• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1655-1660
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• 2007

A metagenome is a unique resource to search for novel microbial enzymes from the unculturable microorganisms in soil. A forest soil metagenomic library using a fosmid and soil microbial DNA from Gwangneung forest, Korea, was constructed in Escherichia coli and screened to select lipolytic genes. A total of seven unique lipolytic clones were selected by screening of the 31,000-member forest soil metagenome library based on tributyrin hydrolysis. The ORFs for lipolytic activity were subcloned in a high copy number plasmid by screening the secondary shortgun libraries from the seven clones. Since the lipolytic enzymes were well secreted in E. coli into the culture broth, the lipolytic activity of the subclones was confirmed by the hydrolysis of p-nitrophenyl butyrate using culture supernatant. Deduced amino acid sequence analysis of the identified ORFs for lipolytic activity revealed that 4 genes encode hormone-sensitive lipase (HSL) in lipase family IV. Phylogenetic analysis indicated that 4 proteins were clustered with HSL in the database and other metagenomic HSLs. The other 2 genes and 1 gene encode non-heme peroxidase-like enzymes of lipase family V and a GDSL family esterase/lipase in family II, respectively. The gene for the GDSL enzyme is the first description of the enzyme from metagenomic screening.

• Journal of Plant Biotechnology
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• v.30 no.2
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• pp.207-214
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• 2003

This study was carried out to establish a novel bioassay system for screening of compounds affecting aromatic amino acid or anthocyanin biosynthesis through investigating a degree of sucrose-induced anthocyanin formation such as size of plant material, buffer conditions, light intensity and irradiated duration, incubation temp., etc were determined and standard procedure (suitable experimental condition) was set up as follows. The second leaf blade of white corn seeding induced by fluridone treatment were segmented into a size of 5${\times}$5 min. The segments were floated on the solution of 1% sucrose in 1.0mM MES buffer (pH6.0∼6.5) and incubated at 26$^{\circ}C$ for 2days under the continuous light condition(70∼100$\mu$mol m$^{-2}$ s$^{-1}$ ). Anthocyanin in the purpled tissues was extracted with methanol containing 1% HCl and the optical density of the clear supematants was determined at 528mm. Influences of some chemicals were tested using this system. Glyphosate, 5-enolpyruvylshikimate 3-phosphate synthase inhibitor, showed most sensitive response with I$_{50}$ value at 3.3$\mu$M. Dicyclohexylcarbodiimide(DCCD) and parachloromercuribenzenesulfonic acid(PCMBS) had a relatively strong ingibition with I50 value at 7.1$\mu$M and 10.2$\mu$M, respectively. These results show that sucrose-induced anthocyanin formation in white com leaf segment provide a very simple and rapid system for searching new compounds affecting aromatic amino acid or anteocyanin biosynthesis by screening at less than 10$\mu$M.

• Mycobiology
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• v.48 no.4
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• pp.326-329
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• 2020

Valuable natural compounds produced by a variety of microorganisms can be used as lead molecules for development of new agrochemicals. Furthermore, high-throughput in vitro screening systems with specific modes of action can increase the probability of discovery of new fungicides. In the current study, a rapid assay tested with various microbes was developed to determine the degree of respiratory inhibition of Saccharomyces cerevisiae in two different liquid media, YG (containing a fermentable carbon source) and NFYG (containing a non-fermentable carbon source). Based on this system, we screened 100 fungal isolates that were classified into basidiomycetes, to find microbial secondary metabolites that act as respiratory inhibitors. Consequently, of the 100 fungal species tested, the culture broth of an IUM04881 isolate inhibited growth of S. cerevisiae in NFYG medium, but not in YG medium. The result is comparable to that from treatment with kresoxim-methyl used as a control, suggesting that the culture broth of IUM04881 isolate might contain active compounds showing the inhibition activity for respiratory chain. Based on the assay developed in this study and spectroscopic analysis, we isolated and identified an antifungal compound (-)-oudemansin A from culture broth of IUM04881 that is identified as Oudemansiella venosolamellata. This is the first report that (-)-oudemansin A is identified from O. venosolamellata in Korea. Taken together, the development of this assay will accelerate efforts to find and identify natural respiratory inhibitors from various microbes.

• Bulletin of the Korean Chemical Society
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• v.28 no.6
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• pp.947-952
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• 2007

Mycobacterium tuberculosis is a pathogen responsible for 2-3 million deaths every year worldwide. The emergence of drug-resistant and multidrug-resistant tuberculosis has increased the need to identify new antituberculosis targets. Acetohydroxy acid synthase, (AHAS, EC 2.2.1.6), an enzyme involved in branched-chain amino acid synthesis, has recently been identified as a potential anti-tuberculosis target. To assist in the search for new inhibitors and “receptor-based” design of effective inhibitors of tubercular AHAS (TbAHAS), we constructed four different structural models of TbAHAS and used one of the models as a target for virtual screening of potential inhibitors. The quality of each model was assessed stereochemically by PROCHECK and found to be reliable. Up to 89% of the amino acid residues in the structural models were located in the most favored regions of the Ramachandran plot, which indicates that the conformation of each residue in the models is good. In the models, residues at the herbicide-binding site were highly conserved across 39 AHAS sequences. The binding mode of TbAHAS with a sulfonylurea herbicide was characterized by 32 hydrophobic interactions, the majority of which were contributed by residue Trp516. The model based on the highest resolution X-ray structure of yeast AHAS was used as the target for virtual screening of a chemical database containing 8300 molecules with a heterocyclic ring. We developed a short list of molecules that were predicted to bind with high scores to TbAHAS in a conformation similar to that of sulfonylurea derivatives. Five sulfonylurea herbicides that were calculated to efficiently bind TbAHAS were experimentally verified and found to inhibit enzyme activity at micromolar concentrations. The data suggest that this time-saving and costeffective computational approach can be used to discover new TbAHAS inhibitors. The list of chemicals studied in this work is supplied to facilitate independent experimental verification of the computational approach.

• 한국가스학회:학술대회논문집
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• 2003.05a
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• pp.246-251
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• 2003

• Current Research on Agriculture and Life Sciences
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• v.31 no.1
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• pp.1-10
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• 2013

The exposure of food to ionizing radiation has been recognized as a safe and effective mode of food preservation in more than 55 countries. The benefits include eradication of insect pests, inactivation of food pathogens, extension of shelf-life, and improvement in food hygiene. Regulatory authorities around the world have emphasized the implementation of various national and international regulations to facilitate trade and development of consumers' confidence in purchasing irradiated foods. Therefore, the need for reliable irradiation detection methods has increased to enforce these regulations. At present, a number of promising analytical approaches have been developed and evaluated. Moreover, about 10 European Standards have been adopted as General CODEX Alimentarius methods for the detection of irradiated foodstuffs. However, most of these methods demand relatively expensive equipment and prolonged sample preparation. Therefore, simple and cost-effective approaches would be advantageous for rapid screening of foodstuffs. The suspected samples need to be analyzed further with more validated techniques to confirm the screening results. In this review, existing screening methods (i.e. physical, chemical, and biological) for the identification of irradiated foods have been outlined along with their principles, scopes and limitations.

• Journal of Environmental Health Sciences
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• v.48 no.5
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• pp.272-281
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• 2022

Background: The constant consumption of chemical products owing to expanding industrialization has led to an increase in public interest in chemical substances. As the production and disposal processes for these chemical products cause environmental problems, regional information on the hazard level of chemical substances is required considering their effects on humans and in order to ensure environmental safety. Objectives: This study aimed to identify hazard contribution and spatiotemporal characteristics by region and chemical by calculating a hazard-based result score using pollutant release and transfer register (PRTR) data. Methods: This study calculated the chemical discharge and hazard-based result score from the Risk-Screening Environmental Indicators (RSEI) model, analyzed their spatiotemporal patterns, and identified hotspot areas where chemical discharges and high hazard-based scores were concentrated. The amount of chemical discharge and hazard-based risk scores for 250 cities and counties across South Korea were calculated using PRTR data from 2011 to 2018. Results: The chemical discharge (high densities in Incheon, Daegu, and Busan) and hazard-based result scores (high densities in Incheon, Chungcheongnam-do, and some areas of Gyeongsangnam-do Province) showed varying spatial patterns. The chemical discharge (A, B) and hazard-based result score (C, D) hotspots were identified. Additionally, identification of the hazard-based result scores revealed differences in the type of chemicals contributing to the discharge. Ethylbenzene accounted for ≥80% of the discharged chemicals in the discharge hotspots, while chromium accounted for >90% of the discharged chemicals in the hazard-based result score hotspots. Conclusions: The RSEI hazard-based result score is a quantitative indicator that considers the degree of impact on human health as a toxicity-weighted value. It can be used for the management of industries discharging chemical substances as well as local environmental health management.

• Bulletin of the Korean Chemical Society
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• v.29 no.5
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• pp.921-927
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• 2008

Discovery of $\alpha$-glucosidase inhibitors has been actively pursued with the aim to develop therapeutics for the treatment of diabetes and the other carbohydrate mediated diseases. As a method for the discovery of new novel inhibitors of $\alpha$-glucosidase, we have addressed the performance of the computer-aided drug design protocol involving the homology modeling of $\alpha$-glucosidase and the structure-based virtual screening with the two docking tools: FlexX and the automated and improved AutoDock implementing the effects of ligand solvation in the scoring function. The homology modeling of $\alpha$-glucosidase from baker’s yeast provides a high-quality 3-D structure enabling the structure-based inhibitor design. Of the two docking programs under consideration, AutoDock is found to be more accurate than FlexX in terms of scoring putative ligands to the extent of 5-fold enhancement of hit rate in database screening when 1% of database coverage is used as a cutoff. A detailed binding mode analysis of the known inhibitors shows that they can be stabilized in the active site of $\alpha$- glucosidase through the simultaneous establishment of the multiple hydrogen bond and hydrophobic interactions. The present study demonstrates the usefulness of the automated AutoDock program with the improved scoring function as a docking tool for virtual screening of new $\alpha$-glucosidase inhibitors as well as for binding mode analysis to elucidate the activities of known inhibitors.

• Journal of Korean Institute of Industrial Engineers
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• v.43 no.2
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• pp.127-134
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• 2017

For product or process design and development, it is common to optimize multiple responses (characteristics) based on experimental data. To determine optimal conditions, we need to design the experiment, estimate a proper model for each response, and optimize the multiple responses simultaneously. There are several techniques and many research results on optimizing multiple responses simultaneously, when the experimental data are available. However, the experimental design issue for optimizing multiple responses has not been discussed yet. This paper proposes some idea on how to plan screening design when requirements for multiple performance characteristics are to be met in developing new products. A screening design procedure is developed for securing the requirements of multiple responses. Initial design factors are classified into three categories; specific, non-conflicting common, and conflicting common. After screening experiments, follow-up design region search method is suggested with respect to the most unsatisfied or important response, or overall desirability. A case study on a synthesis of melamine formaldehyde resin is presented to illustrate the procedure and to show the validity of the approach.

• Research in Plant Disease
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• v.18 no.2
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• pp.86-92
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• 2012

Clubroot caused by Plasmodiophora brassicae Woron. is one of the most important diseases in Brassica crops worldwide. To establish more simple and reliable screening method for resistant cabbage and broccoli to P. brassicae, the development of clubroot on the plants according to inoculum concentration and incubation period after inoculating with the pathogen was investigated using P. brassicae GN1 isolate (race 9). To facilitate and acquire precise result of resistance screening of cabbage and broccoli to clubroot, 14-day-old seedlings were inoculated by drenching roots with the spore suspension of P. brassicae to give inoculum density of $2.5{\times}10^9$ spores/pot. To develop the disease, the inoculated seedlings were incubated in a growth chamber at $20^{\circ}C$ for 3 days, and then cultivated in a greenhouse ($20{\pm}5^{\circ}C$) for five weeks. Under the optimum conditions, 16 cabbage and 17 broccoli cultivars were tested for resistance to four field isolates (GN1, GN2, GS and YC) of P. brassicae collected from four regions in Korea. Among them, some cabbage and broccoli cultivars showed different resistance response to three isolates (GN1, GN2 and GS) determined as race 9 by using the differential varieties of Williams. On the other hand, all the tested cultivars were highly susceptible to YC isolate (race 2). The results suggest that this method is efficient screening method of cabbage and broccoli for resistance to P. brassicae.