• Korean Journal of Microbiology
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• v.31 no.2
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• pp.175-178
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• 1993

Alkaline .alpha.-amylase expressed in the transformant, Baciollus subtills MB809, containing alkaline amylase gene cloned from alkalophilic Bacillus sp. AL-8, was purified through for step separation processes. The purified alkaline .alpha.-amylase had molecular weight of app[roximately 59, 000 daltons on SDS-PAGE and Sephaex G-100 gel filtration. Amino acid sequence of terminal portion of the enzyme was analyzed with pure amylase eluted form the SDS-PAGE gel. N-terminal amino acid sequence of .alpha.-amylase was determined by the Edman degradation method and resulted in $NH_{2}$-ser-thr-ala-pro-ser-(ile)-lys-ala-gly-thr-(ile)-leu. For C-terminal amino acid sequencing, purified .alpha.-amylase was digested with carboxypuptidase A and B, and reverse-phase HPLC gradient elution system resulted in -thr-trp-pro-lys-COOH.

• BMB Reports
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• v.30 no.4
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• pp.258-261
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• 1997

A simple quantitative assay method for ribonuclease activity has been developed. This method is based on the decrease of fluorescence intensity emitted by the ethidium bromide bound to RNA due to the degradation of RNA by ribonuclease. The substrate RNA was reacted with ribonuclease A and the fluorescence intensity was measured after the addition of ethidium bromide. The intensity difference was calculated using a blank reaction mixture containing no RNase. Whole cellular RNA substrate produced a significant error and was not suitable for this assay method possibly because of local microheterogeniety caused by high molecular weight rRNA. but satisfying results were obtained with tRNA substrate. The intensity difference increased linearly by raising enzyme concentration up to $2{\times}10^{-4}$ Kunitz Units of ribonuclease A. More refined and reliable results were obtained by use of initial reaction velocities which were calculated from the plots of intensity difference vs time. A linear relationship between initial velocities and enzyme concentrations was observed up to 0.01 Kunitz Units of enzyme.

• BMB Reports
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• v.34 no.5
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• pp.440-445
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• 2001

Pesticide waste and chemical stockpiles are posing a potential threat to both Vie environment and human health. There is currently a great effort toward developing effective and economical methods for the detoxification of these toxic organophosphates. In terms of safety and economy, enzymatic biodegradation has been recommended as the most promising tool to detoxify these toxic materials. To develop an enzymatic degradation method to detoxify such toxic organophosphorus compounds, a gene encoding organophosphorus acid anhydrolase (OPAA) from genomic DNA of Alteromonas haloplanktis C was subcloned and expressed. The enzyme consists of a single polypeptide chain with a molecular weight of 48 kDa. It demonstrates strong hydrolyzing activity on sarin, an acetylcholinesterase inhibitor. Moreover, its high activity is sustained for a considerable length of time. It is projected that the recombinant OPAA can be applied as an enzymatic tool that can be used not only for the detoxification of pesticide wastes, but also for the demilitarization of chemical stockpiles.

• Food Engineering Progress
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• v.13 no.3
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• pp.169-175
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• 2009

Water soluble polysaccharides (WSP) and arabinogalactan of Portulaca oleracea L. (POL) were increased after extrusion and commercial cellulase treatment. Arabinose and galactose content increased more about 1.5 times than those of raw POL, and rhamnose also increased about 2.6 times in WSP. High molecular weight fraction (I) of POL depending on extrusion condition including Ext I, Ext II and Ext III degraded into low molecular weight fraction (II) about 37, 29, and 26%, respectively, ranged from 67,000-69,000 Da of molecular weight. Especially, the molecular weight and composition of WSP with extruded, were increased from 9 to 13% in low molecular weight fraction, compared to those of raw POL. Solubilization and degradation of polysaccharides were a directly propotional to specific mechanical energy in POL extrusion. WSP obtained by extrusion at Ext I and Ext II were found to be effective antioxidants in different in vitro assays with regards to 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and Trolox equivalent antioxidant capacity (TEAC). However, these results suggest that WSP obtained using extrusion and subsequent enzymatic treatment may be an effective method to produce arabinogalactan from POL and be used as a functional food ingredients.

• Analytical Science and Technology
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• v.9 no.1
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• pp.1-12
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• 1996

The thermally labile compounds with hydroxyl group generate the [${MNH_4}^+$] ion, [$MH^+$] and [$MH^+-OH$] ion by ion-molecule reaction in LC/TSP/MS. But these ions create the trouble in the estimation of molecular weight of an unknown compound because the margin of [${MNH_4}^+$] ion and [$MH^+$] ion is same to that of [$MH^+$] ion and [$MH^+-OH$] ion. If it is compensated for the results by using of the $CF_3COOD+NH_4OH$ LC eluent, the molecular weight of analyte will be able to be confirmed. And this study was tried to recognize whether the fragment ions of thermally labile compound are generated by electron impact or by thermal degradation in ion source.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.515.1-515
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• 1986

Thermostable tryptophanase was extracted from a thermophilie bacterium, strain T which was absolutely symbiotic with strain 5. The enzyme was purified 14.7 fold with 5.8% yield by chromatographies using ion exchange, gel filtration, and hydrophobic interaction columns, followed by high performance liquid chromatography on hydroxyapatite column. The purified enzyme has a molecular weight of approximately 210,000 estimated by gel filtration column chromatography, and the molecular weight of subunit was determined by SDS polyacrylamide gel electrophoresis to be 46,000, which indicates that the native enzyme is made of four homologous subunits. The tryptophanase was stable at 65o0 and the optimum temperature for the enzyme activity for 20 min reaction was 70$^{\circ}C$. The purified enzyme activity for 20 min ieaction was 70$^{\circ}C$. The purified enzyme catalyzed the degradation of L-tryptophan into indole, pyruvate and ammonia in the presence of pyridoxal phosphate. 5-Hydroxy-Ltryptophan, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-L-cysteine, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-Lcysteine, and L-serine were also used as substrates to form pyruvate. The amino acid composition of the tryptophanase was determined, and found to contain a high percentage of hydrophobic amino acids, especially in the proline content, which was much higher than that of Escherichia coli tryptophanase. In addition, the 35N-terminal amino acid sequence of the tryptophanase was completely different from that of E. coli tryptophanase.

• Archives of Pharmacal Research
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• v.24 no.1
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• pp.69-73
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• 2001

Biodegradable hydrogels based on glycidyl methacrylate dextran (CMD) and dimethacrylate poly(ethylene glycol) (DMP) were proposed for colon-specific drug delivery. GMD was synthesized by coupling of glycidyl methacylate with dextran in the presence of 4-(N, N-dimethylamino)pyridine (DMAP) using dimethylsulfoxide as a solvent. Methacrylate-terminated poly (ethylene glycol) (PEG) macromer was prepared by the reaction of PEG with methacryloyl chloride. CMD/DMP hydrogels were prepared by radical polymerization of phosphate buffer solution (0.1 M, pH 7.4) of GMD and DMP using ammonium peroxydisulfate (APS) and UV as initiating system. The synthetic GMD, DMP and GMD/DMP hydrogels were characterized by fourier transform infrared (FT-lR) spectroscopy. The FITC-albumin loaded hydrogels were prepared by adding FITC-albumin solution before UV irradiation. Swelling capacity of GMD/DMP hydrogels was controlled not only by molecular weight of dextran, but also by incorporation ratio of DMP Degradation of the hydrogels has been studied in vitro with dextranase. FITC-albumin release from the GMD/DMP hydrogels was affected by molecular weight of nextran and the presence of dextranase in the release medium.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.3
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• pp.273-279
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• 1993

Hydrolytic patterns of 11S globulin (glycinin), storage protein of soybean, by soymilk-clotting enzymes Iand IIfrom Bacillus sp. K-295G-7, which was the first soymilk-clotting enzyme to be found in a bacteria, was investigated. The clotting time of about 4~5 min is revealed by the Enzymes Iand II(0.025 units at 35$^{\circ}C$) on the acidic subunit. In electrophoresis, acidic subunit (A$_3$, M.W. 45,000) disappeared almost completely within 2 min and new products corresponding to the molecular weight of 16,000 and 20,000 were formed by the action of Enzymes I and II. Furthermore, Enzyme II produced a degradation compound having a molecular weight of about 30,000. In contrast, the hydrolytic patterns of basic subunit (M.W. 20,000) by Enzymes I and II were similar, but Enzyme II produced low molecular weight products slower than that of Enzyme I.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.254-263
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• 2014

Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ${\alpha}$-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in high-level expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at $40^{\circ}C$. The ${\alpha}$-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ${\alpha}$-amylase can be useful in lowering EP molecular weight and in starch processing.

• Applied Chemistry for Engineering
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• v.10 no.3
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• pp.432-437
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• 1999

Pyrolysis of polyethylene was carried out in the stainless steel reactor of internal volume of $10cm^3$. Pyrolysis reactions were performed at temperature $390{\sim}450^{\circ}C$ and the pyrolysis products were collected separately as reaction products and gas products. The molecular weight distributions(MWDs) of each product were determined by HPLC-GPC and GC analysis. Distribution balance equation for MWDs of random and specific products were proposed to account for initiation-termination and propagation-depropagation, such as hydrogen abstraction, chain cleavage, coupling of polymer and radical. A separate chain-end scission process produces low molecular weight noncondensable gases(C1 through C5) of average molecular weight 38. Activation energies of the random-chain scission and chain-end scission rate parameters, respectively, were determined to be 35, 17 kcal/mole.