• Microbiology and Biotechnology Letters
• /
• v.8 no.1
• /
• pp.27-32
• /
• 1980

We isolated a strain of Pseudomonas sp. from soil to utilize furfuryl alcohol as a carton source by enrichment culture. Alcohol dehydrogenase from this bacteria was purified 700-fold by Sephadex G-200 and affinity column chromatography to be homogeneous by electrophoresis and analytical centrifugation. This enzyme had a molecular weight of 120,000 and was composed of four subunits consisting of 266 amino acid residues. The optimal pH of the enzyme was pH 8.5 to 9, and the optimal temperature was, 45$^{\circ}C$. This enzyme was stable at 55$^{\circ}C$, but lost 80% of its activity in 10min at 6$0^{\circ}C$.

• Microbiology and Biotechnology Letters
• /
• v.17 no.4
• /
• pp.313-320
• /
• 1989

Rhodotorula glutinis K-24 was found to produce internal, cell wall bound, and external invertase. Internal invertase was purified by column chromatographies on DEAE-Sephadex A-50, Sp-sephadex C-50, gel filtration on Sephadex G-200 and isoelectric focusing. Cell wall bound invertase was partially purified by the following procedures; column chromatography on DEAE-Sephadex A-50 and gel filtration on Sephadex G-100. Optimum pH and temperature for enzymatic activities of internal and cell wall bound invertase were pH 3.0 and 6$0^{\circ}C$, respectively. Both enzymes were inhibited by HgC1$_2$, AgNO$_3$, MnSO$_4$, and sodium dodecylsulfate. The molecular weights of internal and cell wall bound invertases were estimated to be 310,000 and 61,000, respectively. Other physicochemical properties of the both enzymes were similar.

• Food Science and Preservation
• /
• v.12 no.5
• /
• pp.489-495
• /
• 2005

Polyphenol oxidase (PPO) from Burdock (Arctium lappa L.) was purified and characterized. Purification of polyphenol oxidase was achieved by ammonium sulfate precipitation, Phenyl-sepharose CL-4B hydrophobic chromatography and Sephadex G-100 gel filtration chromatography. The molecular mass of the purified PPO was estimated to be 30 kDa by SDS polyacrylamide gel electrophoresis. In a substrate specificity, maximum activity was achieved with chlorogenic acid, followed by catechol and catechin. Whereas, there was low activity with hydroquinic acid, resorcinol or tyrosine. The optimum pH and temperature for enzyme activity were 7.0 and 35$\circC$ with catechol, respectively. The enzyme was most stable at pH 7.0 while unstable at acidic and alkaline pH. The enzyme was stable when heated to 40$\circC$. But heating at 50$\circC$ for more than 30 min caused 50% loss of activity. Ascorbic acid, L-cystein and $Cu^{2+}$ inhibited the activity of pholyphenol oxidase.

• Analytical Science and Technology
• /
• v.17 no.1
• /
• pp.37-44
• /
• 2004

An alcohol oxidase from Hansenula polymorpha was strongly induced when cells were grown with 0.5% methanol supplementation as the carbon source. The induced Hansenula polymorpha alcohol oxidase was purified to electrophoretic homogeneity by using DEAE-Sephacel and Mono Q column chromatographys. The enzyme oxidized mainly primary aliphatic alcohols and exhibited high substrate specificity towards ethanol and methanol. The activity of the enzyme optimally proceeded at pH 8.5 and $50^{\circ}C$. The midpoint of the temperature-stability curve for the enzyme was approximately $52^{\circ}C$ and the enzyme was not completely inactivated even at $65^{\circ}C$ temperature. The enzyme showed resistance toward detergents and highly stable over 7 weeks of storage condition. This Hansenula polymorpha alcohol oxidase may be useful for the enzymatic determination of alcohol and for the industrial production of alcohols and aldehydes.

• Journal of Life Science
• /
• v.8 no.2
• /
• pp.119-125
• /
• 1998

Column-purified double-stranded RNA binding factor (RBF) protein was tested for its binding affinity for the different forms of nucleic acids structure such as single-stranded(ss) and double-stranded(ds)RNA and ss- and dsDNA. The RBF protein was incubated with each of these nucleic acid structures in separate reactions and its comparative binding affnity was visualized by SDS-polyacrylamide gel electrophoresis. The RBF protein bound to the dsRNA molecule to form a tight RNA:protein complex in agreement with previous studies, but not to the other nucleic acid molecules confirming its distinctive affinity for the dsRNA structure. In phosphorylation assay in vito, the purified RBF protein significantly inhibited the autophosphorylation of the PKR derived from not only human but mouse source in the presence of poly(I):poly(C). It is suggesting that PKR vs. RBF is similarly under a competitive interaction among different eukaryotic organisms during protein synthesis.

• The Korean Journal of Mycology
• /
• v.22 no.4
• /
• pp.298-308
• /
• 1994

The properties of polygalacturonase by Ganoderma lucidum in liquid culture were investigated. The enzyme was composed of an endo- and an exo-polygalacturonase. The endo- and exo-polygalacturonase were purified approximately 56 and 9.2-fold, respectively, through ammonium sulfate fractionation, gel filtration on Biogel P-100, anion exchange chromatography on DEAE-cellulose, gel chromatography on Sephadex G-150 and re-gel chromatography on Sephadex G-150. The endo- and exo-polygalacturonase had higher affinity for apple pectin than for citrus pectin or pectic acid. The Km values of the endo- and exo-polygalacturonase for apple pectin, determined on the Lineweaver-Burk plot, were 1.44 and 10.6 mg $ml^{-1}$ for apple pectin, respectively. Purified endo-polygalacturonase was found to be homogeneous electrophoretically and had a molecular weight of 54,000 estimated on SDS polyacrylamide gel. The optimal pH for the activity of the enzymes was 4.0. The endo- and exo-polygalacturonase were stable in the pH range of 4.0 to 6.0 and 3.5 to 5.5, respectively. The optimal temperatures of the endo- and exo-polygalacturonase were 40 and $60^{\circ}C$, respectively. The exo-polygalacturonase was more resistant to heat than the endo-polygalacturonase, requiring heating for 40 min at $80^{\circ}C$ for complete inactivation. The activity of the endo-polygalacturonase was increased by $Ca^{++}$ and $Mn^{++}\;ions$, while that of the exo-polygalacturonase was increased by $Ca^{++}\;ion$ only, and was not affected by $Mn^{++}\;ion$.

• Journal of Life Science
• /
• v.18 no.6
• /
• pp.770-775
• /
• 2008

Several studies reported quantitative trait loci (QTL) for meat quality on porcine chromosome 2. For application of the chromosomal information to pig industry through using DNA technology, single nucleotide polymorphism (SNP) markers are developed by comparative re-sequencing of polymerase chain reaction (PCR) products of 13 candidate genes. A total of 34 SNPs were identified in 11 PCR products, an average of one SNP in every 296 bp.PCR restriction fragment length polymorphism (RFLP) assays were developed for 11 SNPs and used to genotype four commercial pig populations in Korea. The SNP markers were used to map candidate genes in QTL and to clarify the relevance of SNP and quantitative traits.

• Microbiology and Biotechnology Letters
• /
• v.17 no.5
• /
• pp.494-498
• /
• 1989

Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

• Journal of the Korean Chemical Society
• /
• v.52 no.6
• /
• pp.630-636
• /
• 2008

• Microbiology and Biotechnology Letters
• /
• v.18 no.4
• /
• pp.360-367
• /
• 1990

The several glycoside hydrolysing enzymes related to rutin degradation are found to be rhamnosidase, glucosidase and rutinosidase. Rutinosidase was purified to electrophoretic homogeneity from cell extracts of rutin-degrading strain, MT-57, which was identified as a Arthrobacter sp. Its molecular weight was estimated to be 42, 000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 40, 000 by gel filtration. The optimum pH for enzyme was found to be 7.5, and relatively stable in alkaline solution. The optimum temperature for enzyme was $45^{\circ}C$, being stable up to $50^{\circ}C$ for 20 min. The Bm value of enzyme for rutin was 0.5 $\mu \textrm m$. The enzyme activity was increased by the chelating agent such as EDTA, $NaN_3$, and 8-hydroxyquinoline, was strongly inhibited by $CO_{2+}, Ni^{2+}$, and $Cu^{2+}$. The enzyme had high substrate specificity in the rutinoside.