrylamide gel electrophoresis (PAGE) and subsequent diffusive elution. The preparation was shown to be homogeneous by 7.5% native-PAGE. The native molecular weight of arylphorin was 450 kDa with a 80 kDa single subunit, suggesting hexamer, The protein contained high amounts (18.3%) of aromatic amino acids, phenylalanine (9.7%) and tyrosine (8.6%). Therefore, the protein was identified as a kind of a storage protein referred to as an arylphorin. The protein was stained by Schiff's reagent, suggesting a glycoprotein. The protein contained 4.9% (w/w) sugar and mannose and N-acetylglucosamine were major components. Also, degradation of the protein was begun by heat treatment at 90 for 20 minutes. These results showed that the A. pernyi arylphorin in the study is hexamer associated with the six subunits consisting of a 80kDa single subunit, and is different from that of Kajiura et al. (1998) in the subunit composition.

• BMB Reports
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• v.33 no.2
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• pp.112-119
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• 2000

Three kinds of serine protease inhibitors, members of the Bowman-Birk trypsin inhibitor, were purified from Dolichos lablab seeds and named Dolichos protease inhibitor 1, 2 and 3 (DI-1, DI-2 and DI-3), respectively. Each inhibitor showed a single band with gel mobility at around 15.9, 12.1 and 14.6 kDa on 20% SDS-PAGE under reducing conditions. To characterize inhibitory specificity, the inhibition constant (Ki) for these inhibitors was measured against several known serine proteases. All three Dolichos protease inhibitors (DI-1, DI-2 and DI-3) inhibited the activity of trypsin and plasmin, but had no effect on thrombin and kallikrein (either for human plasma kallikrein or for porcine pancreas kallikrein). DI-1 inhibited chymotrypsin most effectively (Ki = $3.6{\times}10^{-9}\;M$), while DI-2 displayed inhibitory activity for porcine pancreatic elastase (Ki = $6.2{\times}10^{-8}\;M$). Pre-treatment of the 33 mg/kg of DI-mixture (active fractions from $C_{18}$ open column chromatography that included DI-1, DI-2 and DI-3) inhibited the induction of pseudomonal elastase-induced septic hypotension and prevented an increase in bradykinin generation in pseudomonal elastase-treated guinea pig plasma. Also, the increase of kallikrein activity, by injection of pseudomonal elastase, was inhibited by the pretreatment of the DI-mixture in a guinea pig. Since the DI-mixture had no inhibitory effect on kallikrein activity when Z-Phe-Arg-MCA was used as a substrate in vitro, its inhibitory activity in the pseudomonal elastase-induced septic hypotension model might not be due to a direct inhibition of plasma kallikrein in the activation cascade of the Hageman factor and prekallikrein system. These results suggest that the Dolichos DI-mixture might be used as an inhibitor in pathogenic bacterial protease-induced septic shock.

• Microbiology and Biotechnology Letters
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• v.46 no.3
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• pp.244-252
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• 2018

Protein disulfide isomerase A3 (PDIA3) is a major member of the protein disulfide isomerase (PDI) family. PDI proteins commonly reside in the endoplasmic reticulum and mediate important thiol-disulfide interchanges during post-translational protein folding. Unlike other PDI family members, PDIA3 is ubiquitous in various organ systems. However, its physiological activity varies in other tissues. PDIA3 has been associated with cancer, airway inflammation, neurodegenerative diseases, and metabolic diseases. However, the mechanisms of the association of PDIA3 with these pathological conditions remain unclear. Recombinant PDIA3 (rPDIA3) is needed to clarify the interactions between PDIA3 and certain physiological phenomena. In the present study, we aimed to produce highly purified rPDIA3 for use in pathological experiments. We expressed rPDIA3 with a histidine-enriched elongated peptide tag in Escherichia coli and obtained rPDIA3 at 97.8% purity using consecutive His-tag and reverse-phase chromatography. Elongated peptide tags screened from artificially designated library had dual functions for protein expression and simple purification.

• Journal of Microbiology and Biotechnology
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• v.1 no.3
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• pp.188-196
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• 1991

The ${\beta}-glucosidase$ was purified to homogeneity from the culture filtrate of P. verruculosum by column chromatography. The enzyme was a glycoprotein with a relative size of approximately 220 kDa with an isoelectric point of 4.8, which was composed of dimeric protein of 105 kDa. The enzyme was stable up to $60^{\circ}C$ and the presence of glycerol significantly increased its thermostability. The enzyme was found to hydrolyze both ${\beta}-aryl$ and ${\beta}-alkyl-glucosides$ in addition to ${\beta}-glucosyl$ glucose and catalyzed glucosyl transfer to cellobiose. The enzyme attacked laminarin in an exotype-like fashion. The apparent Km's of the enzyme toward cellobiose, laminaribiose, laminarin were 0.53 mM, 0.35 mM and 1.11 mM, respectively. Glucose and glucono-${\delta}-lactone$ were competitive inhibitors for the enzyme. Copper ($Cu^{2+}$), mercury ($Hg^{2+}$) and p-chloromercuribenzoate were strong inhibitors of the enzyme. The immunoblotting result revealed that one form of ${\beta}-glucosidase$ was biosynthesized, irrespective of carbon sources used. Polyacrylamide gel electrophoresis analysis of the in vitro translated product of total RNA from avicel grown mycelium established that the P. verruculosum ${\beta}-glucosidase$ precursor was approximately 95 kDa in size. The amino acid composition and N-terminal amino acid sequence are given.

• Journal of the East Asian Society of Dietary Life
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• v.4 no.3
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• pp.59-65
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• 1994

Growth-stimulating effects of cow's milk was examined using Vero cell cultre. Medium containing whole cow's milk stimulated cell growth about the same degree as that containing fetal bovine serum. The growth-stimulating factor in cow's milk was purified using hydrophobic (phenyl-sepharose) and gel filtration (Sephadex G-100) column chromatographies. It appeared that the factor is a highly hydrophobic protein, since the major growth-stimulating activity was found in the fractions eluted with 50% ethylene glycol from the phenyl-sepharose column during the purification. The purified factor showed a single band on the polyacrylamide gel electrophoresis in the presence of 1% (w/v) SDS. The factor has been found to have a relatively high molecular weight in the range of about Mr=100,000-150,000. In the presence of the purified factor (5%, w/v) in the culture medium, the incorporation of [3H]-thymidine into the cells was increased approximately 2,400-fold over that in the presence of 5% (w/v) fetal bovine serum. It seems that the growth-stimulating factor purified in this study is one of the major growth factors in the cow's milk.

• BMB Reports
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• v.36 no.2
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• pp.230-236
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• 2003

Hot water-soluble crude polysaccharide (HCAP-0) that was obtained from the fruits of Capsicum annuum showed potent anti-complementary activity. The activity was unchanged by pronase digestion, but decreased by periodate oxidation. The HCAP-0 was fractionated by DEAE ion-exchange chromatography to give two major fractions, HCAP-II and III. These two fractions were finally purified by gel filtration to give HCAP-IIa, HCAP-IIIa1, and IIIa2 fractions that had high anti-complementary activities. The HCAP-IIIa1 and IIIa2 consisted of homogeneous polysaccharides. The anti-complementary activities were unaffected by treatment with polymyxin B, indicating that the modes of complement activation were not due to preexisting lipopolysaccharide. The molecular weight and sugar content of HCAP-IIIa2 had potent anti-complementary activity. The highest yields were 55 kDa and 75.9%, and the molar ratio of galactose (Ara:Gal, 1.0:4.6) was higher than other sugars. The crossed immuno-electrophoresis showed that both classical and alternative pathways were activated by HCAP-IIIa2.