• Korean Journal of Food Science and Technology
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• v.10 no.2
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• pp.209-214
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• 1978

Naringinase produced from Aspergillus nidulans was immobilized in acrylamide gel by the entrapping method and its characteristics were studied. Optimum acrylamide concentration was 10%, but N.N'-methylene bisacrylamide concentration had no influence on the final enzyme gel activity. The suitable amount of enzyme dissolved in the polymerization reaction mixture was 126 units/ml. Optimum pH of immobilized enzyme was 5.0 which was the same as that of free enzyme. However, immobilized enzyme showed a higher optumum reaction temperature, markedly increased pH and temperature stability. In a packed-column reactor, the observed reaction rate was increased proportionally to flow rate up to 5ml/min., but independent above 6ml/min.. Activation energy of the immobilized enzyme was 13.01 Kcal/mole, and the energy required for the thermal inactivation was 39.4 Kcal/mole. The apparent Km for 100 mesh gel was $7.23{\times}10^{-3}$ mole.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.454-460
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• 2007

Enzymatic properties and substrate specificity of recombinant $\beta-glycosidases$ from a hyperthermophilic archaeon, Sulfolobus shibatae (rSSG), were analyzed. rSSG showed its optimum temperature and pH at $95^{\circ}C$ and pH 5.0, respectively. Thermal inactivation of rSSG showed that its half-life of enzymatic activity at $75^{\circ}C$ was 15 h whereas it drastically decreased to 3.9 min at $95^{\circ}C$. The addition of 10 mM of $MnCl_2$ enhanced the hydrolysis activity of rSSG up to 23% whereas most metal ions did not show any considerable effect. Dithiothreitol (DTT) and 2-mercaptoethanol exhibited significant influence on the increase of the hydrolysis activity of rSSG rSSG apparently preferred laminaribiose $(\beta1\rightarrow3Glc)$, followed by sophorose $(\beta1\rightarrow2Glc)$, gentiobiose $(\beta1\rightarrow6Glc)$, and cellobiose $(\beta1\rightarrow4Glc)$. Various. intermolecular transfer products were formed by rSSG in the lactose reaction, indicating that rSSG prefers lactose as a good acceptor as well as a donor. The strong intermolecular transglycosylation activity of rSSG can be applied in making functional oligosaccharides.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.157-167
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• 2000

A thermostable chitosanase gene from the isolated strain, Bacillus sp. CK4, was cloned, and its complete DNA sequence was determined. The thermostable chitosanase gene was composed of an 822-bp open reading frame which encodes a protein of 242 amino acids and a signal peptide corresponding to a 30 kDa enzyme in size. The deduced amino acid sequence of the chitosanase from Bacillus sp. CK4 exhibits 76.6%, 15.3%, and 14.2% similarities to those from Bacillus subtilis, Bacillus ehemensis, and Bacillus circulans, respectively. C-terminal homology analysis shows that Bacillus sp. CK4 belongs to the Cluster III group with Bacillus subtilis. The size of the gene was similar to that of a mesophile, Bacillus subtilis showing a higher preference for codons ending in G or C. The functional importance of a conserved region in a novel chitosanase from Bacillus sp. CK4 was investigated. Each of the three carboxylic amino acid residues were changed to E50D/Q, E62D/Q, and D66N/E by site-directed mutagenesis. The D66N/E mutants enzymes had remarkably decreased kinetic parameters such as $V_{max}$ and k$\sub$cat/, indicating that the Asp-66 residue was essential for catalysis. The thermostable chitosanase contains three cysteine residues at position 49, 72, and 211. Titration of the Cys residues with DTNB showed that none of them were involved in disulfide bond. The C49S and C72S mutant enzymes were as stable to thermal inactivation and denaturating agents as the wild-type enzyme. However the half-life of the C211S mutant enzyme was less than 60 min at 80$^{\circ}C$, while that of the wild type enzyme was about 90 min. Moreover, the residual activity of C211S was substantially decreased by 8 M urea, and fully lost catalytic activity by 40% ethanol. These results show that the substitution of Cys with Ser at position 211 seems to affect the conformational stability of the chitosanase.

• Korean journal of applied entomology
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• v.20 no.4 s.49
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• pp.191-195
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• 1981

Cucurbits including pumpkin (Cucurbita pepo), gourd (Lagenariaa siceraria), cucumber (Cucumis sativus), melon(Cucumis melo) and watermelon(Cucurbita anguria) were diseased with mosaic symptoms. The causal virus was identified as watermelon mosaic virus(WMV). The WMV was transmitted by Myzus persicae Sulzer, and no seed borne virus was found. The virus caused large local lesions on the inoculated leaves of the Chenopodium amaranticolor and mosaic symptom on the upper leaves of Cucumis melo, Cucumis sativus, Lagenaria siceraria, Cucurbita anguria and Cucurbita pepo. There were no symptoms on the inoculated leaves of the Nicotiana tabacum var. Bright yellow, Nicotiana glutinosa, Vigna unguiculata. Petunia hybrida and Datura stramonium. Thermal inactivation point was $55\~65^{\circ}C$, dilution end point was $10^{-4}\;10^{-5}$ and longevity in vitro of the virus was $7\~8$ days. The virus showed positive reaction against watermelon mosaic virus antiserum in microprecipitin tests. The virus particles were flexuous rods in size of 750 nm.

• Korean journal of applied entomology
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• v.19 no.2 s.43
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• pp.67-72
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• 1980

A mosaic virus disease of ginger plant was investigated to determine its virus group on the basis of host range, physical and chemical properties, serological behavior and electron-microscopic morphology. The disease gave rise to yellowsih and dark-green mosaic on the leaves in the early stage and stunted all the leaves as well as rhizomes in the late stage. In the field about 43\% of the plants were observed to be diseased The disease was able to be artificially infected to the ginger plants by the sap and transmission as well as to 23 other species of plants which were known to be the CMV susceptible plants by the sap transmission; Chenopodium amaranticolar, Nicotiana tabaccum var. Havana, cow pea, cucumber, tomato,... etc. The dilution end point of the virus ranged $10^{-4}-10^{-5}$ and the thermal inactivation point $65-70^{\circ}C$. Serological test showed a positive reaction by a CMV antiserum. An electron microscopy of the purified virus showed that the virus particles were spherical with a diameter of $28-32m\mu$. Virus particles from the infected tissue were observed to be free or aggregated in the mesophyll tissue of artificially infected tobacco plant. The mosaic disease of ginger plants were conclusively suggested to the CMV group.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.155-160
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• 2006

A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

• Korean Journal Plant Pathology
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• v.2 no.1
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• pp.48-52
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• 1986

The virus isolated from white clover, Trifolium repens showing mosaic symptom was identified as bean yellow mosaic virus (BYMV) based on the host range, physical properties, aphid transmission, serology and morphology of the virus particles. Chenopodium amaranticolor and C. quinoa produced local lesions on the inoculated leaves and chlorotic spot on the upper leaves. Broad bean and cowpea produced local lesions on the inoculated leaves and mosaic with vein necrotic symptoms on the upper leaves. French bean showed vein necrosis on the inoculated leaves, yellow mosaic on the upper leaves and bud blight. The average size of virus particles was 740nm in length. The virus was also transmitted by Myzus persicae. The thermal inactivation point of the virus isolate was $60\;to\;65^{\circ}C$, the dilution end point $10^{-3}\;-\;10^{-4}$ and the longevity in vitro was 3 days Serological tests with the virus purified from Trifolium repens were positive to BYMV antiserum.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.757-763
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• 1999

The metal specificity of D-xylose isomerase from Streptomyces rubiginosus was examined by site-directed mutagenesis. The activation constants for metal ion ($Mg^{2+},{\;}Mn^{2+},{\;}or{\;}Co^{2+}$) of wild-type and mutant enzymes were determined by titrating the metal ion-free enzyme with $Mg^{2+},{\;}Mn^{2+},{\;}and{\;}Co^{2+}$, respectively. Substitutions of amino acids either on coordinated or around the M2 site (His-22O, Asn-185, Glu-186, and Glu-221) dramatically affected the activation constants as well as activity. A decrease of metal binding affinity was most significant in the presence of $Mg^{2+}$. When compared with the wild-type enzymes, the binding affinity of H220S and Nl85K for Mg^{2+} was decreased by 10-15-fold, while the affinity for $Mn^{2+}{\;}or{\;}Co^{2+}$ only decreased by 3-5-fold. All the mutations close to the M2 site changed their metal preference from $Mg^{2+}{\;}to{\;}Mn^{2+}{\;}or{\;}Co^{2+}$. These altered metal preferences may be caused by a relatively weak binding affinity of $Mg^{2+}$ to the enzyme. Thermal inactivation studies of mutants at the M2 site also support the importance of the M2 site geometry for metal specificity as well as the thermostability of the enzyme. Mutations of other important groups hardly affected the metal preference, although pronounced effects on the kinetic parameters were sometimes observed. This study proposes that the metal specificity of D-xylose isomerase can be altered by the perturbation of the M2 site geometry, and that the different metal preference of Group I and GroupII D-xylose isomerases may be caused by nonconserved amino acid residues around the M2 site.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.542-549
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• 2010

In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

• Journal of Korean Society for Quality Management
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• v.46 no.1
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• pp.169-188
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• 2018

Purpose: The purpose of this study is to analyze low-temperature starting performance of the light attacker and to search and improve the aircraft system including battery and Battery Charge and Control Unit(BCCU). Methods: In order to improve the starting up performance of the light attacker at low-temp, various deficiency cause were derived and analyzed using Fault Tree Analysis method. As a result, it was confirmed there were drawbacks in the charging and discharging mechanism of the battery. The inactivation of the battery's electrolyte at low-temp and the premature termination of the battery charge were the main cause. After long error and trial, we improved these problems by improving performance of battery and optimizing the charging algorithm of BCCU. Results: It was confirmed that the problems of starting up failures were solved through the combined performance test of the battery and BCCU, the ground test using the aircraft system and the operation test conducted by Korea Airforce operating unit for 3 months in winter. Conclusion: This study showed that the improvement of quality reliability was achieved and thus the start-up performance issue of the light attacker has been resolved at low temperature. And it is expected that the design methodologies of temperature-affected electrical system of aircraft will contribute to the development of the aircraft industry in the future.