• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1516-1524
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• 2014

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.44-50
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• 1997

$\beta $-Glucosidase (EC 3.2.1.21) was purified from Aspergillus niger SFN-416 by a sequential process of ammonium sulfate precipitation, Sepadex G-100 and DEAE-Sephacel column chromatography. Molecular weight of the enzyme was 46, 000 daltons. The K$_{m}$ and V$_{max}$ values for PNPG were 0.67 mM and 25 moles/ml $\cdot $min., respectively. The optimum pH and temperature of the enzyme activity were 3.5 and 58$\circ $C, respectively. The enzyme activity was decreased by addition of metal ions, and increased by addition of metanol, ethanol, isopropanol and 1-butanol at a concentration of 10% (v/v). Stability of the enzyme was increased by addition of isopropanol and 1-butanol at a concentration of 10% (v/v).

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.6
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• pp.1166-1172
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• 1998

A method for the isolation and purification of DNA from a red algae, Porphyra was innovated. The innovation of the method consists mainly of three steps that include sodium acetate treatment, chloroform extraction, and 0.2 volume isopropanol precipitation step. The sodium acetate treatment was designed to remove polysaccharide contamination, and the isopropanol step to remove proteins and salts contaminents. Genomic DNA,s of several species(for example, P. tenera, P. yezoensis, P. seriata, and P. pseudolinearis) was successfully isolated by the innovated method. The amount of DNA purified from one g of sample material with the innovated method was 53 g in average. The resulting DNA was characterized to include high molecular weight and showed no nuclease activity. The DNA was pure enough to be digested directly by various restriction enzymes without any difficulties. Porphyra DNA was pure enough and adequate for amplification reaction through the polymerase chain reaction (small nuclear rDNA PCR amplification).

• Food Science and Biotechnology
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• v.18 no.5
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• pp.1150-1154
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• 2009

Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RT-PCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.

• Membrane Journal
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• v.6 no.3
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• pp.157-165
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• 1996

Polysulfone(PS) membranes were prepared from homogeneous PS solutions by the phase inversion technique. When propionic acid(PA) was added into a casting solution of n-methylpyrrolidone(NMP) and PS, precipitation rate of the solution film was accelerated. This kind of acceleration was consistent, even though a precipitating nonsolvent was changed from water to isopropanol. These phenomena were caused by decrease of nonsolvent tolerance in the casting solution due to addition of PA. PS powder was prepared by precipitation of a 3wt% solution in dimethylformamide(DMF) using ethanol as nonsolvent. Gas adsorption analysis of the powder showed that the capillary condensation sites were found in the powder structure. Membranes prepared from PS solution(15wt%) in NMP had the following characteristics of gas adsorption and water permeation. In gas adsorption analysis, the membrane precipitated using isopropanol showed low uptake of nitrogen gas and the capillary condensation sites were not found. On the contrary, a significant amount of the capillary condensation sites was found in the membrane coagulated by water, which was related to increase of nitrogen uptake. tn the membrane prepared froin the solution including PA, an increase of the Henry's law sites and the Langmuir sites was not found clearly. However, the capillary condensation sites were significantly increased, and the water transport also increased.

• Journal of the Korean Ceramic Society
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• v.41 no.10 s.269
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• pp.742-748
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• 2004

Isopropanol of low boiling point was used as a solvent to prepare Al-doped ZnO(AZO) thin films. A homogeneous and stable sol was made from Zn acetate a solute whose mole concentration was 0.7mol/$\iota$ and Al chloride as a dopant. Al-doped ZnO thin films were prepared by sol-gel method as a function of post-heating temperature from 500 to $700^{\circ}C$ and the optical and electrical properties were investigated. The c-axis orientation along (002) plane was enhanced with the increasing of post-heating temperature and the surface morphology of the films showed a homogeneous and nano-sized microstructure. The optical transmittance of the films post-heated below $650^{\circ}C$ was over $86\%$, but decreased at $700^{\circ}C$. The electrical resistivity of the thin films decreased from 73 to 22 $\Omega$-cm as the post-heating temperature increased up to $650^{\circ}C$, but increased greatly to 580 $\Omega$-cm at $700^{\circ}C$. XPS analysis indicated that the deterioration of electrical and optical properties was attributed to the precipitation of $Al_2O_3$ phase on the surface of AZO thin film. This result suggests that the optimum post-heating temperature to improve electrical and optical properties is $600^{\circ}C$.

• Applied Chemistry for Engineering
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• v.19 no.6
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• pp.633-639
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• 2008

Polysulfone membranes were prepared via the phase inversion process. Toluene was added as a nonsolvent additive in the casting solution containing a mixture of polysulfone and n-methylpyrrolidone. When prepared via the diffusion-induced process using isopropanol as a precipitation nonsolvent, the solidified membranes revealed a similar asymmetric structure irrespective of the addition of toluene, presenting both a dense skin layer and a sponge-like support layer. The added toluene played a role of enhancing liquid-liquid phase separation of the casting solution, and skin layer thickness of a prepared membrane increased with toluene content in the casting solution. On membrane performance, the solute rejection showed a uniform behavior irrespective of the addition of toluene. However, in spite of the significant increase in dense skin layer thickness, the water permeation through the membrane prepared with 60 wt% toluene revealed five times as much flux, compared with that of the membrane prepared without toluene additive.

• ALGAE
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• v.21 no.2
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• pp.261-266
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• 2006

The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.5
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• pp.821-826
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• 1998

Functional properties of mucilage and pigment extracted from Opuntia ficus-indica var. saboten were determined at various temperatures, pHs and alcohol concentrations. The crude mucilage extracted from pickly pear showed pH 4.2, 0.14% total acidity and 8.1% total soluble solid content(w/w, wet basis). Polysaccharide was purified from mucilage extract by isopropanol precipitation. Intrinsic viscosity of polysaccharide was 18.1dl/g and decreased with increasing KCl concentration. Relative viscosity and color stability of mucilage extract were determined with capillary viscometer and spectrophotometer at 534nm, respectively. In additions of 1~20%(v/v) ethanol, the red pigment of mucilage extract was very stable, but relative viscosity, increased gradually. For heating above 7$0^{\circ}C$, the stability of red pigment decreased drastically, but rheological property of mucilage was not changed. During storage, the red pigment was extremely unstable at above pH 8.3. At both pH 3.0 and pH 4.2, the red pigment was the most stable at 4$^{\circ}C$ for 18 days. In the case of storage at 37$^{\circ}C$, pigment of mucilage extract at pH 3.0 was destroyed more quickly than that at pH 4.2. Natural mucilage extract(pH 4.2) showed the good stability of red pigment at 3$0^{\circ}C$ for 10 days.

• Korean Journal of Food Science and Technology
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• v.9 no.4
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• pp.251-254
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• 1977

Pectin, hesperidin and naringin were extracted from hot air-dried peel ($60^{\circ}C$, 1 hr and air velocity 160 fpm) of citrus produced in Korea in order to see the amount of each component contained in the peel. Pectin was extracted by three different methods and the quality and contents of the pectins were determined respectively. 1. The pectin yield by the total pectic substance method was the highest (26.0% for unshiu (U) and 28.5% for natsudaidai (N) expectedly and the soluble pectic substance method the least (13.5%(U) and 15.6% (N)) The yield by method III (extraction by water at pH 1.5 followed by isopropanol precipitation) was intermediate (18.1% (U) and 20.8%(N)). Anhydrouronic acid (AUA) content was the highest (92.0% (U) and 90.3%(N)) in those by method III. The AUA contents of the other pectins were 80.0% for soluble pectin (for both U and N), 71.6% for the commercial pectin (Sunkist Groups Inc., U.S.A.), 58.0%(N) and 63.4%(U) for total pectic substance. 2. The methoxyl content of total pectic substance was the lowest (4.81%(U) and 4.88%(N)). However, there was no significant difference in methoxyl content among the rest which were found to have low levels(5.27-7.20%). 3. The pectin by method III gave the highest jelly strength. The commercial pectin, soluble pectice substance and total pectic substance were next in order. 4. The hesperidin content of unshiu was 5.07% (dry basis) and the naringin content of natsudaidai 3.03% (dry basis).