• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.312-317
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• 1995

The polygalacturonase (PG) production in rotten apples by Botryosphaeria dothidea was purified by using gel filtration and ion exchange column chromatography, and the biochemical characters of PG were investigated. The purified PG appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with approximate molecular weight of 49 kilodalton (kDa). The molecular weight was equal to the native molecular weight estimated by gel filtration. The Km and Vmax values of PG were 0.51 mg/ml and 90.9 $\mu$M/min/ml, respectively. Optimum pH was 4.0~5.0, and the PG activity was stable from pH 5.0~10.0. Optimum temperature of the enzyme activity was 4$0^{\circ}C$. The PG activity was relatively stable at 2$0^{\circ}C$, but it was reduced 45% at 4$0^{\circ}C$ and completely inactivated at 8$0^{\circ}C$. The PG activity was considerably inhibited by Cu2+, Zn2+, SDS and EDTA, whereas it was not effected by Ca2+, K+, Mg2+ or Na+ ions.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.313.2-313.2
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• 2002

Environmentally benign protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, zincon (ZC) and a basic dye. ethyl violet (EV) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution produces bluish violet colored bands. It is a rapid procedure, involving only fixin and staining steps that are completed in 45 min. (omitted)

• Journal of Applied Biological Chemistry
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• v.55 no.4
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• pp.263-266
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• 2012

Isolation of iron and calcium-binding peptides derived from cottonseed meal protein (CMP) hydrolysates was investigated. The degree of hydrolysis of CMP by Flavourzyme was monitored using trinitrobenzenesulfonic acid method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Enzymatic hydrolysis of CMP for 12 h was sufficient for the preparation of CMP hydrolysates, and the hydrolysates were membrane-filtered under 3 kDa as a molecular weight. The filtered solution was fractionated using Q-Sepharose fast flow, Sephadex G-15, and reversed phase-high performance liquid chromatography for iron and calcium-binding peptides. As a result, F51 fraction was obtained as the best candidate for calcium and iron chelation, and the isolated iron and calcium-binding peptides can be used as functional food additives, similar to iron and calcium supplements.

• Parasites, Hosts and Diseases
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• v.30 no.4
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• pp.323-328
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• 1992

Analysis of siR isolates of Trichemenes veginalis was carried out with sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunoelectrotransfer blot (EITB). Trichloroacetic acid-treated antigens of the 6 isolates revealed 25 protein profiles ranging 12~170 kDa of molecular weight in SDS-PAGE. In EITB, the specific immunogenic bands were visualized at 51 kDa and 96 kDa when HY-1 antigen was probed with difFerent mice sera immunized with 6 isolates of T. vaginalis. The banding patterns with different sera showed isolate-to-isolate variability. In EITB, homologous antigen (HY-1) did not show any enhanced response in reacting to homologous antiserum(HY-1) when 6 isolates of T. vaginalis were probed with a single serum (HY-1). It is assumed that the different banding patterns of six isolates show isolate-to-isolate variability and immunogenic common bands in 41, 47, 74 and 9:k kDa on EITB may connote the important significance on immune response in T. vaginalis infection.

• Korean Journal of Animal Reproduction
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• v.16 no.1
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• pp.1-6
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• 1992

These experiments were undertaken as a basic study to understand the role of cumulus cell on in vitro maturation and fertilization process with identifying the cumulus cell-secreted proteins. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and fast protein liquid chromatography(FPLC), the proteins of cumulus cells were identified. The results obtained in these experiments were summarized as follows ; 1. When the proteins secreted from cultured cell for 30 hours were separated by SDS-PAGE, there were a major band (>94,000) and other minor 2 bands with molecular weight ranging 30,000∼67,000 dalton. 2. In addition, the fractionations of these proteins by FPLC were idirectly shown that three bands were hyaluronic acid, chondroitin sulfate, and heparin.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.119-126
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• 1986

A periplasmic endoprotease, named protease Pi, was purified to homogeneity from Escherkchia coli by conventional procedure with insulin as substrate. This enzyme degrades insulin and glucagon to trichloroacetic acid-soluble meterials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. Its molecular weight was 110, 000 when determined by gel filtration on Sephacryl S-300 and was 105, 000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, it appears to be single polypeptide. This snzyme is metalloprotease, since it is completely inhibited by o-phenanthroline and can be activated by addition of divalent metal cations, such as $Mg^{2+}\;and\;Co^{2+}$. It is destinct from protease Ci, a cytoplasmic insulin degrading enzyme, since protease Pi is localized to the periplasm. Since protease Pi selectively degrades GTP cyclohydrolase I, it appears to play a role in the regulation of pteridine biosynthesis.

• Preventive Nutrition and Food Science
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• v.15 no.1
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• pp.74-77
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• 2010

Antibiotic-resistant Staphylococcus aureus is beginning to pose a social issue. Thus, there is an urgent need for the development of effective anti-staphylococcal agents to eradicate antibiotic-resistant S. aureus in food systems and to treat the pathogen in clinical areas. To address this need, lactic acid bacteria (LAB) from kimchi were screened for the production of anti-staphylococcal bacteriocin. From this screening, a bacteriocin generated by the MK3 strain, which was identified by 16S rRNA gene sequence analysis as Enterococcus faecium, demonstrated antimicrobial activity against an S. aureus strain, and was designated enterocin MK3. Enterocin MK3 also demonstrated activity against other gram-positive bacteria, including several LAB and Listeria monocytogenes, but not gram-negative Escherichia coli. The molecular mass of enterocin MK3 was estimated as approximately 6.5 kDa on an SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) gel.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.217.3-218
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• 2003

A novel lectin has been purified from the fruiting bodies of the mushroom, Fomitella fraxinea, which belongs to bracket fungi by a combination of ion-exchange chromatography on DEAE-cellulose and gel filtration chromatography on Sephacryl S-200 HR. The lectin, designated as FFrL, was a homotetrametric protein with a molecular weight of 50 kDa as demonstrated by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and MALDI-TOF-MS(matrix assisted UV laser desorption/ionization time-of flight mass spectrometry). (omitted)

• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.386-390
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• 1997

A xylanase from the Bacillus sp. strain DSNC 101, isolated from soil, was purified to homogeneity by anion-exchange and hydrophobic interaction chromatography followed by gel filtration chromatography. The enzyme cleaved xylan, but not carboxymethyl cellulose, Avicel, soluble starch, and pNPX. The main product of oat spelts xylan hydrolysates was xylobiose. The xylanase had a molecular weight of 25 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Optimum temperature and pH for the xylanase activity were $50^{\circ}C$ and 6.0, respectively. $K_{m}\;and\;V_{max}$ of the enzyme for oat spelts xylan were 12.5 mg of xylan/ml and 869.5 unit/mg of protein, respectively. Xylanase was completely inhibited by Hg, Cu, and N-bromosuccinimide, but was stimulated by Ca, Co, and Mg.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.215-222
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• 2020

The marine microorganism PX-1, which can hydrolyze xylan, was isolated from coastal sea water of Jeju Island, Korea. Based on the 16S rRNA gene sequence and chemotaxonomy analysis, PX-1 was identified as a species of the genus Aestuariibacter and named Aestuariibacter sp PX-1. From the culture broth of PX-1, an extracellular xylanase was purified to homogeneity through ammonium sulfate precipitation and subsequent adsorption chromatography using insoluble xylan. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography estimated the molecular weight of the purified putative xylanase (XylA) as approximately 64 kDa. XylA showed xylanase activity toward beechwood xylan, with a maximum enzymatic activity at pH 6.0 and 45℃. Through thin-layer chromatographic analysis of the xylan hydrolysate produced by XylA, it was confirmed that XylA is an endo-type xylanase that decomposes xylan into xylose and xyloligosaccharides of various lengths. The K_m and V_max values of XylA for beechwood xylan were 27.78 mM and 78.13 μM/min, respectively.