• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.293-297
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• 2002

The cyclodextrin glucanotransferase (CGTase) gene from Bacillus stearothermophilus NO2 was expressed in Saccharomyces cerevisiae 2805 under the adhl promoter. The CGTase was purified from S. cerevisiae 2805/pVT-CGTS. The purified enzyme exhibited a optima of activity around pH 7.0 and $65^{\circ}C$. Thermal stability of the enzyme was increased fairly as compared with the CGTase of B. stearothermophilus NO2. The conversion yield of cyclodextrin (CD) and the production ratio of $\alpha$-, $\beta$,-, ${\gamma}$-CD from starch were showed similarly aspect to the CGTase of B. stearothermophilus NO2.

• Journal of the Korean Applied Science and Technology
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• v.36 no.3
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• pp.966-974
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• 2019

Arthrospira platensis has been reported to contain a variety of substances such as phycocyanin, ${\beta}$-carotene, vitamin E and other carotenoids. In this study, zebrafish were treated with indoor cultivation spirulina ethanol extracts(ICAE) to determine toxicity(coagulation rate, hatching rate, heart rate). We used the DCFH-DA staining method to detect the effect of reactive oxygen species(ROS) generation on lipopolysaccharide(LPS)-induced zebrafish embryos ROS various concentrations(0.01, 0.05, 0.1, 0.5mg/ml) of ICAE. Cell toxicity was measured by WST-1 assay on RAW 264.7 macrophage cell line. Also, measured the inhibitory effect of nitric oxide(NO) and prostaglandin $E_2$ ($PGE_2$) production in RAW264.7 macrophages induced by LPS at various concentrations of ICAE. The results of embryo coagulation rate, hatching rate, heart rate of zebrafish at various(0.01, 0.05, 0.1mg/ml) of ICAE was no toxicity. The ICAE treated group had an inhibitory effect on NO and $PGE_2$ production compared and decreased with concentration. The results of this study ethanol extract of Arthrospira platensis has an anti-inflammatory effect and suggest that is worthy of use cosmetics for skin protection.

• Parasites, Hosts and Diseases
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• v.48 no.2
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• pp.151-155
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• 2010

Allergen extracts from dust mites and cockroaches commonly found in Korean homes were used to evaluate their enzymatic activity as they are believed to influence allergenicity. Allergen extracts were prepared from 3 dust mite species (Dermatophagoides farinae, D. pteronyssinus, and Tyrophagus putrescentiae) and 3 cockroach species (Blattella germanica, Periplaneta americana, and P. fuliginosa) maintained in the Korea National Arthropods of Medical Importance Resource Bank. Proteins were extracted in PBS after homogenization using liquid nitrogen. The activities of various enzymes were investigated using the API Zym system. No significant difference in phosphatase, lipase, or glycosidase activity was observed among the 6 allergen extracts, but much difference was observed in protease activity. Protease activity was assessed in more detail by gelatin zymography and the EnzChek assay. Extract from T. putrescentiae showed the highest protease activity, followed by those of the cockroach extracts. Extracts from D. farinae and D. pteronyssinus showed only weak protease activity. Gelatinolytic activity was detected mainly in a 30-kDa protein in D. farinae, a 28-kDa protein in D. pteronyssinus, a > 26-kDa protein in T. putrescentiae, a > 20-kDa protein in B. germanica, and a > 23-kDa protein in P. americana and P. fuliginosa. The information on various enzymatic activities obtained in this study may be useful for future studies. In particular, the strong protease activity found in cockroach extracts could contribute to sensitization to cockroach allergens, which is known to be associated with the development of asthma.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.97-104
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• 2008

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin. PA/plasmin system playa role in mammalian fertilization and motility and acrosome reaction of sperm. The present study was undertaken to identify PAs in porcine gametes and investigate a possible role of plasminogen in in vitro fertilization in the pig. When boar spermatozoa were preincubated in a fertilization medium (mTBM) for 0, 2, 4 or 6 h, the activity of tPA-PAI ($110{\sim}117\;kDa$), tPA ($62{\sim}70\;kDa$), and uPA ($34{\sim}38\;kDa$) was observed in the sperm incubation medium and sperm sample. PA activities in the sperm incubation medium significantly (p<0.05) increased according to increasing incubation times, while PA activities in sperm significantly (p<0.05) decreased at the same times. In addition, the rate of acrosome reaction in spermatozoa increased by increasing culture times. When oocytes were separated from porcine cumulus-oocytes complexes at 0, 22 or 44 h of maturation culture, no PA activities were observed in cumulus free-oocyte just after aspiration from follicles. However, the activity of tPA-PAI ($108{\sim}113\;kDa$) and tPA ($75{\sim}83\;kDa$) was observed at 22 h of in vitro culture and significantly (p<0.05) increased as the duration of the culture increased. On the other hand, when porcine oocytes were activated by sperm penetration or calcium ionophore, plasminogen significantly (p<0.05) increased ZP dissolution time (sec) in activated oocytes by sperm penetration. These results suggest that supplementation of plasminogen to fertilization medium may playa positive role in the improvement of in vitro fertilization ability in the pig.

• Korean Journal of Weed Science
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• v.17 no.1
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• pp.44-51
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• 1997

Glyphosate had no effect on 5-enolpyruvylshikimate-3-phosphate synthase(EPSP-synthase) biosynthesis per se. But it inhibited clealy the activity of EPSP-synthase. EPSP-synthase seemed to be synthesized as a higher molecular weight(54 kDa) presusor protein and to be transported into plastid. The apparent molecular weight of mature EPSP-synthase in plastid is 45 kDa. Thus, the molecular size of transit peptide appeared to be about 9 kDa. The etiolation for 48 h after glyphosate application did not exhibit the inhibition of translocating level of EPSP-synthase across chloroplast envelope in actively growing meristematic leaves. But even when the plants were etiolated 2 hr after glyphosate treatment, a complete inhibition did not occur at least within 12 hr, i.e. 2 hr after beginning light period, suggesting that EPSP-synthase biosynthesis appeared to be not completely light dependent and the level of EPSP-synthase translocation to chloroplast could be controlled by an unknown regulatory mechanism of light dependent herbicidal effect of glyphosate.

• The Plant Pathology Journal
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• v.20 no.2
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• pp.158-163
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• 2004

Chlorella is a eukaryotic microalgae which shares metabolic pathways with higher plants. These charac-teristics make chlorella a potential candidate for eukaryotic overexpression systems. Recently, a foreign flounder growth hormone gene was stably introduced and expressed in transformed Chlorella ellipsoidea by using a modified plant transformation vector that contains cauliflower mosaic virus (CaMV) 35S pro-moter and the phleomycin resistant Sh ble gene as a selection marker. In this study, this same vector was modified by incorporating a promoter and a 3' UTR region of the 33kDa peptide gene from a chlorella virus that was isolated in our laboratory. The 33kDa gene promoter was used to replace the 35S promoter and the 3' UTR was introduced to separate the target gene and downstream Sh ble gene. Three different chlorella transformation vectors containing human erythropoietin (EPO) gene were constructed. The mp335EPO vector consists of a promoter from the 33kDa peptide gene, whereas the mp3353EPO vector contains the same promoter from the 33kDa peptide gene and its 3' UTR. The mp35S33pEPO vector contains the 35S promoter and the 3' UTR from the 33 kDa peptide gene. There was no significant difference in the expression levels of EPO protein in chlorella cells transformed with either of three of the transformation vectors. These data indicate that the promoters from the chlorella virus are comparable to the most common CaMV 35S promoter. Furthermore, these data suggest that other promoters from this virus can be used in future construction of chlorella transformation system for higher expression of target proteins.

• Journal of Chest Surgery
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• v.36 no.12
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• pp.921-927
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• 2003

Arterial switch operation (ASO) has been the most effective surgical option for transposition of the great arteries. But, the inappropriate dilation of the neoaortic root has been reported and its effect on neoaortic valve function and growth of aorta has not been well documented. Material and Method: Forty-eight patients who underwent cardiac catheterization during follow up after arterial switch operation were included in this study. Arterial switch operation was performed at a median age of 18 days (range 1∼211 days). Preoperative cardiac catheterization was performed in 26 patients and postoperative catheterization was performed in all patients at 15.8$\pm$9.6 months after ASO. Postoperative ratios of the diameters of neoaortic annulus, root and aortic anastomosis against the descending aorta were compared to the size of preoperative pulmonary annular, root and sinotubular junction. Preoperative and operative parameters were analyzed for the risk factors of neoaortic insufficiency. Result: There were two clinically significant neoaortic insufficiencies (grade$\geq$II/IV) during follow up, one of which required aortic valve replacement. Another patient required reoperation due to aortic stenosis on the anastomosis site. Post-operatively, neoaortic annulus/DA ratio increased from 1.33$\pm$0.28 to 1.52$\pm$.033 (p=0.01) and neoaortic root/DA ratio increased form 2.02$\pm$0.40 to 2.56$\pm$0.38 (p<0.0001). However, the aortic anastomosis/DA ratio showed no statistically significant difference (p=0.06). There was no statistically significant correlation between the occurrence of neoaortic insufficiency and neoaortic annulus/DA ratio and neoaortic root/DA ratio. Non-neonatal repair (age>30days) (p=0.02), preopeative native pulmonaic valve stenosis (p=0.01), and bisuspid pulmonic valve (p=0.03) were the risk factors for neoaortic insufficiency in univariate risk factor analysis. Conclusion: After ASO, aortic anastomosis site showed normal growth pattern proportional to the descending aorta, but neoaortic valve annulus and root were disproportionally dilated. Significant neoaortic valve insufficiency rarely developed after ASO and neoaortic annulus and root size do not correlate with the presence of postoperative neoarotic insufficiency. ASO after neonatal period, preoperative native pulmonary valve stenosis, and bicuspid native pulmonic valve are risk factors for the development of neoaortic insufficiency.

• Parasites, Hosts and Diseases
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• v.31 no.2
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• pp.149-156
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• 1993

Monoclonal antibodies (Mabs) were produced against crude scolex extract of T solium metacestodes, and applied to ELISA-inhibition test for improving the specificity of serodiagnosis of human cysticercosis. Four hybridomas secreting species-specific anti- cysticercal Mabs (Cya-1, Cya-7, Cya-28 and Cya-31) were selected. Each Mab reacted on antigenic components of 25.5 kDa (Cya-1), 28 kDa (rcya-7), 87.5 kDa (Cya-281), and 12.5 kDa (Cya-31). IFA showed that Cya-1 was located at the calcium corpuscles, and Cya-7 at the loose connective tissue of T soLium metacestode scolex. Cya-28 and Cya-31 reacted on the tegument of the scolex. By conventional ELISA, 23 out of 28 (82.1%) cysticercosis patients were found serologically positive, but 1 out of 9 (11.1%) sparganosls cases and 6 out of 31 (19.4%) paragonlmiasis cases showed false positives. By ELISA-Inhibition test using species-specific anti-cysticercal Mab Cya-7, 19 out of 28 (67.9%) cystlcercosis cases were found serologically positive, but there were no false positives In other parasitic infections.

• Parasites, Hosts and Diseases
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• v.31 no.2
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• pp.141-148
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• 1993

ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting un-n westemanl specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchlasls cases (12.5%),3 of 26 cystlcercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-Inhibition test using a R westermcni specific Mab, 100% of patagonimlasls cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-Inhlbltlon test using a Mab showed higher specificity in comparison with macro-ELISA for serodlgnosis of human paragonimlasis.

• Journal of Applied Biological Chemistry
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• v.61 no.4
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• pp.405-410
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• 2018

Pseudomonas tolaasii strain No. 6264 has been isolated from mushroom tissue and identified as one of the major pathogen causing brown blotch disease. It secretes peptide toxins, known as tolaasin and its analogue peptides. P. tolaasii 6264 has been used as a typical pathogenic strain to study the brown blotch disease for last 20 years after confirming its blotch-forming ability, hemolytic activity, and white line formation. In this study, the characteristics of P. tolaasii 6264 strain were analyzed and compared according to storage period. Strains of P. tolaasii 6264 stored annually since 2012 were cultured and their pathogenic characters were analyzed. When the 16S rRNA sequences were compared, all strains were divided into two groups. Pathogenic characters including hemolytic activity, blotch-forming ability, and white line test were also investigated. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, had all three activities; however, the rest of stored strains showed only blotch-forming ability losing other pathogenic characters. Tolaasin peptides were purified from the bacterial cultures and analyzed by mass spectrometry. The strains, P. tolaasii 6264-15-2 and P. tolaasii 6264-17, secreted Tol I (1987 Da), Tol II (1943 Da), and its analogues (1973 Da, 2005 Da) while some of these peptides were not found in the media cultured other strains. These results indicate that the pathogenicity of P. tolaasii could be varied during the storage period.