• KSBB Journal
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• v.10 no.1
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• pp.30-37
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• 1995

Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32$^{\circ}C$, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.

• KSBB Journal
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• v.15 no.3
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• pp.318-322
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• 2000

An oxygen-sensitive fumarate reductase has been purified from the cytosol fraction of the Enterococcus faecalis RKY1 grown anaerobically on a defined medium containing glycerol and fumarate. A major portion of the purification was performed with employing Triton X-100 and reducing agents by Phenyl-sepharose CL-4B DEAE-sepharose and Dephadex G-150 The final activity was 0.42 unit/mg. The deduced molecular mass of active band was 66 kDa. The optimal pH and temperature for the activity were 7.0 and 38$^{\circ}C$ respectively. The enzyme activity was not affected by 1mM metal ions such as bacl2 $.$2H2O HgCl2 MnCl2$.$4H2O ZnCl2 CuCl2$.$2H2O Mgcl2$.$6H2O FeSo4$.$7H2O and by EDTA. Partially purified enzyme ws yellow in color ; spectroscopic study indicated the presence of flavins as a cofactor.

• The Korean Journal of Food And Nutrition
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• v.9 no.3
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• pp.253-258
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• 1996

Stabilities of sweets potato f-amylase on various reagents were studied. The enzyme was stabilized by bovine serum albumin, Triton X-100 and 2-mercaptoethanol of 0.04%. Among them, bovine serum albumin was the most effective. And enzyme stability was increased by using the deairated solution. The enzyme activity was remained 0% in the absence of glycerol, 25% in the presence of 20% glycerol and 50% in the presence of 40% glycerol at 37$^{\circ}C$, for 15 hours in pH 11. SDS inhibited the enzyme, and 2-mercaptoethanol and dithiothreitol stabilized it.

• Biomedical Science Letters
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• v.18 no.2
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• pp.96-103
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• 2012

Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.

• Mycobiology
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• v.43 no.2
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• pp.157-162
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• 2015

Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.

• Journal of Environmental Science International
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• v.6 no.5
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• pp.481-488
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• 1997

Microorganisms capable of degrading trlchloroethylene(TCEI using phenol as a induction substrate were isolated from industrial effluents and soil. The strain MS-64K which had the highest blodegradablllty was identified as the genus Micrococcus. The optimal conditions of medium for the growth and blodegadatlon of trlchloroethylene were observed as follows; the initial pH 7.0, trlchloroethylene 1, 000ppm as the carbon source, 0.2% ${(NH_4)}_2SO_4$, as the nitrogen source. respectively. Lag period and degradation time on optimal medium were shorter than those on Isolation medium. Growth on the optimal medium was Increased. Addition of 0.1% Triton X-100 Increased the growth rate of Micrococcus sp. MS-64K, but degradation was equal to optimal medium. Trlchloroethylene degradation by Micrococcus sp. MS-64K was shown to fit logarithmic model when the compound was added at initial concentration of 1, 000ppm.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.91-91
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• 2002

There are many methods to introduce exogenous DNA into embryo for the purpose of producing transgenic animals. Exogenous gene can be integrated into oocyte as a form of sperm vector. In this study, sperm was used as a vector for transgene that is enhanced green fluorescent protein (EGFP). The objective of this study was to investigate the expression of exogenous gene in bovine embryos after injection of spermatozoa cocultured with EGFP fragment. Spermatozoa were plunged into liquid nitrogen and thawed several times or shaked in 0.2% Triton X-100 to remove sperm membrane which followed by DTT treatment. (omitted)

• Korean Journal of Environmental Agriculture
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• v.35 no.2
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• pp.137-142
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• 2016

BACKGROUND: Identification of endophytic yeasts inhabiting the internal roots of the Mankyua chejuense tree requires techniques involving biotechnology. There is a need for a culture-based method to isolate and identify yeast strains associated with M. chejuense.METHODS AND RESULTS: We spread homogenized M. chejuense root samples onto glucose-peptone- yeast agar containing antibiotics, Triton X-100, and L-sorbose. A total of 152 yeast isolates were obtained and identified via phylogenetic analysis based on ITS gene sequencing. The results revealed that the root-associated yeast species included the genera Cyberlindnera (140 isolates), Candida (11 isolates), and Kluyveromyces (one isolate). Additionally, three yeast isolates showed high bioethanol production.CONCLUSION: We identified the specific yeast community associated with M. chejuense roots. These yeast isolates may have industrial applications as bioethanol producers. Our findings revealed that Cyberlindnera isolates included C. suaverolens and C. satumus, while Kluyveromyces isolates showed high bioethanol production.

• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.378-385
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• 1997

Cation motive ATPases on cell membranes of Helicobacter pylori were investigated using everted membrane vesicles. Latent ATPases could be ascertained from aggregated vesicle using N, N-dimethylformamide (DMF) and Triton X-100. By contrast, ultrasonication or chloroform treatments caused membranes to be disrupted, resulting in an alteration of sensitivities against azide or vanadate. Considerable amounts of vanadate-sensitive enzymes were identified from vesicle micelles, prepared by the dilution method. These were activated in the presence of either $Ni^{2+}\;or\;NH_4^+$. From studies employing H. pylori intact cell systems, we found that ATPase expression of this bacterium was markedly dependent upon air composition. It was interesting that cellular expression of $Ni^{2+}$- or $NH_4^{+}$-motive ATPases was significantly affected by extracellular pH, suggesting that these unique enzymes may physiologically be involved in cellular $Ni^2$ import and $NH_4^+$ export, respectively.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1998.11a
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• pp.174-177
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• 1998

The effects of surfactants affecting polycyclic Aromatic Hydrocarbon(PAHs) solubility and biodegradation in soil slurry were investigated. The critical micelle concentration(CMC) values of surfactants used in this study were 12.7mg/L(Brij 30), 13.4mg/L(Tween 80), 13.6mg/t(Triton X-100). The solubility of PAH increased as the Hydrophile-Lipophile Balance(HLB) value of surfactant decrease. At surfactant biodegradation and toxicity experiement using respirometer, Brij 30 did not show any toxic effect and substrate inhibition upon the level of 1.5g/L. Also, biodegradation of Brij 30 gave no reduction on the phenanthrene biodegradation rate. When the desorption rate of phenanthrene between sand and clay is compared, lower percentage of phenanthrene was desorbed at clay because of the larger surface aera and higher organic content of clay. At the biodegradation experiments of phenanthrene in soil slurry phase, more than 90% of initial phenanthrene adsorbed onto both sand and clay were biodegraded by phenanthrene- acclimated cultures.