• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.968-973
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• 2010

In order to study its biochemical properties, the integrase (IN) protein of feline foamy virus (FFV) was overexpressed in Escherichia coli, purified by two-step chromatography, (Talon column and heparin column), and characterized in biochemical aspects. For the three enzymatic reactions of the 3'-processing, strand transfer, and disintegration activities, the $Mn^{2+}$ ion was essentially required as a cofactor. Interestingly, $Co^{2+}$ and $Zn^{2+}$ ions were found to act as effective cofactors, whereas other transition elements such as $Ni^{2+}$, $Cu^{2+}$, $La^{3+}$, $Y^{3+}$, $Cd^{2+}$, $Li^{1+}$, $Ba^{2+}$, $Sr^{2+}$, and $V^{3+}$ were not. Regarding the substrate specificity, FFV IN has low substrate specificities as it cleaved in a significant level prototype foamy virus (PFV) U5 LTR substrate as well as FFV U5 LTR substrate, whereas PFV IN did not. Finally, the 3'-processing activity was observed in high concentrations of several solvents such as CHAPS, glycerol, Tween 20, and Triton X-100, which are generally used for dissolution of chemicals in inhibitor screening. Therefore, in this first report showing its biochemical properties, FFV IN is proposed to have low specificities on the use of cofactor and substrate for enzymatic reaction as compared with other retroviral INs.

• Applied Biological Chemistry
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• 제41권4호
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• pp.235-240
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• 1998

The myristoyl-acyl carrier protein (ACP) specific thioesterase from Iris tectorum was purified to a considerable homogeneity and characterized. The enzyme was eluted with a considerable stability by double-gradients using Triton X-100 and low ionic KCl or Na-phosphate through DEAE-52, Octyl-Sepharose, Q-Sepharose, and hydroxyapatite chromatoraphy. SDS-PAGE analysis showed a single band of 39 kDa. The native molecular weight was estimated to be 82 kDa by Sephacryl S-200 chromatography, indicating that the enzyme was a dimer. The thioesterase showed a chain-length specificity to myristoyl-ACP in preference to other-ACPs. The enzyme activity decreased by 1.0 mM myristate to about 27% of the original activity, whereas the remaining activity with decanoate was about 90%. The purified thioesterase was inhibited by myristoyl-CoA more than by myristate, suggesting that the myristoyl-AGP thiolesterase might be controlled by myristic acid and/or a subsequent product myristoyl-CoA. In addition, some biochemical characteristics of the enzyme were described.

• Journal of Plant Biology
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• 제32권4호
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• pp.265-274
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• 1989

The effect of Ca2+ on auxin-induced ehtylene production in etiolated mungbean (Vigna radiata W.) hypocotyls was studied. Auxin-induced ethylene production by mungbean seedlings which had been germinated in the presence of 5-10mM Ca2+ (High Ca2+ ; HC) is greater than that by seedlings which had been germinated in distilled water (Low Ca2+ ; LC). The effect of Ca2+ on auxin-induced ethylene production was greatly increased after 12hr of incubation period. The stimulation of auxin-induced ethylene production by Ca2+ was specific, since divalent cations, such as Mg2+ and Mn2+ did not enhance auxin-induced ethylene production. Calcium also promoted ethylene evoluation induced by methionine and 1-Aminocyclopropane-1-carboxylic acid(ACC). The effect of Ca2+ on auxin-induced ethylene production was not caused by increase in free IAA or ACC contents of hypocotyl tissue. Dimethyl sulfoxide and Triton X-100, that disrupts the emembranes, inhibited ethylene production to a greater extent in LC segments than in HC segments. Addition of Ca2+ to the incubation medium for LC segments resulted in enchancement of ethylene production probalby because the membrane integrity is supported under these conditions. Comparison of activity of Ethylene Forming Enzyme(EFE) in LC and HC hypocotyl segments indicated that the enzyme activity of HC was about 2 times higher than that of L.C. It is suggested that Ca2+ increases the activity of plasma membrane-bound EFE through its stabilizing effect onn the membrane, which in turn brings about promotion of ethylene production.

• 한국환경과학회지
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• 제18권1호
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• pp.15-19
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• 2009

Early studies claimed that heterotrophic dinoflagellates Pfiesteria piscicida and related genera may produce a putative water-soluble toxin that causes death of fish and other marine animals. Several methods were tested to visualize plate morphology of Cryptoperidiniopsis brodyi and Pfiesteria piscicida. Cellulose plates of cells were exposed and visualized- by a membrane stripping method using Triton X-100. While calcofluor M2R white stain could readily bind to the thecal plates, details of the plate tabulation were difficult to observe. Fixation with osmium tetroxide $(OsO_4)$ produced well preserved cells with little morphological distortion, but thecal plates could not be visualized. Scanning electron microscopy (SEM) observation using the membrane stripping method showed distinctive plate tabulations between C. brodyi and P. piscicida suggesting that this method is a useful tool for morphological identification of lightly armored dinoflagellates.

• 한국의류학회지
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• 제10권3호
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• pp.1-8
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• 1986

The purpose of this study was to investigate the characteristics of human epidermal stratum corneum as protein model soils for detergency test. The stratum corneum was collected by scraping of the skin and purified with solvent. The results obtained were as follows: 1. Purified stratum corneum contained $92.38\%$ of crude protein. 2. In the amino acid compositions, contents of glycine, glutamic acid and serine were high and methionine and cystine were low. They were similar to fibrous $\alpha$-keratin consisted of stratum corneum. Whereas the content of polar amino acids was decreased, that of nonpolar amino acids was increased after enzyme hydrolysis. 3. The hydrolysis of stratum corneum with enzyme increased muck at initial reaction time and levelled off in 4$\~$6 hours. The hydrolysis with enzyme was improved effectively at its optimum temperature and pH. 4. The hydrolysis of stratum corneum with enzyme increased by the addition of surfactants. The order of compatibility with enzyme was in the order of Triton X-100>AOS>LAS.

• Journal of Preventive Medicine and Public Health
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• 제19권1호
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• pp.76-84
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• 1986

Analyses of lead and zinc were made by means of standard addition method using atomic absorption spectrophotometer(Baird Ltd., Model A5100) with flameless method for lead and flame method for zinc. The blood samples used were merely diluted with triton x-100, because it was simple, rapid and minimal risk of contamination. Mean recovery rate for lead added to the blood ranged from 97.7 to 101.3% with coefficient of variation ranging from 1.9 to 10.7%, and that for the added zinc ranged from 99.0 to 102.2% with coefficient of variation ranging from 2.1 to 9.1%. In repeated measurements of zinc in the blood, good reproducibility and interindividual variation were proved(p<0.01). In comparison of the lead and zinc concentrations in the blood determined by the standard addition method and standard method, there were good correlations between 2 sets of data (r=0.9731 for lead and r=0.9785 for zinc), although lead levels were estimated higher by the former method(p<0.01) and zinc levels by the latter method(p<0.01). It can be concluded that lead and zinc levels in blood standard addition method is reliable for determination of lead and zinc in the blood with good accuracy and reproducibility.

• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2003년도 International Symposium on Clean Environment
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• pp.103-108
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• 2003

A method to determine chromium(III) ion in aqueous solution by chemiluminescence method using a lucigenin entrapped silica sol-gel film has been studied. An optical probe for chromium(III) ion has been prepared by entrapping lucigenin into silica sol-gel film coated on a glass support by dip coating. The chromium(III) optical sensor is based on the catalytic effect of chromium(IIII) ion on the reaction between lucigenin and hydrogen peroxide in basic solutions. The effects of Nafion, DMF and Triton X-100 were investigated to find the optimum condition to minimize cracking and leaching from the probe. The effects of pH and concentrations of lucigenin and hydrogen peroxide on the chemiluminescence intensity were investigated. The chemiluminescence intensity was increased linearly with increasing chromium(III) concentration from $2.5{\times}10^{-4}$M to $8.0{\times}10^{-7}$M and the detection limit was $4.0{\times}10^{-7}$M.

• BMB Reports
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• 제29권5호
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• pp.436-441
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• 1996

The palmitoyl acyl carrier protein (ACP) specific thioesterase (EC 3.1.2.14) from Iris pseudoacorus was purified and characterized. The thioesterase which was very unstable in relatively high salt concentrations was eluted using a co-gradient of Triton X-100 and low concentration of KCl or Na-phosphate from Q-Sepharose, DEAE-Sepharose, and hydroxyapatite chromatography. SDS-PAGE analysis showed a single band with a molecular weight of 35,000. The native molecular weight of approximately 37,000 was estimated by Sephacryl S-200 chromatography, indicating that the enzyme is a monomer. The thioesterase activity was inhibited about 75% and 50% by N-ethylmaleimide (2 mM) and phenylmethylsulfonyl fluoride (2 mM). respectively. The N-ethylmaleimide-inactivation was protected by sodium palmitate but the inactivation with phenylmethylsulfonyl fluoride was not protected. Oxidation of thiols by 2 mM 5.5'-dithio-bis-(2-nitrobenzoic acid) resulted in 65% inactivation of the enzyme. These results suggest that a cysteinyl residue is essential to the catalytic reaction of the enzyme. The enzyme activity was increased by sodium citrate and also by $Cu^{2+}$

• 한국환경농학회지
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• 제36권2호
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• pp.135-139
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• 2017

BACKGROUND: Identification and characterization of endophytic yeasts inhabiting the roots of Ulmus parvifolia Jacq. and Quercus salicina Blume require biotechnological and culture-based techniques. METHODS AND RESULTS: Homogenized U. parvifolia and Q. salicina root samples were spread onto four types of agar medium containing ancgtibiotics, L-sorbose, and Triton X-100. In total, 25 yeast strains were isolated and subjected to phylogenetic analysis based on their internal transcribed spacer region sequences. The results revealed that the yeast genera Cyberlindnera (12 isolates) and Cryptococcus (1 isolate) were associated with roots of U. parvifolia; and the genera Rhodotorula (8 isolates), Trichosporon (3 isolates), and Kluyveromyces (1 isolate) were associated with roots of Q. salicina. Additionally, a Kluyveromyces isolate produced a detectable level of bioethanol. The yeast strains reported herein may be used in industrial production of biosurfactants and bioethanol. CONCLUSION: Our findings revealed that the endophytic yeast genera Cyberlindnera and Cryptococcus predominated in roots of U. parvifolia; and the genera Rhodotorula (8 isolates), Trichosporon (3 isolates), and Kluyveromyces (1 isolate) predominated in roots of Q. salicina. Additionally, Kluyveromyces isolates produced a detectable level of bioethanol.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.145-146
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• 2000

The presence of cross-reacting muramidase in Lactobacillus delbrueckii ssp. bulgaricus ULl2 was shown by using monoclonal antibodies raised against an muramidase-2 of Enterococcus hirae ATCC 9790. The separation of protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western immunoblot confirmed the presence of one cross-reacting band in Enterococcus hirae with an estimated molecular mass of 80 kDa, L. bulgaricus cultured cells harvested after 4 and 12 h were submitted to different autolysin releasing procedures and the liberated products were allowed to cross-react with muramidase-2 antibodies in order to estimate the efficiency of each treatment. Although the cultured cells harvested after 4 h yielded only a slight immune-reaction in Western immunoblots against these enzyme monoclonal antibodies, a strong signal was observed for the cell walls obtained from the same experimental conditions and treated with Triton X-100 surfactant. The same phenomenon was also observed by light fluorescence microscopy. Immune-labelling followed by optical and electron microscopy have shown that the muramidase-2 of L. bulgaricus ULl2 was essentially localized in the innermost part of the cell wall.(omitted)