• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.4
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• pp.557-563
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• 2014

This study was conducted to evaluate the effects of physical treatments on the antigenicity of gliadin in strong wheat flour. Strong wheat flour was treated with an autoclave (5, 10, 30, 50 min), a microwave (1, 5, 10 min), or both (10, 30, 50 min/ 5, 10 min), followed by SDS-PAGE, immunoblotting, and Ci-ELISA using anti-gliadin IgG. The results indicated that the binding ability of IgG to gliadin in strong wheat flour slightly decreased after autoclaving or autoclaving/microwaving. In particular, the binding ability was reduced to about 87% after autoclaving for 50 min and to 89% after autoclaving/microwaving (50/5 min). In addition, gliadin bands in the 50 min autoclaved group disappeared in both SDS-PAGE and immunoblotting. On the other hand, the antigenicity of gliadin was unaffected by microwaving alone. In conclusion, the results of this study suggest that autoclaving may reduce the antigenicity of gliadin in strong wheat flour.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.465-471
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• 1997

The cyclodextrin glycosyltransferase (CGTase, EC 3.2.1.19) from Bacillus brevis CD162 was purified by precipitating with ammonium sulfate, DEAE-Sepharose CL-6B column chromatography and Sephadex G-150 column chromatography. The molecular mass and pI of the purified enzyme were estimated to be 74,000 and 6.3 by SDS-PAGE and isoelectric focusing, respectively. The purified enzyme was clearly identified as the CGTase by zymogram after SDS-PAGE. The optimum pH and temperature for the enzyme activity were 8.0 and $55^{\circ}C$, respectively. The enzyme was stable at the range of pH $5.5{\sim}9.0$, and up to $50^{\circ}C$. The amino acid sequence from the $NH_2-terminal$ of the purified CGTase was Ser-Val-Thr-Asn-Lys-Val-Asn-Tyr-Ser-Lys-Asp-Val-Ile-Tyr-Gln. The yields of the products from starch as the substrate were 1.3% for ${\alpha}-$, 33.9% for ${\beta}-$, and 9.7% for ${\gamma}-cyclodextrin$.

• Biomolecules & Therapeutics
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• v.6 no.1
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• pp.89-94
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• 1998

CJ50001 is a recombinant human granulocyte colony-stimulating facto, (rHuG-CSF) that stimulates the formation of neutrophils from bone marrow stem cells. It was produced in E. colt and purified through refolding and several processes. We produced CS970125(300) using purified C150001 and additives in order to test the stability of CJ50001. When CS970125(300) was stored at 50'S for more than 1 week, high molecular weight proteins were formed and those proteins were detected by non-reducing SDS-PAGE, gel filtration HPLC, and Western blot. Those proteins showed single band at the same position of CJ50001 in reducing SDS-PAGE. These data indicated that those high molecular weight proteins were the multimers of C150001. In biological assays, iu viro and in viro, the multimers did not have biological activity and inhibitory action to that of CJ 50001. The mutimers did not induce toxicity in mice and rats in acute toxicity test. These results suggest that if Cs970125(300) containing CJ50001 is stored at 5$0^{\circ}C$, CJ50001 will be the multimers that do not have biological activity and inhibitory effect to CJ50001 and do not induce acute toxicity.

• The Korean Journal of Food And Nutrition
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• v.6 no.4
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• pp.307-313
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• 1993

Bacillus cereus $\beta$-amylase was purified by Sephadex G-100 gel filtration, CM Sephadex C-50 ion exchange chromatography and CM Sephadex C-50 ion exchange rechromatography The purified enzyme showed 871 unit/mg of specific activity. The purified enzyme was identified as homogenious by disc PAGE, SDS-PAGE and analysis of reaction product. The purified enzyme showed optimum pH 7.0. optimum temperature 5$0^{\circ}C$, and was stable at 0~5$0^{\circ}C$ and at pH range of 6~10.

• Parasites, Hosts and Diseases
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• v.38 no.2
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• pp.65-74
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• 2000

Toxoplasma-killing activities of mouse peritoneal macrophages activated by the extracts of Tetrahymena pyriformis (Korean and Chinese strains) were evaluated, and the active protein fractions from both strains were partially characterized by a method including chromatographies and SDS-PAGE. The first peak in Korean strain and the second peak in Chinese strain of T. pyriformis obtained by DEAE-Sephadex A-50 chromatography were most effective in the activation of macrophages to kill Toxoplasma gondii tachyzoites in vitro. Subsequent fractionations of obtained peak fractions were performed on a Sephadex G-200 gel. The first peaks fractionated from both strains of T. pyrtyomis had the highest toxoplasmacidal activities, and when subjected to the SDS-PAGE, one prominent band was visualized for each of the strains showing the same molecular weight of ca. 52.6 kDa. This active protein is suggested to be related to non-specific activation of mouse peritoneal macrophages.

• 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.

• Animal cells and systems
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• v.5 no.3
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• pp.199-203
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• 2001

Sperminogen was purified from the acid extracts of boar spermatozoa and partial peptide sequence of the 34-36 kd sperminogen was determined. Acid extracts of boar spermatozoa was gel-filtered through Sephadex G-75, and the 34-36 kd sperminogen was purified by preparative SDS-PAGE. The sperminogen bands were sliced out, and 34-36 kd sperminogen were eluted from the gel fragments and was subjected to peptide sequencing. Since the amino termini were blocked for Edman degradation method, internal amino acid sequences of the eluted 34-36 kd sperminogen were obtained from CNBr-digested peptides of sperminogen. Among several bands resolved on tricine SDS-PAGE, 14, 22 and 26 kd peptides were subjected to peptide sequencing. The ana1yzed amino acid sequences of the 26 and 22 kd peptides showed high homologies with that of the zona pellucida binding protein, Sp38, and the analyzed amino acid sequence of the 14 kd peptide showed neither sequence homology nor similarity with any known proteins.

• BMB Reports
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• v.33 no.6
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• pp.510-513
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• 2000

Boar sperminogen was purified from the acid extracts of the washed epididymal spermatozoa by gel filtration through a Sephadex G-100 column, followed by preparative SDS-PAGE. The 32 kDa sperminogen band was sliced out from the preparative SDS-PAGE and 32 kDa sperminogen was eluted from the gel matrix. The purified 32 kDa sperminogen was subjected to amino acid composition analysis. The amino acid composition of the 32 kDa boar sperminogen showed significant differences from that of either boar proacrosin or ${\beta}-acrosin$, which signifies that 32 kDa sperminogen might not be a breakdown product of proacrosin-acrosin system and that the 32 kDa sperminogen is a different protein from proacrosin-acrosin system.

• Proceedings of the PSK Conference
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• 2000.10a
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• pp.195.1-195.1
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• 2000

• Journal of Microbiology
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• v.35 no.4
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• pp.323-326
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• 1997

Toxin protein from Ustilago maydis virus SH14 isolated in Korea was purified using ethanol precipitation, cation exchange, gel filtration and anion exchange chromatography. The molecular weight of the purified protein was estimated to be 8.3 kDa by SDS-PAGE analysis. The Nterminal sequence of the protein is L-G-I-N-C(K)-R-G-S-S-Q--C(K)-G-L-S-G which is highly homologous with that of P4 toxin, but the amino acid composition and electrophoretic mobility in a native PAGE of the toxin protein were totally different from those of P4 toxin respectively. The SH14 toxin was shown to have immunological cross-reactivity about 50% with P4 toxin when examined by Western hybridization.