• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.612-619
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• 2000

To investigate the biochemical properties of V. mimicus metalloprotease, whose gene was isolated previously from Vibrio mimicus ATCC33653, overexpression and purification were attempted. The 1.9 kb of open reading frame was amplified by PCR from pVMC193 plasmid which ligated the VMC gene with pUC19 and introduced into Escherichia coli BL21 (DE3) using the overexpression vector, pET22b (+). The overexpressed metalloprotease (VMC) was purified with Ni-NTA column chromatography and characterized with various protease inhibitors, pHs, temperatures, and substrates. The purified VMC showed the proteolytic activity against gelatin, soluble and insoluble collagens, and synthetic peptides. Unlike the observations made with all metalloproteases originated from other Vibrio sp., the VMC did not hydrolyze the casein. The proteolytic activity was critically decreased when the VMC was treated with metal chelating reagents, such as EDTA, 2,2-bipyridine, and 1, 10-phenanthroline. In particular, the 71 kDa VMC exhibited the hemagglutinating activity against human erythrocyte. As the purified VMC was treated with $CuCl_2$ and $NiCl_2$ for the chemical modification of metal binding, the proteolytic activity and hemagglutinating activity were profoundly influenced. The multialignment analysis made on the reported Vibrio metalloproteases showed the difference of amino acid sequence similarity between the two distinctive classes of Vibrio metalloproteases.

• Microbiology and Biotechnology Letters
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• v.44 no.1
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• pp.55-61
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• 2016

Sepiapterin, a precursor for tetrahydrobiopterin, is produced in higher mammals using guanosine triphosphate (GTP) as a biosynthetic intermediate. Four genes involved in GTP biosynthesis, namely those of guanosine monophosphate kinase (gmk), nucleoside diphosphate kinase (ndk), guanosine phosphate synthetase (guaA), and inosine-5'-monophosphate dehydrogenase (guaB), were expressed in sepiapterin-producing recombinant Escherichia coli BL21(DE3) to increase intracellular GTP concentration and to improve sepiapterin production concomitantly. Coexpression of gmk, ndk, guaA, and guaB, doubled the intracellular GTP concentration and increased the maximum sepiapterin concentration up to $126.1{\pm}19.3mg/l$ (an increase of 43% compared with control cells) in batch-cultivated recombinant E. coli.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1557-1564
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• 2009

Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.128-135
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• 2009

Acinetohacter baumannii BD5 was isolated from waters of Baek-du mountain, and the lipase gene was cloned using a PCR technique. The deduced amino acid sequence of the lipase and lipase chaperone were found to encode proteins of 325 aa and 344 aa with a molecular mass of 35 kDa and 37 kDa, respectively. The lipase gene was cloned and expressed in Escherichia coli BL21(trxB) as an inclusion body, which was subsequently solubilized by urea, and then purified using Ni-affinity chromatography. After being purified, the lipase was refolded by incubation at $4^{\circ}C$ in the presence of a 1:10 molar ratio of lipase:chaperone. The maximal activity of the refolded lipase was observed at a temperature of $35^{\circ}C$ and pH 8.3 when p-NP caprate(C10) was used as a substrate; however, 28% of the activity observed at $35^{\circ}C$ was still remaining at $0^{\circ}C$. The stability of the purified enzyme at low temperatures indicates that it is a cold-adapted enzyme. The refolded lipase was activated by $Ca^{2+},\;Mg^{2+},\;and\;Mn^{2+}$, whereas $Zn^{2+}\;and\;Cu^{2+}$ inhibited it. Additionally, 0.1% Tween 20 increased the lipase activity by 33%, but SDS and Triton X-100 inhibited the lipase activity by 40% and 70%, respectively.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.873-880
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• 2009

A novel xylanase gene, Kxyn, was cloned from Kocuria sp. Mn22, a bacteria isolated from the deep sea of the east Pacific. Kxyn consists of 1,170 bp and encodes a protein of 390 amino acids that shows the highest identity (63%) with a xylanase from Thermohifida fusca YX. The mature protein with a molecular mass of approximately 40 kDa was expressed in Escherichia coli BL21 (DE3). The recombinant Kxyn displayed its maximum activity at $55^{\circ}C$ and at pH 8.5. The $K_m,\;V_{max}$, and $k_{cat}$ values of Kxyn for birchwood xylan were 5.4 mg/ml, $272{\mu}mol/min{\cdot}mg$, and 185.1/s, respectively. Kxyn hydrolyzed birchwood xylan to produce xylobiose and xylotriose as the predominant products. The activity of Kxyn was not affected by $Ca^{2+},\;Mg^{2+},\;Na^+,\;K^+$, ${\beta}$-mercaptoethanol, DTT, or SDS, but was strongly inhibited by $Hg^{2+},\;Cu^{2+},Zn^{2+}$, and $Pb^{2+}$. It was stable over a wide pH range, retaining more than 80% activity after overnight incubation at pH 7.5-12. Kxyn is a cellulase-free xylanase. Therefore, these properties make it a candidate for various industrial applications.

• Nutrition Research and Practice
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• v.5 no.1
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• pp.11-19
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• 2011

Eotaxin is an important inflammatory chemokine in eosinophil chemotaxis and activation and, thus, is implicated in asthma. Recently, obesity was associated with an increased prevalence of asthma, but the relationship between obesity and eotaxin expression has only been partially understood in obese mice and human studies. Therefore, we studied the expression patterns of eotaxin in 3T3-L1 preadipocytes/adipocytes to determine whether eotaxin levels are influenced by body weight gain and/or reduction in diet-induced obese mice. First, we investigated eotaxin expression during differentiation in 3T3-L1 adipocytes. Then, we treated 3T3-L1 preadipoeytes/adipoeytes with tumor necrosis factor-alpha (TNF-${\alpha}$), eotaxin, interleukin (IL)-4, IL-5, or leptin. To examine the effects of weight loss in high-fat diet induced obese mice, we fed C57BL/6 mice a high-fat diet or a normal diet for 26 weeks. Then, half of the high-fat diet group were fed a normal diet until 30 weeks to reduce weight. Epididymal adipose tissue, visceral adipose tissue, serum, and bronchoalveolar fluid of mice were examined for eotaxin expression. The results showed that eotaxin expression levels increased with adipocyte differentiation and that more eotaxin was expressed when the cells were stimulated with TNF-${\alpha}$, eotaxin, IL-4, IL-5, or leptin. An in vivo study showed that eotaxin levels were reduced in visceral adipose tissues when high-fat diet fed mice underwent weight loss. Taken together, these results indicate a close relationship between eotaxin expression and obesity as well as weight loss, thus, they indirectly show a relation to asthma.

• The Research Journal of the Costume Culture
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• v.26 no.4
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• pp.613-631
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• 2018

The purpose of this study is to collect and analyze costumes presented in international dance sports competitions, and summarize the features of Latin American dance costumes' design. As for research methods, standards of Latin American dance costume design were analyzed via a literature review on dance sports. The scope of the study extended for six years from 2010 to 2015 to include the, top three UK Latin American dance competitions. The results are as follows. First, the silhouette analysis determined that the X silhouette to the lead with, -145 costumes (78%), followed by the H silhouette at 25 (13%), and other at 16 (9%). Amongst those there were 174 one-piece dresses (94%). Furthermore, the analysis on colors of Latin American dance sports costumes revealed that, amongst the 186 costumes, 115 were without color (62%), Bl(black) is the most frequent with 37%, then Wh(white) with 21% and Gr(gray) with 4%. Costumes with colors, based on the six basic colors in the Munsell color system, are comprised most often of red with 12%, the followed by Y(yellow) at 10%, B(blue) at 8%, YR(yellow-red) at 4%, P(purple) at 2%, and G(green) at 2%. Thirdly, the cloth materials of Latin American dance costumes are recognized through image inspection. Among visually recognizable materials, beading materials are the most common with 104 costumes (60%). Shiny materials like mesh, chiffon, organza, lace and burn-out are in 36 costumes in total (19%). Other cloth materials included Luster materials and; non-sheen materials, which were in 46 costumes (25%).

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.19 no.3 s.31
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• pp.34-54
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• 2006

The effects of four extracts of medicinal herbs, Rubus coreanum, Acorus calamus, Morus alba and Rehmannia glutinosa on hair growth activity and acnes control were investigated. In the course of screening natural extracts for hair growth, we found that the extract of dried root of Rubus coreanum has the hair growth promoting effect. After topical application of these extracts to the back of C57BL/6 mice, the earlier conversion of telogen-to-anagen phase was induced. The growth of dermal papilla cells and mouse vibrissae hair follicle cultured in vitro, however, was not affected by treatment of these extracts. Furthermore these extracts do not possesspotent inhibitory effect on $5{\alpha}-reductase$ I and II activity and anti-bacterial effect on Escherichia coli , Propionibacterium acnes, Pityrosporum ovale, Staphylococcus aureus, Staphylococcus epidemidis, and Candida albicans. RT-PCR analysis showed that these extracts did notinduce mRNA levels of growth factors such as insulin-like growth factor-I, keratinocyte growth factor, hepatocyte growth factor and vascular endothelial growth factor in dermal papilla cells. These results suggest that Rubus coreanum has hair growth promoting effect. However, the effects of these materials on the hair growth promotion are not mediated through inhibition of $5{\alpha}-reductase$ I and II activity, stimulation of hair follicle cells and expression of growth factors in the dermal papilla cells.

• Textile Coloration and Finishing
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• v.11 no.2
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• pp.19-26
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• 1999

Strains degrading and decolorizing acid dyes, Nylosan red E-BL 150%. were isolated from natural system, was named as ARK3. The optimal culture conditions of temperature and pH were $35^\circ{C}$, 7.0, respectively. Growth rate of cells in conditions of aerobic shaking more than standing culture conspicuously increased, and optical density of those to strain ARK3 were found as 1.38 and 0.25 after 42 hrs. Decolorization efficiency in batch culture which used as immobilization media to natural zeolite was 15% after 6 hrs, while suspension culture was 5%, also its of immobilization and suspension culture were 90% and 85% after 48 hrs, respectively. Decolorization efficiency of air-lift bioreactor was more than 90% to a dilution rate of $0.038hr^{-1}$, but that was decreased as 70%, when the dilution rate was $0.05hr^{-1}$. Even though at maximum dilution rate of this study, there was not appeared "wash out" phenomienon of biomass. Decolorization efficiency was 97.7% at a dilution rate of $0.025hr^{-1}$, when influent dye concentration was $100mg/\ell$. But if influent dye concentration increased as $150mg/\ell$, even though MLVSS increased, that of treatment water decreased as 93%. Also, when influent dye concentration increased as $200mg/\ell$ and $300mg/\ell$, decolorization efficiencies of treatment water abruptly decreased as 85% and 63%, respectively. Decolorization efficiency was more than 92% to the limit volumetric loading rate of $3.75mg/\ell\cdot{hr}$hr, without regard to variation of influent dye concentration or hydraulic retention time. if volumetric loading rate was more than $3.80mg/\ell\cdot{hr}$, at same condition, decolorization efficiency was lower decrease of retention time than increase of influent dye concentration.entration.

• KSBB Journal
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• v.19 no.1
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• pp.27-32
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• 2004

2-Dimensional fluorescence sensor has a wide range of excitation and emission wavelengths, that some biogenic fluorphors in a biological process can be monitored simultaneously. The production processes of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL21 (DE3) pLysS harboring plasmid pFLS45 were on-line monitored by a 2-dimensional fluorescence sensor The characteristics of fluorescence spectrum was dependent upon physical and biological factors of a bioprocess such as culture pH, cell mass etc. Some off-line data were correlated to the fluorescence intensity well, which was monitored at some combination of excitation and emission wavelengths by the 2-dimensional fluorescence sensor.