• Title/Summary/Keyword: enzyme specific activity

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Isolation and Mycelial Cultivation Submerged of Phellinus sp. (Phellinus sp.의 분리 및 균사체의 액체배양)

  • Kang, Tae-Su;Lee, Dong-Gi;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.257-267
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    • 1997
  • Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

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Crop Injury (Growth Inhibition) Induced by Herbicides and Remedy to Reduce It (제초제(除草劑) 약해발생(藥害發生) 양상(樣相)과 경감대책(輕減對策))

  • Kim, K.U.
    • Korean Journal of Weed Science
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    • v.12 no.3
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    • pp.261-270
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    • 1992
  • Many herbicides that are applied at the soil before weed emergence inhibit plant growth soon after weed germination occurs. Plant growth has been known as an irreversible increase in size as a result of the processes of cell divison and cell enlargement. Herbicides can influence primary growth in which most new plant tissues emerges from meristmatic region by affecting either or both of these processes. Herbicides which have sites of action during interphase($G_1$, S, $G_2$) of cell cycle and cause a subsequent reduction in the observed frequency of mitotic figures can be classified as an inhibitor of mitotic entry. Those herbicides that affect the mitotic sequence(mitosis) by influencing the development of the spindle apparatus or by influencing new cell plate formation should be classified as causing disruption of the mitotic sequence. Sulfonylureas, imidazolinones, chloroacetamides and some others inhibit plant growth by inhibiting the entry of cell into mitosis. The carbamate herbicides asulam, carbetamide, chlorpropham and propham etc. reported to disrupt the mitotic sequence, especially affecting on spindle function, and the dinitroaniline herbicides trifluralin, nitralin, pendimethalin, dinitramine and oryzalin etc. reported to disrupt the mitotic sequence, particularly causing disappearence of microtubles from treated cells due to inhibition of polymerization process. An inhibition of cell enlargement can be made by membrane demage, metabolic changes within cells, or changes in processes necessary for cell yielding. Several herbicides such as diallate, triallate, alachlor, metolachlor and EPTC etc. reported to inhibit cell enlargement, while 2, 4-D has been known to disrupt cell enlargement. One potential danger inherent in the use of soil acting herbicides is that build-up of residues could occur from year to year. In practice, the sort of build-up that would be disastrous is unikely to occur for substances applied at the correct soil concentration. Crop injury caused by soil applied herbicides can be minimized by (1) following the guidance of safe use of herbicides, particularly correct dose at correct time in right crop, (2) by use of safeners which protect crops against injury without protecting any weed ; interactions between herbicides and safeners(antagonists) at target sites do occur probably from the following mechanisms (1) competition for binding site, (2) circumvention of the target site, and (3) compensation of target site, and another mechanism of safener action can be explained by enhancement of glutathione and glutathione related enzyme activity as shown in the protection of rice from pretilachlor injury by safener fenclorim, (3) development of herbicide resistant crops ; development of herbicide-resistant weed biotypes can be explained by either gene pool theory or selection theory which are two most accepted explanations, and on this basis it is likely to develop herbicide-resistant crops of commercial use. Carry-over problems do occur following repeated use of the same herbicide in an extended period of monocropping, and by errors in initial application which lead to accidental and irregular overdosing, and by climatic influence on rates of loss. These problems are usually related to the marked sensitivity of the particular crops to the specific herbicide residues, e.g. wheat/pronamide, barley/napropamid, sugarbeet/ chlorsulfuron, quinclorac/tomato. Relatively-short-residual product, succeeding culture of insensitive crop to specific herbicide, and greater reliance on postemergence herbicide treatments should be alternatives for farmer practices to prevent these problems.

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Distinct Spatio-temporal Expression Patterns of Patatin Promoter-GUS Gene Fusion in Transgenic Potato Microtubers (형질전환 감자 소괴경의 발달단계에 따른 Patatin Promoter-GUS 유전자의 발현 분석)

  • Youm, Jung-Won;Kim, Mi-Sun;Lee, Byoung-Chan;Kang, Won-Jin;Jeon, Jae-Heung;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.13-18
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    • 2003
  • This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

The Relationship between the Salivary IgA against AgI/II of S. mutans and Dental Caries Experience among Children and Adults (소아와 성인의 타액 내 AgI/II 특이 IgA 와 우식경험도의 관계)

  • Lim, Su-Min;Kim, Jae-Gon;Baik, Byeong-Ju;Yang, Yeon-Mi;Kim, Su-Kyung;Lee, Kyung-Yol
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.4
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    • pp.671-676
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    • 2008
  • AgI/II of Streptococcus mutans(S. mutans) is an important virulence factor that contributes to the pathogenesis of S. mutans-induced dental caries. In oral cavity, salivary IgA antibodies act as safeguards against enormous challenges from oral bacteria. IgA antibodies inhibit adherence of cariogenic microorganisms to hard surfaces. Analysis of salivary IgA against AgI/II can be very useful diagnostic and powerful communication tools to the dental caries The purpose of this study was to investigate correlation between salivary AgI/II specific IgA and incidence of dental caries among children and young adults. Subjects consisted of 28 children and 18 adults. They were assigned to four groups : Group I deft index $\leq$3), Group II(deft index $\geq$4), Group III(DMFT index $\leq$3), Group IV(DMFT index $\geq$4) and they was divided two groups into caries resistant group and caries susceptible group. The study group were examined caries activity and their salivary IgA was evaluated by enzyme-linked immunosorbent assay. The results are as follows : 1. There was a positive correlation between the number of S. mutans and caries activity. 2. The titer of salivary IgA against the AgI/II was significantly higher in caries resistant group than caries susceptible group(p<0.01). 3. The titer of salivary IgA against the AgI/II in Group III was significantly higher than Group II(p<0.05).

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The Role of Heme Oxygenase-1 in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 역할)

  • Jung, Jong-Hoon;Kim, Hak-Ryul;Kim, Eun-Jung;Hwang, Ki-Eun;Kim, So-Young;Park, Jung-Hyun;Kim, Hwi-Jung;Yang, Sei-Hoon;Jeong, Eun-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.304-313
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    • 2006
  • Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.

Autoradiographic Studies on the Inhibitory Effect of Dibutyryl Cyclic AMP on Mouse Oocyte Maturation in Vitro (Dibutyryl Cyclic AMP가 생쥐여포난자의 성숙에 미치는 억제효과에 관한 자기방사법적 연구)

  • Choi, Choon-Keun
    • Applied Microscopy
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    • v.7 no.1
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    • pp.21-43
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    • 1977
  • This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

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Enzymatic Characteristics of Water-Insoluble ${\alpha}-Amylase$ Immobilized on Dithiocarbamate Wool (Dithiocarbamate Wool에 고정(固定)된 불용성(不溶性) ${\alpha}-Amylase$의 특성(特性)에 대하여)

  • Lee, Kyung-Hee;Kim, Jong-Deog;Lee, Kang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.164-170
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    • 1985
  • Yellowish modified wool, dithiocarbamate(DTC) wool, was synthesized by partial hydrolysis in 0.2 N-NaOH reacting with carbon disulfide to use as ${\alpha}-amylase$ immobilization matrix. ${\alpha}-amylase$ was immobilized reacting with sulfide group of DTC-wool by covalent binding within 1 hour. 0.5 gram of this preparation, $DTC-wool-{\alpha}-amylase$, contained 150 ug of enzyme protein and its specific activity was about 90% of the native one. General properties of $DTC-wool-{\alpha}-amylase$ were a little different from optimum temperature, optimum pH, heat stability, kinetic constants and activation energy. An apparent Michaelis constant and maximum velocity of $DTC-wool-{\alpha}-amylase$ were 5.56 mg/ml and 0.37 mg/ml. $min^{-1}$ respectively, while activation energy was 16.6 kcal/mole.

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The Heterotrimeric Kinesin-2 Family Member KIF3A Directly Binds to Creatine Kinase B (Heterotrimeric kinesin-2의 KIF3A와 creatine kinase B의 결합)

  • Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang-Jin;Lee, Won Hee;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.31 no.3
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    • pp.257-265
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    • 2021
  • Heterotrimeric kinesin-2 is a molecular motor protein of the kinesin superfamily (KIF) that moves along a microtubule plus-end directed motor protein. It consists of three different motor subunits (KIF3A, KIF3B, and KIF3C) and a kinesin-associated protein 3 (KAP3) that form a heterotrimeric complex. Heterotrimeric kinesin-2 interacts with many different binding proteins through the cargo-binding domain of the KIF3s. The activity of heterotrimeric kinesin-2 is regulated to ensure that the cargo is directed to the right place at the right time. How this regulation occurs, however, remains in question. To identify the regulatory proteins for heterotrimeric kinesin-2, we performed yeast two-hybrid screening and found a specific interaction with creatine kinase B (CKB), which is the brain isoform of cytosolic creatine kinase enzyme. CKB bound to the cargo-binding domain of KIF3A but did not interact with the KIF3B, KIF5B, or KAP3 in the yeast two-hybrid assay. The carboxyl (C)-terminal region of CKB is essential for the interaction with KIF3A. Another protein kinase, CaMKIIa, interacted with KIF3A, but GSK3a did not interact with KIF3A in the yeast two-hybrid assay. KIF3A interacted with GST-CKB-C but not with GSK-CKB-N or GST alone. When co-expressed in HEK-293T cells, CKB co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. These results suggest that the CKB-KIF3A interaction may regulate the cargo transport of heterotrimeric kinesin-2 under energy-compromised conditions in cells.

The Evaluation of IL-8 in the Serum of Pneumoconiotic patients (진폐증 환자에서의 혈청내 IL-8 농도)

  • Ahn, Hyeong Sook;Kim, Ji Hong;Chang, Hwang Sin;Kim, Kyung Ah;Lim, Young
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.945-953
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    • 1996
  • Background : Many acute and chronic lung diseases including pneumoconiosis are characterized by the presence of increased numbers of activated macrophages. These macrophages generate several inflammatory cell chemoattractants, by which neutrophil migrate from vascular compartment to the alveolar space. Recruited neutrophils secrete toxic oxygen radicals or proteolytic enzymes and induce inflammatory response. Continuing inflammatory response results in alteration of the pulmonary structure and irreversible fibrosis. Recently, a polypeptide with specific neutrophil chemotactic activity, interleukin-8(IL-8), has been cloned and isolated from a number of cells including : monocytes, macrophages and fibroblasts. IL-1 and/or TNF-${\alpha}$ preceded for the synthesis of IL-8, and we already observed high level of IL-1 and TNF-${\alpha}$ in the pneumoconioses. So we hypothesized that IL-8 may be a central role in the pathogenesis of pneumoconiosis. In order to evaluate the clinical utility of IL-8 as a biomarker in the early diagnosis of pneumoconiosis, we investigated the increase of IL-8 in the pneumoconiotic patient and the correlation between IL-8 level and progression of pneumoconiosis. Method : We measured IL-8 in the serum of 48 patients with pneumoconiosis and 16 persons without dust exposure history as a control group. Pneumoconiotic cases were divided into 3 groups according to ILO Classification : suspicious group(n=16), small opacity group(n=16) and large opacity group(n=16). IL-8 was measured by a sandwich enzytne immunoassay technique. All data were expressed as the $mean{\pm}standard$ deviation. Results: 1) The mean value of age was higher in the small opacity and large opacity group than comparison group, but smoking history was even. Duration of dust exposure was not different among 3 pneumoconiosis groups. 2) IL-8 level was $70.50{\pm}53.63pg/m{\ell}$ in the suspicious group, $107.50{\pm}45.88pg/m{\ell}$ in the small opacity group, $132.50{\pm}73.47pg/m{\ell}$ in the large opacity group and $17.85{\pm}33.85pg/m{\ell}$ in the comparison group. IL-8 concentration in all pneumoconiosis group was significant higher than that in the comparison group(p<0.001). 3) IL-8 level tended to increase with the progression of pneumoconiosis. Multiple comparison test using Anova/Scheffe analysis showed a significant difference between suspicious group and large opacity group(p<0.05). 4) The level of IL-8 was correlated with the progression of pneumoconiosis(r=0.4199, p<0.05). Conclusion : IL-8 is thought to be a good biomarker for the early diagnosis of pneumoconiosis.

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