• Parasites, Hosts and Diseases
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• v.33 no.2
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• pp.107-116
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• 1995

Two hybridoma cell lines against Cwptosporinium possum oocysts nFRl-CN911 were produced. The isotype of these 2 monoclonal antibodies (mAbs) was IgG2b (lE7.2) and and IgM (C6). Enzyme immuno-transfer blotting analysis showed that 157.2 reacted specifically to 36 kDa protein and C6 reacted to 67 and 70 kDa proteins. C. pcnlum was bound specifically to the surface region of oocysts by these mobs. No cross-reactivity was observed with tachyzoites of ToxopLosma gonnii and oocysts of Eimeria zuernii,5. bouis and E. canadensis of bovine origin. The indirect immunofluorescence assay (IIF) using mAb C6 was successful with counterstain. With the IIF using mob C6, oocysts appeared as 3 to $5{\mu}m$ spherical objects fluorescing bright apple green against a reddish dark background. The IIF using mAb C6 was agreed in specificity and sensitivity with those of a commercial diagnostic kit. These results demonstrated that the produced mAbs were specific to C. parvum and that the mAb C6 could be used for diagnosis of cryptosporidiosis.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.114-117
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• 2008

Chelating agents to sand lance fish sauce for the prevention of precipitate formation were applied. The precipitates consisted of crude protein (74.4%), ash (18.7%), and other components (6.9%). Sand lance sauce was mainly composed of glutamic acid (3.69 mg/g), alanine (2.96 mg/g), and lysine (2.64 mg/g). However, there was an increase in the amount of hydrophobic amino acids, phenylalanine, isoleucine, and leucine, in the precipitates. Sodium ions were not detected in the precipitate; rather, the main elements were Mg ($1.98{\times}10^4\;{\mu}g/g)$, K ($1.36{\times}10^4\;{\mu}g/g)$, and Ca ($6.66{\times}10^2\;{\mu}g/g)$. In HPLC analysis, fish sauce was composed of 2 main peaks with molecular weights of 85.5 and 528.4 Da, respectively. However, the precipitate contained one peak with a molecular weight of 1,513.5 Da. The addition of 0.2% malic acid and citric acid caused 55 and 70% prevention of the precipitate, respectively. Citric acid was the most effective chelating agent and efficiently prevented precipitation in the fish sauce.

• Journal of Pharmacopuncture
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• v.8 no.2
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• pp.23-28
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• 2005

Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down-regulated protein was heat shock 70kDa protein 5 and up-regulated proteins were chaperonin and 2-phospho -pyruvate-hydratase ${\alpha}-enolase$ by 1.5mg/ml of CTF-HAS. Discussion : Proteomic analysis approach were performed to screen the differential expression genes. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

• Reproductive and Developmental Biology
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• v.38 no.1
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• pp.53-62
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• 2014

The objective of this study was to investigate the relationship between fertility and protein pattern change using in vitro fertilization, analysis of sperm characteristics and two-dimensional gel electrophoresis in different pig types. In results, the viability and mitochondria integrity of sperm were higher significantly (p<0.05) but the portions of acrosome reaction was lower significantly (p<0.05) in Duroc and $F_1$ (potbellied ${\times}$ PWG miniature pig) than PWG miniature. On in vitro fertilization to investigate fertility, the fertility of $F_1$ semen war higher significantly (p<0.05) than in Duroc and PWG miniature pig. On the other hand, protein patterns showed similar function among the different boar semen. Especially, the heat shock 70 kDa 1-like and G patch domain-containing protein 4 were significantly (p<0.05) higher expressed in $F_1$ than in Duroc and PWG miniature pig. The proteins associated with mitochondria in Duroc were significantly (p<0.05) higher expressed than in $F_1$ and PWG miniature pig. The developmental rates to blastocyst stage of oocytes fertilized with sperm of $F_1$ pig were significantly (p<0.05) higher than in PWG miniature pig. However, phosphoglycerate kinase 2 and zinc finger protein 431 were significantly (p<0.05) higher expressed in PWG miniature pig than in $F_1$ and Duroc pigs. In conclusion, the results of the present study indicate that different proteins were expressed in different pig types, and were associated with a sperm functions and embryo development.

• Journal of the Korean Wood Science and Technology
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• v.35 no.6
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• pp.159-165
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• 2007

An extracellular xylanase from the brown-rot fungus Fomitopsis palustris grown on rice straw culture was purified to a single protein band. On SDS-PAGE, the molecular mass of purified enzyme was estimated to be about 43 kDa. The amino acid sequence of the proteolytic fragments showed high homology with fungal glycoside hydrolase family 10 xylanases. The $K_m$, $K_{cat}$ and $V_{max}$ for birch xylan were $31mg/m{\ell}$, $2.3{\times}10^4/min$ and 252.3 U/mg, respectively. The optimal activity of the purified xylanase from F palustris was observed at pH 4.0~5.0 and $70^{\circ}C$.

• Journal of Life Science
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• v.27 no.3
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• pp.318-324
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• 2017

This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

• Journal of Ginseng Research
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• v.23 no.4
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• pp.217-221
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• 1999

We have isolated an antithrombin active polysaccharide in red ginseng by procedures comprising three major steps involving alkaline extraction, anion exchange and gel permeation chromatography. Active polysaccharide behaved as a single band on cellulose acetate membrane electrophoresis. The average molecular mass was estimated to be about 177 kDa by gel filtration. This polysaccharide was found to be an acidic heteropolysaccharide that contains uronic acid moiety $(40.2\%)$, sulfate group $(9.2\%)$ and protein $(1.5\%)$ in addition to neutral sugar consisted of rhamnose, mannose, galactose, arabinose, glucose, fucose and xylose in a molar ratio of 1.00 : 0.88 : 0.86 : 0.78: 0.70 : 0.33 : 0.22. This polysaccharide inhibited blood coagulation via the intrinsic pathway like heparin in a dose-dependent manner. The clotting of fibrinogen by thrombin was also mitigated by the presence of this polysaccharide.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.115-122
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• 2000

A Bacillus thuringiensis designated NT0423, belonging to B. thuringiensis subsp. aizawai (H 7), was isolated from samples of dust and soil of sericultural farms. B. thuringiensis NT0423 having dualspecificity against Lepidoptera and Diptera produced bipyramidal inclusions consisting of two major polypeptides of approximately 130- and 70-kDa. Proteolytic processing by trypsin and gut juice of Bombyx mori yielded predominant proteins with molecular masses of about 66-kDa. The whole crystal protein of B. thuringiensis NT0423 immunologically was related to that of B. thuringiensis subsp. aizawai. PCR analysis showed that B. thuringiensis NT0423 has at least five crystal protein genes including cryIA(a), cryIA(b), cryIC, cryID and cryIIA, and southern blot was determined the location of each gene on intact and enzyme-digested plasmid DNA fragments. Except for cryIA(a) gene on the high molecular weight plasmid of 165-kb, all of four genes were located on the plasmid of 66-kb. The production of $\beta$-exotoxin from B. thuringiensis NT0423 was identified by the HPLC analysis. In addition, the $\beta$-exotoxin showed its ability to prevent pupation of treated larvae of house flies (Musca domestica) from developing into normal adults.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.445-448
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• 2002

The Pseudomonas fluorescens KCTC 1767, a selected and identified as potential candidate for stereo-specific resolution of rac-ketoprofen ethyl ester, was systematically investigated in order to induce the high level expression and detailed characterization of the expressing enzyme esterase. We cloned the esterase gene from chromosomal DNA of Pseudomonas fluorescens KCTC 1767 by PCR with two synthetic primers that desinged for simple purification. The recombinant esterase from Pseudomonas fluorescens KCTC 1767 exibited a high conversion rate and enantioselectivity to the (S)-ketoprofen ethyl ester as expected. The enzyme was easily purified to homogeniety by using a metal chelating affinity chromatography as a protein with poly histidine taq, and thus obtained 0.6 mg of protein from a 100 mL culture broth in a single step. The purified enzyme was steadily stable at the pH range from 7.0 to 10. The activity was also retained to be about 70% after the preincubation at $40^{\circ}C$ but over $50^{\circ}C$ lost the activity completely. The molecular mass of the esterase was estimated to be about 43 kDa on SDS-PAGE, and an identical result was also shown in gel filteration chromatography. The specific activity was calculated 27 mM/mg-protein/min by using the rac-ketoprofen ethly ester as a substrate.

• Journal of Microbiology and Biotechnology
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• v.20 no.10
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• pp.1415-1423
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• 2010

An extracellular xylanase was purified to homogeneity by sequential chromatography of Fomitopsis pinicola culture supernatants on a DEAE-Sepharose column, a gel filtration column, and then on a MonoQ column with fast protein liquid chromatography. The relative molecular mass of the F. pinicola xylanase was determined to be 58 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by size-exclusion chromatography, indicating that the enzyme is a monomer. The hydrolytic activity of the xylanase had a pH optimum of 4.5 and a temperature optimum of $70^{\circ}C$. The enzyme showed a $t_{1/2}$ value of 33 h at $70^{\circ}C$ and catalytic efficiency ($k_{cat}=77.4\;s^{-1}$, $k_{cat}/K_m$=22.7 mg/ml/s) for oatspelt xylan. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase (GH) family 10, indicating that the F. pinicola xylanase is a member of GH family 10.