• 한국군사과학기술학회지
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• 제14권4호
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• pp.710-718
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• 2011

Edema factor(EF) is a portion of anthrax toxin which produces edema when combined with protective antigen. This paper describes about technique for cloning, expression, purification and activity test of EF. Using the E. coli expression system, we could make recombinant EF protein although it's origin is Bacillus anthracis. And also we could culture massively and purify highly pure protein. Finally we confirm a enzyme activity of purified EF to increase intracellular cAMP level. Through establishing this technique, it can be possible to research about EF in depth and apply to expression and purification of many other protein in biology.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1908-1914
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• 2008

An alkaliphilic bacterium, Bacillus clausii SKAL-16, was isolated from soil that had been contaminated with vegetable oil. The optimal pH and general pH range for bacterial growth was 8, and 7 to 10, respectively. The bacterium could grow on tributyrin and glycerol, but could not grow on acetate and butyrate. The SKAL-16 strain excreted butyric acid during growth on tributyrin, and selectively ingested glycerol during growth on a mixture of butyric acid and glycerol. The SKAL-16 generated intracellular lipase, but did not produce esterase and extracellular lipase. The DNA fragment amplified with the chromosomal DNA of SKAL-16 and primers designed on the basis of the esterase-coding gene of Bacillus clausii KSM-KI6 was not identical with the esterase-coding gene contained in the GenBank database. Pyruvate dehydrogenase, isocitrate dehydrogenase, and malate dehydrogenase activities were detected in the cell-free extract (crude enzyme).

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.179-182
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• 2001

Two intracellular enzymes, cellobiose phosphorylase (CBP) and cellodextrin phosphorylase (CDP) are involved in the phosphorolytic pathway in cellulose degradation. Those enzymes are considered to be useful in syntheses of oligosaccharides because the reactions are reversible. CBP from Cellvibrio gilvus and CDP from Clostridium thermocellum YM-4 were cloned and over-expressed in Escharichia coli. Both the enzyme reactions showed ordered bi bi mechanism. Acceptor specificity of CBP in the reverse reaction was determined. Several $\beta$-l,4-glucosyl disaccharides were synthesized by using the reaction. A new substrate inhibition pattern, competitive substrate inhibition, was also found in the reverse reaction of CBP Cellobiose was produced from sucrose at a high yield by a combined action of three enzymes including CBP

• 한국식품저장유통학회지
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• 제1권2호
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• pp.107-116
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• 1994

This paper was carried out to Investigate changes in the activities of cell-degrading enzymes, cell wall components and cell structure of peach during maturation and storage for valuation of quality. The firmness of peach was decreased during maturation and storage, and was remarkably decreased in Daegubo than Yumyung. Polygalacturonase and $\beta$-galactosidase activities of peach were increased during maturation and storage, and were remarably increased in soft peach and in mature and soft peach, respectively. Contents of alcohol-insoluble substance, cell wall, and total and insoluble pectin of peach were decreased during maturation and storage, but cellulose and soluble pectin were increased. Intracellular space was enlarged during maturation and middle lamella was gradually degraded during maturation.

• Archives of Pharmacal Research
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• 제27권12호
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• pp.1258-1262
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• 2004

Phosphodiesterase (PDE) 4 is an enzyme that degrades intracellular cAMP. In the present study, the effect of rolipram, a specific phosphodiesterase (PDE) 4 inhibitor, on osteoclast formation was investigated. Rolipram induced osteoclast formation in cocultures of mouse bone marrow cells and calvarial osteoblasts. This activity was not observed in the absence of calvarial osteoblasts, suggesting that calvarial osteoblasts are likely target cells of rolipram. Osteoclast formation by rolipram was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for the osteoclast differentiation factor, TNF-related activation-induced cytokine (TRANCE, identical to RANKL, ODF, and OPGL). Northern blot analysis revealed the effect of rolipram to be associated with the increased expression of TRANCE mRNA in mouse calvarial osteoblasts. Collectively, these data indicate that PDE4 inhibitor up-regulates the TRANCE mRNA expression in osteoblasts, which in turn controls osteoclast formation.

• 미생물학회지
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• 제27권1호
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• pp.27-34
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• 1989

In order to evaluate the applicability of enzyme electrophoresis for the identification of intra/interspecific hybride obtained by the protoplast fusion in Trichoderma, soluble proteins, intracellular soluble enzymes and extracellular $\beta$-glucosidase were analyzed by polyacrylamide gel electrophorsis. As the results, patterns of soluble protein, and isozyme patterns of peroxidase, malate dehydrogenase, and $\beta$-glucosidase in hydrids were defferent from those in parental and wild type strains. Therefore, it was established that the analysis of protein pattern by electrophoresis could be applied to isolate and identify the hybrids from the protoplast fusion.

• 대한화장품학회지
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• 제27권2호
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• pp.57-58
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• 2001

Biosurfactant has antibiological activities. Due to its low toxicity, biosurfactant can be applied to human health care products. Posseing the antibiological activities as well as the surfactant properties implies that biosurfactant can be widely used in the cosmetic industry. In this presentation, applications of biosurfactant have been reviewed including the antimicrobial activities of sophorolipid. Sophorolipid, a biosurfactant produced from Candida bombicola ATCC 22214, showed antimicrobial activity against Bacillus subtilis, Staphylococcus xylosus, Streptococcus mutans, and Propionibacterium acne at 4, 1, 1, 0.5 ppm as MIC (minimum inhibitory concentration). Also 100 ppm of sophorolipid inhibited 50% of cell growth of plant pathogenic fungus, B. cinera. However, sophorolipid showed no effect on the Escherichia coli., indicating its selective antimicrobial activity depending on the cell wall structure. Treatment of B. subtilis with sophorolipid increased the leakage of intracellular enzyme, malate dehydrogenase, indicating the possible interaction of sophorolipid with cellular membrane. Between lactone-type and acid-type sophorolipid, the former showed higher antimicrobial activity.

• 대한예방한의학회지
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• 제12권2호
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• pp.197-206
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• 2008

The inhibition of osteoblast by glucocorticoid is recognized as its action mechanism of decreased bone formation. In this study, the effect of JY, Jahyulyangkeuntang, on the differentiation and mineralization of osteoblastic cells was investigated in the presence of dexamethasone. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, alkaline phosphatase(ALP) activity, bone martrix production, and cell apoptosis. JY enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracellular collagen synthesis were increased when the cells were treated with JY. And JY restored calvarial cell function decreased by dexamethasone.

• 한국식품영양학회지
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• 제7권4호
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• pp.259-266
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• 1994

The hyperthermophilic archaebacterium Thermococcus profundus Isolated from a deep-sea hydrothermal vent system, produced several amylolytic enzymes such as extracellular amylase and pullulanase, intracellular a-1,4-91ucosidase in respone to the presence of complex carbohydrates In the growth medium. This strain showed high activities on 0.5% maltose than on complex carbohydrates One of the amylases was partially purified by ammonium sulfate precipitation, DEAE-Toyopearl chromatography. The amylase exhibited maximal activity at pH 5.5 and 80$^{\circ}C$, and was stable in the range of pH 5.5 to 9.5 and up to 80$^{\circ}C$ for 30 min. The enzyme activity was no dependence on Ca2+ and not inhibited by detergents. The amylase hydrolyzed soluble starch, amylose, amylopectin and glycogen to produce maltose and maltotriose with trace amounts of glucose, but not pullulan and ${\alpha}$-, ${\beta}$-, ${\gamma}$-cyclodextrin. Malto-oligosaccharides ranging from maltotetraose to maltoheptaose were hydrolyzed in an endo fashion.

• Mycobiology
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• 제47권1호
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• pp.112-119
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• 2019

Compounds from Lingzhi has been demonstrated the ability for inhibiting tyrosinase (a key enzyme in melanogenesis) activity. In this study, we investigated the anti-melanogenic activity from the submerged mycelial culture of Ganoderma weberianum and elucidated the skin lightening mechanism by B16-F10 murine melanoma cells. From the cellular context, several fractionated mycelium samples exhibited anti-melanogenic activity by reducing more than 40% extracellular melanin content of B16-F10 melanoma cells. In particular, the fractionated chloroform extract (CF-F3) inhibited both secreted and intracellular melanin with the lowest dosage (25 ppm). Further analysis demonstrated that CF-F3 inhibited cellular tyrosinase activity without altering its protein expression. Taken together, our study has demonstrated that the chemical extracts from submerged mycelial culture of G. weberianum have the potential to serve as an alternative anti-melanogenic agent.