• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.445-449
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• 2003

As for the purpose, we first introduce an random mutation into wild-type gene to expand a mutation space, and then further recombine the mutant genes by staggered extension process PCR. As a result, we obtained the best clones 6-52 that showed a high activity and stability, from a round of error prone and staggered extension process PCR. The purified enzyme showed a similar pH stability to the wild-type enzyme and reveal a slightly high optimum pH at 12. In the optimum temperature, an identical dependency was also showed and a quite high stability in the thermal stability was obtained. Along with this, the enzyme was also stable at a reaction that supplement with a 15 % of ethanol as an additive. The addition of other solvents and surfactants did not improve the reaction and thus resulted in a similar profile to those of wild-type enzyme. The specific activity on the target compound rac-ketoprofen ethyl ester was calculated to be about 85, 000 unit, and the kinetic constants Km and Vmax were determined to be 0.2 mM and 90 mM/mg-protein/min respectively. The deduced amino acid alignment with the wild type enzyme revealed five mutations at L120P, I208V, T249A, D287H and T357A. Based on these observations, the site directed mutagenesis to delineate the mutagenic effect is under progress.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.57-63
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• 2003

Using fungal (Fusarium solani f. pisi) and bacterial (Pseudomonas mendocina) cutinases, the initial hydrolysis rate of p-nitrophenyl esters was systematically estimated for a wide range of enzyme and substrate concentrations using a 96-well microplate reader. Both cutinases exhibited a high substrate specificity; i.e. a high hydrolytic activity on p-nitrophenyl butyrate (PNB), yet extremely low activity on p-nitrophenyl palmitate (PNP). When compared to the hydrolysis of PNB and PNP by other hydrolases [lipases and esterases derived from different microbial sources, such as bacteria (Pseudomonas cepacia, Psedomonas furescens, Baciilus stearothermophilus), molds (Aspeillus niger, mucor miehei), and yeasts (Candida rugosa, Candida cylindracea)], the above substrate specificity would seem to be a unique characteristic of cutinases. Secondly, the hydrolytic activity of the cutinases on PNB appeared much faster than that of the other hydrolytic enzymes mentioned above. Furthermore, the current study proved that even when the cutinases were mixed with large amounts of other hydrolases (lipases or esterases), the Initial hydrolysis rate of PNB was determined only by the cutinase concentration for each PNB concentration. This property of cutinase activity would seem to result from a higher accessibility to the substrate PNB, compared with the other hydrolytic enzymes. Accordingly, these distinct properties of cutinases may be very useful in the rapid and easy isolation of various natural cutinases with different microbial sources, each of which may provide a novel industrial application with a specific enzymatic function.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.38-45
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• 2013

In this study, euryhaline marine microorganism, Bacillus sp. strain EBW4 isolated from polychaete (Perinereis aibuhitensis) of Suncheon Bay was physiologically, biochemically and genetically characterized. Based on 16S rRNA sequence, EBW14 was found to share 98.25% similarity with Bacillus hemicentroti $JSM076093^T$, 97.96% similarity with Bacillus hwajinponensis SW-$72^T$ and 96.28% similarity with B. algicoa $KMM3737^T$, respectively. The temperature range for the growth of strain EBW4 was $4-40^{\circ}C$, NaCl concentration range 0-17% and pH range pH 5-9, revealing that EBW4 was euryhaline bacterium. Major fatty acids in strain EBW4 were composed of anteiso $C_{15:0}$ (48.2%), iso $C_{16:0}$ (12.1%), anteiso $C_{17:0}$ (11.6%) and iso $C_{14:0}$ (9.4%). EBW4 was found to have DNase, amylase, protease and lipase for the degradation of macromolecules such as DNA, carbohydrates, proteins, lipids, etc. The enzyme activities of alkaline phosphatase, esterase (C4), leucine arylamidase and ${\alpha}$-chymotrypsin were also found in strain EBW4. Analysis of the biodegradation ability of EBW4 for organic hydrocarbons under different salinity conditions using synthetic water waste revealed that EBW4 exhibited the ability to degrade organic hydrocarbons very quickly, suggesting strain EBW4 may be a good candidate for the application to various industries.

• Korean Journal of Environmental Agriculture
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• v.14 no.3
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• pp.319-328
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• 1995

This study was carried out to investigate the resistance mechanism by three different kinds of procymidone-resistant and susceptible isolates of Botrytis cinerea which had been collected from green houses. The average resistance level of the resistant strains was 1,000 times higher than that of susceptible ones. Also, it was revealed that the resistance was not originated from components excreted by Botrytis cinerea, based on the result obtained from the treatment with piperonyl butoxide and triphenyl phosphate as an inhibitor of monooxygenase and esterase, respectively. The total lipod content of resistant strains was 1.3 times higher than that of susceptible ones, among fatty acids, palmitic acid, stearic acid, and linoleic and being 3.0, 2.5, and 2.0 times higher, respectinely. Also slight differences in sterol contents and components were observed. The crude chitin content was slightly higher in susceptible strains but contents of N-acetyl glucosamine, a hydrolysate of chitin, were about 2 times higher in resistant ones.

• Korean Journal of Environmental Agriculture
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• v.11 no.1
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• pp.1-8
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• 1992

New formulation of diazinon as granule preparing with piperonyl butoxide(PBO) and triphenyl phosphate(TPP) known as the inhibitor of monooxygenase(mo) and esterase was tested with the mortality of rice leaf hopper. Also, the mixed formulation of diazinon with tricyclazole, carbofuran and EPN were investigated about the residual pattern of diazinon in submerged soil condition at $30{\pm}2^{\circ}C$. Although the mortality about rice leaf hopper was higher only 10% on new formulation of diazinon than commercial at recommended dose but was 30-60% at half dose. Half life of diazinon in the formulation with 0.1% of PBO+TPP was 4.53 days in sterilized soil and 2.33 days in non-sterilized soil, and it was delayed about 0.74 and 0.45 respectively in contrast with the commercial granule. Also, half life of the new formulation with 1% PBO and PBO+TPP was delayed 0.44(PBO) and 0.65day(PBO+TPP) in non-sterilized soil condition, respectively. Half life of mixed formulation with tricyclazole, carbofuran, EPN or EPN+Carbofuran was delayed 2.61, 1.04, 0.43 or 2.7 days, respectively. As a result, the persistence rate of new formulation and mixed formulation was increased by inhibition of two enzymes affecting to the dgradation of diazinon in submerged soil.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.10
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• pp.1363-1368
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• 2008

The extraction and quantitative analysis conditions for astaxanthin from Haematococcus pluvialis, and the structural characteristics of H. pluvialis extract, H. pluvialis hydrolysate and synthetic astaxanthin were investigated using UV/visible and FT-IR spectrometers. Astaxanthin was dissolved in methanol, and then treated to enhance the solubility by sonication for 45 min. With sonication pretreatment, the solubility of astaxanthin increased up to 1.5 times compared to that without sonication. The extracts were hydrolyzed by cholesterol esterase for the analysis of H. pluvialis extract containing astaxanthin ester. A HPLC method using reverse phase C18 column with methanol-water (95:5, v/v) as mobile phase was developed to analyze astaxanthin. After hydrolysis, the absorption spectrum of H. pluvialis hydrolysat was changed to similar pattern to synthetic astaxanthin, confirming the extraction and analysis condition of astaxanthin from H. pluvialis.

• Korean journal of applied entomology
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• v.39 no.2
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• pp.123-129
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• 2000

This study was conducted to analyse the distribution of root-knot nematodes (Meloidogyne spp.) in fruit vegetable production areas in Korea. Soil samples were collected from greenhouses in Sungju (Kyungpook), Yeoju (Kyungki), Haman (Kyungnam), and Chungwon (Choongpook) provinces in 1997-1999. Plant parasitic nematodes were separated for density counting and some of the root-knot nematodes were identified using enzyme phenotypes of malate dehydrogenase (MDH) and esterase (EST). Among the 185 farms in Sungju province, Meloiciogyne spp. were detected from 99 farms (53.5%). Other plant parasitic nematodes detected were; Helic~otylmchuss pp. from 7 farms, Aphelenchus spp. from 43 farms, and Criconematids from 26 farms. Using the female enzyme phenotypes of MDH and EST. the four major root-knot nematodes in Korea, M. incognitc~(M I), M. uretznri~(M~ A), M. huplu (MH), and M. juvunica, could be identified. In the enzyme phenotype identification of 13 populations collected from Sunnam in Sungju province, 6 populations were identified as MA, 5 populations were identified as MI, and 2 populations were mixed with MI and MA. Among the 6 populations from Chojun in Sungju province. 4 populations were MA, one population was MI, and one population showed enzyme phenotypes of unknown species. Among the 14 populations of Yeoju province, l I populations were MH and 3 populations were MA.

• The Korean Journal of Mycology
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• v.22 no.4
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• pp.386-393
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• 1994

To investigate taxonomical relationships of Fusarium species, $esterase-{\alpha},\;-{\beta}$, acid phosphatase, malate dehydrogenase, peroxidase and polygalacturonase were extracted and the isozyme patterns were compared by using polyacrylamide gel electrophoretic analysis. Only polygalacturonase was monozyme and the other enzymes showed little differences in banding patterns. Genetic similarities based on the isozyme banding patterns were as follows: the interspecific similarity between F. subglutinans and F. moniliforme in Liseola showed the closest relationship of 74.3% of all species studies. And the similarity between section Elegans and section Liseola was 45.4%. F. napiforme and F. nygamai showed the similarity of 64.7%, similar to the correlation between species in the same section. The similarity of these two species to Liseola and Elegans showed 55.2% and 45.4%, being revealed that they would be closer to Liseola than Elegans. However, these results were similar to those of any other sections. Therefore it suggested that these 2 species should be in a different section from any other sections. And F. graminearum showed the similarity of 28.2% to the other 6 species.

• The Korean Journal of Mycology
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• v.25 no.3 s.82
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• pp.176-190
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• 1997

Sixty-three isolates of Lentinus edodes obtained from Korea were used to assess the genetic similarity by isozyme polymorphisms and random amplified polymorphic DNA patterns. The activities of esterase, peroxidase and acid phosphatase displayed 10, 7 and 3 distinct isozyme patterns, respectively. By combining the isozyme patterns obtained with the 3 enzymes, every isolate showed its own distinct electrophoretic phenotypes. A distance matrix calculated between all pairs of 63 electrophoretic phenotypes based on the presence or abscence of isozyme bands were analyzed by the group-average method. Results of the cluster analysis assinged the 63 phenotypes into six major groups. In the analysis of random amplified polymorphic DNA patterns, all isolates of Lentinus edodes were devided into five RAPD groups.

• Korean Journal of Environmental Agriculture
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• v.11 no.2
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• pp.109-115
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• 1992

In order to investigate waether soil microorganism prefer diazinon to humic substances as their substate, the growth of soil bacteria and the activities of monooxygenase and ${\alpha}-,{\beta}-esterase$ were measured after treatment of diazinon and humic substances in defined medium at $30{\pm}1^{\circ}C$. Also, the degradation rate of diazinon was determined by addition of humic substances to the medium. The number of soil bacteria was increased from 1 day and 3 days after the treatment with humic substances and diazinon, respectively. And it showed about 1.5 times more with humic substances than diazinon at 10 days. Monooxygenase(MO) activity with the treatment of humic substances was higher than diazinon until 3 days after treatment in the order of HA > FA > humin. Esterase(ES) activity with the treament of humin and HA was higher than dizainon from 5 dyas, but FA was much similar to diazinon. The degradation rate of diazinon showed more persistancy by addition of humic substances ; 51.4% with humin treatment, 58.9% with HA, 62.4% with FA and 71.9% in control at 10 days after treatment. Therefore, as soil microorganisms perfer humic substances to diazinon, the degreadation rate of diazinon might be delayed by addition of humic stbstances in submerged soil.