• YAKHAK HOEJI
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• v.45 no.3
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• pp.245-250
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• 2001

Aminoacyl tRNA synthetases of bacteria are known as potential targets for new anti-microbial agents. To isolate new inhibitors of bacterial methionyl-tRNA synthetases from natural sources, a new target-oriented screening system using whole cells which are over-expressing a target enzyme was developed. Approximately 8,000 culture broths of microorganisms from soils were tested by this screening system. Among them, ten culture broths was found to contain inhibitory activity against methionyl -tRNA synthetases of Escherichia coli. For the validation of the screening system, this new method was compared with in vitro enzymatic method. Seven out of 10 culture broths showed inhibitory activity against methionyl-tRNA synthetases of E. coli. This result showed that the new screening system was comparable to the enzyme assay. Thus we believe that our screening system as a new method can be applied for the screening of new antibiotics inhibiting bacterial methionyl-tRNA synthetases from natural products.

• YAKHAK HOEJI
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• v.46 no.1
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• pp.24-29
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• 2002

Enoyl-Acyl Carrier Protein Reductase (Fabl) of bacteria is hem as an important target for new antibacterial drugs and plays a determinant role in completing cycles of elongation in type-H fatty acid synthase system. In this study, a fabI gene from Staphylococcus aureus 6538p cloned in pET-l4b vector and FabI protein was over-produced in Escherichaia coli BL2l (DE3). $NH_2$-terminal His-tagged FabI protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metalaffinity chromatography Purified 6xHis-tagged FabI showed a catalytic activity on tram - 2 - octenoyl - N -acethlcysteamine by utilizing NADPH as a cofactor. For the discovery of new FabI inhibitors from chemical libraries, a target-oriented screening system using a 96-well plate was developed. About 10,000 chemical libraries from Korea Chemical Bank wore tested in this screening system, and 26 chemicals (0.25%) among them showed an inhibitory activity against FabI enzyme. This result showed that a new screening system can be used for the discovery of new FabI inhibitors.

• YAKHAK HOEJI
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• v.47 no.3
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• pp.184-189
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• 2003

Peptide deformylase (PDF) is essential and unique to bacteria, thus making it an attractive target for the discovery of novel antibacterial drugs. PDF deformylates the N-formylmethionine of newly synthesized polypeptides in prokaryotes. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and PDF protein was over-produced in Escherichia coli BL21 (DE3). NH$_2$-terminal His-tagged PDF protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. Enzymatic activity of purified 6xHis-tagged PDF was tested on the substrate (formyl-Methionine-Alanine-Serine) by formate dehydrogenase-coupled spectrometric assay of peptide deformylase. For the discovery of new PDF inhibitors from chemical libraries and culture broths of soil bacteria, a target-oriented screening system using a 96-well plate was developed. About 3,000 commercial chemical libraries were tested in this screening system, and 2 chemicals (0.07％) among them showed an inhibitory activity against PDF enzyme. This result showed that a new screening system can be used for the discovery of new PDF inhibitors.

• The Plant Pathology Journal
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• v.24 no.1
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• pp.24-30
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• 2008

Colletotrichum gloeosporioides forms an appressorium, a specialized infection structure, to infect its hosts. Among 400 and 600 culture filtrates from fungi and class Actinomycetes, six methanol extracts (A5005, A5314, A5387, A5560, A5597, and A5598) from the class Actinomycetes significantly inhibited appressorium formation in C. gloeosporioides infecting pepper fruits in a dose-dependent manner, while conidial germination was slightly enhanced. Two (A5005 and A5560) of them also exhibited distinctive inhibitory effect on the disease progress of pepper anthracnose. Water fractions of both culture filtrates also specifically inhibited appressorium formation in C. gloeosporioides and pepper anthracnose disease. Inhibition of appressorium formation by culture filtrate of A5005 was partially restored by the exogenous calcium. This results suggests that chemicals within A5005 extents its biological activity through disturbance of intracellular $Ca^{2+}$ regulation during prepenetration morphogenesis by C. gloeosporioides. Together, cell-based and target-oriented screening system used in this study should be applicable for other plant pathogenic fungi prerequisite appressorium formation to infect their hosts.

• Korean Journal of Computational Design and Engineering
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• v.17 no.1
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• pp.26-34
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• 2012

Assembly scheduling in shipbuilding is responsible for determine assembly process orders and departmental production schedule for the block, the basic composite unit of ships. It is necessary much more information to decide production scheduling as the characteristic of shipbuilding which has been more complex and more various and also, a lot of waste of time and of human power is generated in the course of data processing. The target shipbuilding manufacturer of this study use empirical techniques, based on the user's discretion, to compile and to apply data which are scattered in DB storages separately. Because of that reason, the user should not only be performed identification and screening operations but also modification and verification for vast amounts of data, so it is hard to keep the consistency of the data and also the operation time is not constant. Accordingly, the object in this study is by presenting an efficient DB framework to reduce wasting time and man-hour at experienced-oriented process, abate user's manual operations and support an efficient scheduling in assembly processes.