• Applied Biological Chemistry
• /
• v.15 no.1
• /
• pp.27-33
• /
• 1972

The alkaline protease was isolated from the culture material of monascus sp. on wheat bran culture. The crude purification of this enzyme was extracted with distilled water and precipitated with ammonium sulfate of 0.5 saturation. And, the activity of this enzyme was determind very strongly by folin's colorimetric method. The optimal pH of this enzyme was ranging from pH 10 to 13 and the optimal temperature was $50^{\circ}C$. The pH stability was ranging from pH 5 to 12 and the enzyme activity was not inactivated by heat treatment in lower temperature than $40^{\circ}C$. The enzyme was protected from heat denature by the treatment of $Pb^#$, $Ba^#$, $Co^#$, $Zn^#$, and $Cu^#$, but was inactivated with $Hg^#$, $Fe^#$ strongly. Moreover, one of these metal ions, the cupper ion, has a strong protective activity on enzyme heat denature. And, it was not effected by treatment of EDTA.

• BMB Reports
• /
• v.45 no.9
• /
• pp.538-543
• /
• 2012

We evaluated the expression of the costimulatory molecules CD80 and CD83 and major histocompatibility (MHC) class II induced by 2,4-dinitrofluorobenzene (DNFB) in the RAW 264.7 macrophage cell line. In contrast to the previously reported effect of DNFB on dendritic cells, CD86 expression did not change. Furthermore, we observed that the CD83 expression level transiently increased and then decreased. Induction of CD80 and MHC class II molecule expression and a decrease in CD83 expression by DNFB in vitro were also confirmed in splenocytes of BALB/c and NC/Nga mice. However, DNFB did not influence CD83 expression in peritoneal $CD11b^+$ cells from BALB/c or NC/Nga mice. Detailed in vivo experiments and further studies on the possible contribution of $CD11b^+$ cells to induce atopic dermatitis (AD) would be helpful to attain a better understanding of AD pathogenesis.

• BMB Reports
• /
• v.43 no.8
• /
• pp.561-566
• /
• 2010

Though protein transduction domains (PTDs) are well known for the delivery of exogenous therapeutic proteins into living cells, the overall low efficiency of transduction is a serious obstacle. We investigated the effect of bog blueberry anthocyanins (BBA) on protein transduction efficiency and found that BBA enhanced the transduction efficiencies of Tat-SOD fusion protein into HeLa cells and mice skin. The enzymatic activities in the cells and skin tissue in the presence of BBA were markedly increased compared to controls. Further, BBA did not demonstrate any cell toxicity at various concentrations. Although the mechanism is not fully understood, we suggest that BBA might alter the conformation of the membrane, which would indicate that BBA can be used as a protein transduction enhancer for the efficient delivery of therapeutic proteins for a variety of disorders.

• BMB Reports
• /
• v.43 no.4
• /
• pp.250-256
• /
• 2010

Natural phosphodiester bond CpG-DNA that contains immunomodulatory CpG motifs (PO-DNA) upregulates the expression of proinflammatory cytokines and induces an Ag-driven Th1 response in a CG sequence-dependent manner in mice. In humans, only phosphorothioate backbone-modified CpG-DNA (PS-DNA) and not PO-DNA has immunomodulatory activity. In this study, we found that liposome-encapsulated PO-DNA upregulated the expression of human $\beta$-defensin-2 (hBD-2) and major histocompatibility class II molecules (HLA-DRA) in a CG sequence-dependent and liposome- dependent manner in human B cells. Of the three different liposomes, DOTAP has the unique ability to enhance the immunomodulatory activity of PO-DNA. In contrast, HLA-DRA and hBD-2 promoter activation can be induced by liposome-encapsulated PS-DNA in a CG sequence-independent manner, depending on the CpG-DNA species. Our observations demonstrate that, when encapsulated with a proper liposome in the immune system, natural PO-DNA has the potential to be a useful therapy for the regulation of the innate immune response.

• ALGAE
• /
• v.23 no.1
• /
• pp.15-29
• /
• 2008

Many species in Gymnodiniales, which are unarmored dinoflagellates, are responsible for marine algal blooms and some of them have potent toxin in the cell. Their taxonomy has so far been well-defined, and several genera (e.g. Akashiwo, Gymnodinium, Karenia) have recently been re-described. In Korea, few works have been carried out on their taxonomical and molecular studies. This study focused on comparison of both morphological and molecular characteristics of five unarmored dinoflagellates on Korean coastal water: Akashiwo sanguinea, Cochlodinium polykrikoides, Gymnodinium catenatum, Gymnodinium impudicum and Karenia aureolum (=K. mikimotoi). Morphological characteristics observed here was in good accordance with the original descriptions of individual species. In addition, none of difference was found in morphological comparisons between the Korean and foreign strains. Furthermore, molecular analysis showed that the SSU rDNA sequences were generally identical according to each species. In some distinct features, A. sanguinea, which has generally the same morphological features, were divided into two groups: one was Korean isolates including European isolates, the other was American isolates. In the two groups, the nucleus was positioned differently: middle of the cells in the Korean isolates (GnSg02, GnSg03), near the epicone in American isolates (CCMP1593, CCMP1837). In addition, this was strongly supported by phylogenetic analysis, inferred from the SSU rDNA sequences. K. aureolum (GrAr01) was corresponded to European G. aureolum (=K. mikimotoi) in shape and position of nucleus, chloroplast, however, which is similar to K. digitata in view of having a finger-like sulcus. This was in good agreement with phylogenetic study of these species. G. catenatum have identical morphology except the ridge location, and their genotype of SSU rDNA was also identical to GenBank data of the same species. From this study, we found that the five Korean unarmored dinoflagellates are identical morphological characteristics and genotype to each species of foreign isolates.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.6
• /
• pp.797-804
• /
• 2000

The human immunodeficiency virus type 1 (HIV-1) Tat is one of the viral gene products essential for HIV replication. The exogenous Tat protein is transduced through the plasma membrane and then accumulated in a cell. The basic domain of the Tat protein, which is rich in arginine and lysine residues and called the protein transduction domain (PTD), has been identified to be responsible for this transduction activity. To better understand the nature of the transduction mediated by this highly basic domain of HIV-1 Tat, the Green Fluorescent Protein (GFP) was expressed and purified as a fusion protein with a peptide derived from the HIV-1 Tat basic domain in Escherichia coli. The transduction of Tat-GFP into mammalian cells was then determined by a Western blot analysis and fluorescence microscopy. The cells treated with Tat-GFP exhibited dose- and time-dependent increases in their intracellular level of the protein. the effective transduction of denatured Tat-GFP into both the nucleus and the cytoplasm of mammalian cells was also demonstrated, thereby indicating that the unfolding of the transduced protein is required for efficient transduction. Accordingly, the availability of recombinant Tat-GFP can facilitate the simple and specific identification of the protein transduction mediated by the HIV-1 Tat basic domain in living cells either by fluorescence microscopy or by a fluorescence-activated cell sorter analysis.

• ALGAE
• /
• v.19 no.2
• /
• pp.107-114
• /
• 2004

Patterns of epilithic algal colonization on artificial substrata (unglazed ceramic tiles) were investigated from 23rd April to 3rd July 1999 at weekly intervals over a 10 weeks period outside and inside the mesocosm in Togyo reservoir within the Civilian Passage Restriction Line near Demilitarized Zone (DMZ) in Korea. The highest standing crops of epilithic algae was 1,798$\cdot$10³ cells$\cdot$$cm^{-2}$ outside the mesocosm on 26th June and also inside the mesocosm those was 2,391$\cdot$10³ cells$\cdot$$cm^{-2}$ on 26th June, 9 weeks after the experiment began. The dominants outside the mesocosm were Achnanthes minutissima, Navicula bicephala, Oscillatoria angusta, Synedra delicastissima, S. tenuissima, S. ulna v. danica and Tabellaria flocculosa, and those inside the mesocosm were Achnanthes minitissima, Coenochloris polycocca, Fragilaria crotonenesis, Peridinium cinctum, Synedra delicatissima, Tabellaria flocculosa and Ulothrix subtilissima. Diatoms were most abundant and Achnanthes minutissima was the most important species colonizing on the tiles. Chlorophyll-a content was highest value of 5.4 mg$\cdot$$m^{-2}$ on 19th June after 8 weeks growth outside the mesocosm and was 24.4 mg$\cdot$$m^{-2}$ on 26th June, 9 weeks after the experiment began on tiles inside the mesocosm. It was also shown that unglazed ceramic tiles were a more suitable substratum for colonization than the glass slides. Consequently the substratum selection plays an important role in the colonization by the epilithic algal community.

• Korean Journal of Microbiology
• /
• v.47 no.3
• /
• pp.268-274
• /
• 2011

Anabaena (Cyanobacteria, Nostocales) are important for water quality controls, because they are often responsible for freshwater green tides; moreover, some species are reported to produce hepatotoxin. In this study, we sequenced RNA polymerase beta subunit (rpoB) gene of Anabaena, and evaluated their sequences for the potential use of a molecular taxonomic marker in this taxon. Anabaena rpoB showed low DNA similarity and high genetic divergences when compared those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.01). Parsimony analyses showed the rpoB gene evolves 4.8-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each Anabaena strain more clearly compared with a 16S rRNA tree. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the species discrimination of Anabaena.

• BMB Reports
• /
• v.28 no.2
• /
• pp.112-117
• /
• 1995

Succinic semialdehyde reductase was inactivated by o-phthalaldehyde. The inactivation followed pseudo-first order kinetics, and the second-order rate constant for the inactivation process was 28 $M^{-1}s^{-1}$ at pH 7.4 and $25^{\circ}C$. The absorption spectrum ($\lambda_{max}$ 337 nm) and fluorescence excitation ($\lambda_{max}$ 340 nm) and fluorescence emission spectra ($\lambda_{max}$ 409 nm) were consistent with the formation of an isoindole derivative in the catalytic site between a cysteine and a lysine residue approximately about 3 $\AA$ apart. The substrate, succinic semialdehyde, did not protect enzymatic activity against inactivation, whereas the coenzyme NADPH protected against o-phthaladehyde induced inactivation of the enzyme. About 1 isoindole group per mol of the enzyme was formed following complete loss of enzymatic activity. These results suggest that the amino acid residues of the enzyme participating in a reaction with o-phthalaldehyde are cysteinyl and lysyl residues at or near the NADPH binding site.

• Microbiology and Biotechnology Letters
• /
• v.7 no.3
• /
• pp.149-155
• /
• 1979

Streptomyces sp. 115-5 was selected as the most active microorganism of about 200 strains for the production of chitinase. The enzyme was purified by (NH$_4$)$_2$SO$_4$ treatment, 1st-Sephadex G-100, DEAE-Cellulose, 2nd-Sephadex G-100 column chromatography, and evidence for homogenity was obtained from CM-Sephadex C-50 column chromatography and polyacylamide gel electrophoresis. The purified enzyme hydrolyzed chitin (N-acetyl glucosamine polymer) and chitosan (glucosamine polymer) but not cellulose. And with chitin as the substrate, a Km value of 3.6 mg of chitin per ml and a Vmax of 100 $\mu$mo1e fer hr were found. The activation of the chitinase was 3.66 kcal per mole. The molecular weight of the enzyme was esti-mated about 56,000 daltons by Sephadex G-100 chromatography and isoelectric point as pH 3.0.