• Journal of Plant Biology
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• v.34 no.2
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• pp.121-127
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• 1991

Total soluble proteins from three developmental stages of induced root nodules of Canavalia lineata were compared with those of non-nodulated roots by SDS-PAGE and two dimensional (2-D) gel electrophoresis. Thirteen nodule-specific protein (nodulin) bands were identified by the former and 30 nodule specific protein spots were detected by the latter method respectively. Some of the nodulins were detected differentially depending on the nodule's developmental stages. For example, only three leghemoglobin (Lb)-like protein spots appeared at stage I (d<2 mm), but two additional Lb-like protein spots appeared at stage II (d <4-5 mm). pI value and molecular weight of nomomers of Lb-like protein were narrower and greater than those of soybean, ranging from 4.4 to 5.0 and 15.7 kd respectively. Northern blot hybridization of total RNAs from roots and root nodules using soybean Lb cDNA as a probe made it clear that Lb gene was expressed tissue-specifically only in the root nodules.odules.

• Proceedings of the Microbiological Society of Korea Conference
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• 2001.11a
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• pp.157-163
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• 2001

Mycolic acid-containing gram-positive bacteria, so called nocardioform actinomycetes, have become a great interest to environmental microbiologists due to their metabolic versatility, multidegradative capacity and potential for bioremediation of priority pollutants. For example, Rhodococcus rhodochrous N75 was able to metabolize 4-methy1catechol via a modified $\beta$-ketoadipate pathway whereby 4-methylmuconolactone methyl isomerase catalyzes the conversion of 4-methylmuconolactone to 3-methylmuconolactone in order to circumvent the accumulation of the 'dead-end' metabolite, 4-methylmuconolactone. R. rhodochrous N75 has also shown the ability to transform a range of alkyl-substituted catechols to the corresponding muconolactones. A novel 3-methylmuconolactone-CoAsynthetase was found to be involved in the degradation of 3-methylmuconolactone, which is not mediated in a manner analogous to the classical $\beta$-ketoadipate pathway but activated by the addition of CoA prior to hydrolysis of lactone ring, suggesting that the degradative pathway for methylaromatic compounds by gram-positive bacteria diverges from that of proteobacteria. Mycobacterium sp. Strain PYR-l isolated from oil-contaminated soil was capable of mineralizing various polyaromatic hydrocarbons (PAHs), such as naphthalene, phenanthrene, pyrene, fluoranthrene, 1-nitropyrene, and 6-nitrochrysene. The pathways for degradation of PAHs by this organism have been elucidated through the isolation and characterization of chemical intermediates. 2-D gel electrophoresis of PAH-induced proteins enabled the cloning of the dioxygenase system containing a dehydrogenase, the dioxygenase small ($\beta$)-subunit, and the dioxygenase large ($\alpha$)-subunit. Phylogenetic analysis showed that the large a subunit did not cluster with most of the known sequences except for three newly described a subunits of dioxygenases from Rhodococcus spp. and Nocardioides spp. 2-D gel analysis also showed that catalase-peroxidase, which was induced with pyrene, plays a role in the PAH metabolism. The survival and performance of these bacteria raised the possibility that they can be excellent candidates for bioremediation purposes.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.32-32
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• 2003

소 난자의 체외성숙 과정에서 세포질 내 단백질의 생산과 축적의 변화는 핵 및 세포질 성숙과 밀접한 관계가 있다는 보고가 있지만, 난자의 성숙과 관련된 특정 단백질의 종류에 대한 보고는 없었다. 따라서 본 연구는 한우 난자의 체외성숙과 관련된 단백질의 생산 및 축적의 변화와 그 종류에 대해서 검토하였다. 체외성숙 시간(4.5, 9, 13.5, 18 및 24시간)에 따른 배양액 내의 단백질 합성의 변화는 2D gel electrophoresis를 이용하였고, 단백질 spot에 대해서는 peptide mass fingerprinting(PMF) 방법을 이용하였다. 또한 단백질 측정 시간에 신선 체외성숙 배양액으로 교환 후 난포란의 핵성숙과 배발달율을 검토하였다. 그 결과 한우 난포란의 체외성숙 시간에 따라 배양액에서 단백질의 양 및 질적인 변화를 확인하였다. 그리고 총 296개 단백질 spot들을 확인하였고, 그 중 30개 spot에서 유의적인 변화가 인정되었다. 또한 유의적인 변화를 보인 spot에 대한 PMF 분석을 통하여 Apolipoprotein A-1 precursor, Alpha enolase, Aldose reductase, 43kDa collectin precursor, Heat shock 27kDa protein, Plasminogen activator inhibitor-1 precursor, Thrombospondin 1 및 Transitional endoplasmic reticulum ATPase가 동정되었다 그리고 총 단백질 합성 경향은 0∼4.5 시간에는 감소하였고, 13.5∼18시간에 증가 한 후 다시 감소하는 경향을 나타내었으며, 단백질의 종류도 시간대별로 현저한 변화가 있었다. 한편 단백질을 측정하는 시기에 신선 체외성숙 배양액으로 교환한 후 난포란의 핵성숙 및 배발달율을 검토한 결과 18시간 체외성숙군에서 9시간째의 교환이 유의적으로 높은 핵성숙을 나타내었으나, 배발달율에서는 유의성이 인정되지 않았다. 그러나 24시간 체외성숙군에서 18시간째의 배양액 교환은 8세포기 및 배반포 발달율이 유의적(P<0.05)으로 높았다. 연구 결과로부터 소 난자의 체외성숙 시간에 따른 단백질 합성 경향의 차이를 확인하였고, 유의적인 변화를 나타낸 8가지의 단백질을 분리할 수 있었으며, 향후 이들 작용기전에 대한 연구가 필요하다고 사료된다.

• Parasites, Hosts and Diseases
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• v.26 no.2
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• pp.73-86
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• 1988

The sensitivity and specificity of crude and affinity-purified antigens of Clcnorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent astray (ELISA). The results were as follows: 1. The antibody.binding antigen (ABA) purified from whole worm crude antigen (IVWA) by CNBr-activated Sepharose 4B affinity chromatography made :l specific bands against rabbit antisera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16, 300~18, 500 and 28, 000~29, 000 daltons, while major ABA protein bands were at 18, 000~21, 000 and 29, 000~31, 000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. 2. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15, 000-200, 000 daltons, 15, 000-100, 000 daltons and 11, 000~80, 000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31, 000 daltons, while those of EGA and MEA consisted of higher molecular T.eights than 30, 000 daltons. 3. The ELISA reactivities of WWA to rabbit anti.sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4~6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting witll WWA showed a plateau without variation. MEA shoT.ed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly in(ected groups throughout the experiments, althougll there were some wealth positive cases (O.D.>0.6) ill heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential 18 increase the sensitivity and specificity for the immuncdiagnosis of clonorchiasis.

• The KIPS Transactions:PartB
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• v.15B no.6
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• pp.561-574
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• 2008

The spot detection phase of the 2-DE image analysis program segments a gel image into spot regions by an image segmentation algorithm and fits the spot regions to a spot shape model and quantifies the spot informations for the next phases. Currently the watershed algorithm is generally used as the segmentation algorithm and there are the Gaussian model and the diffusion model for the shape model. The diffusion model is closer to real spot shapes than the Gaussian model however spots have very various shapes and especially an asymmetric formation in x-coordinate and y-coordinate. The reason for asymmetric formation of spots is known that a protein could not be diffused completely because the 2-DE could not be processed under the ideal environment usually. Accordingly we propose an asymmetric diffusion model in this paper. The asymmetric diffusion model assumes that a protein spot is diffused from a disc at initial time of diffusing process, but is diffused asymmetrically for x-axis and y-axis respectively as time goes on. In experiments we processed spot matching for 19 gel images by using three models respectively and evaluated averages of SNR for comparing three models. As averages of SNR we got 14.22dB for the Gaussian model, 20.72dB for the diffusion model and 22.85dB for the asymmetric diffusion model. By experimental results we could confirm the asymmetric diffusion model is more efficient and more adequate for spot matching than the Gaussian model and the diffusion model.

• Journal of agricultural medicine and community health
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• v.34 no.1
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• pp.24-33
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• 2009

Objectives: In September 2008, an outbreak of diarrhea occurred among students attending Y school in Yeongcheon-si. Shigella sonnei was cultured from some of the rectal swabs. An epidemiological investigation was carried out to determine the source of the infection and the mode of transmission of the shigellosis outbreak. Methods: The index case lived in the D rehabilitation facility in Gyeongsan-si and an additional epidemiological investigation was carried out there. The cases could not be questioned due to their mental handicaps. The teachers were interviewed instead. A patient case was defined as a resident with diarrhea more than one time a day from September 18 to September 26, 2008 or a resident with confirmed Shigella sonnei at the Y school or the D rehabilitation facility. Results: The attack rate was 1.2% (8 persons) among 659 persons in the Y school and D rehabilitation facility. Five persons were microbiologically confirmed to have the infection and three persons were diagnosed on the basis of symptoms. Shigella sonnei was cultured from five of the 659 rectal swabs. However, 80 environmental specimens including drinking water, preserved foods, and cooking utensils were negative. All eight patients were Y school students and had been living in group boarding and lodging. Six of them lived in the D rehabilitation facility and two lived in the dormitory at the Y school. Five cases showed pulsed-field gel electrophoresis patterns that were identical for Shigella sonnei. Conclusions: The results of this study showed that the infection source of the shigellosis outbreak, in the two places, were identical. It is likely that the infections initially spread from a teacher or volunteer and then among the students.

• Applied Biological Chemistry
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• v.36 no.2
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• pp.105-110
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• 1993

Inulin fructotransferase from Enterobacter sp. S45 was purified with DEAE-cellulose column chromatography and fast protein liquid chromatography. The purified enzyme gave a single band on polyacrylamide gel electrophoresis. The molecular weight was estimated to be 42,800 by SDS-polyacrylamide gel electrophoresis. The optimal pH and temperature for the enzyme reaction were pH 5.5 and $55^{\circ}C$, respectively. $Mg^{2+}$ activated the enzyme activity, but $Fe^{3+}$, $Cu^{2+}$, $Hg^{2+}$ significantly inhibited. After exhaustive digestion of inulin by the enzyme, DFA III, sucrose, 1-kestose and nystose were produced. Sucrose, 1-kestose, raffinose and melezitose can't be used as substrates by the enzyme, but nystose and 1-F-fructofuranosyl nystose were hydrolysed. The Km and Vmax for inulin of the enzyme were 1.4 mM and $0.196\;{\mu}mole/min$, respectively.

• Korean Journal of Organic Agriculture
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• v.19 no.3
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• pp.417-425
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• 2011

This study was conducted to investigate the changes in physicochemical and microbiological properties during fermenting process of farm-made organic liquid fertilizer made of the mixture of organic materials such as blood meal and molasse during fermenting process. The pH level of organic liquid fertilizer during the ermentation decreased from 7.2 to 4.3. The EC of organic liquid fertilizer was increased from 13.9 dS/m to 99.3 dS/m during the fermentation. The total population of aerobic bacteria decreased from $8.2{\times}10^5$ cfu/ml to $3{\times}10^4$ cfu/ml, but Bacillus spp. increased from $2.1{\times}10^2$ cfu/ml to $4.2{\times}10^3$ cfu/ml during the fermentation. Bacterial isolates were obtained from organic liquid fertilizers and identified by fatty acid-base typing. The Genus Bacillus was dominant as fermenting proceeded. The denaturing gradient gel electrophoresis (DGGE) profile showed changes of bacterial communities in organic liquid fertilizers.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.477-486
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• 1991

To elucidate enzymatic properties of a-glactosidase (EC 3.2.1.22) from Asp. niger, a-galactosidase from wheat bran culture was purified by ammonium sulfate fractionation, ion exchange chromatography and gel filtration. And then its enzymatic propeties were investigated. The highest level of $\alpha$-galactosidase activity was obtained when Asp. niger was grown on wheat bran medium at $30^{\circ}C$ for 96 hours. The $\alpha$-galactosidase was purified by 23.7 fold by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Celluose and Sephadex A-50, and gel filtration on Sephadex G-150 and its specific activity was 1,229 Unitslmg protein and the yield was 14% of the total activity of wheat bran culture. The purified $\alpha$-galactosidase was found to be homogeneous by polyacrylamide gel electrophoresis and HPLC. The $\alpha$-galactosidase was a tetrameric glycoprotein which consisted of identical subunits with molelcular weight of 28,000 each by SDS-PAGE and isoelectric point was determined analytical isoelectric focusing to be pH 4.6. The optimal temperature and pH for the $\alpha$-galactosidase activity were $40^{\circ}C$ and pH 6.5, respectively, and 54% of its activity was lost by heating at $60^{\circ}C$ for 10 mins, It was appeared to have higher affinty to raffinose than to stachyose. The K, value and activation energy of $\alpha$-galactosidase were 5.0 mM and 8.515 Kcal per mole for p-nitrophenyl- $\alpha$--D-galactopyranoside, respectively.