• Journal of Korean Society of Forest Science
• /
• v.35 no.1
• /
• pp.24-32
• /
• 1977

According to the chlorous salt method, most of holocellulose whose lignin was removed, was obtained. In extracting xylan from holocellulose by the different densities of alkali, 5% KOH was extracted three times but still there remained part of xylan in it and another composite of hemicellulose and cellulose was obtained. The extraction of 10% and 20% KOH showed a desirable result. Rather than the ordinary method to use a large quantity of ethanol in the precipitation isolation of xylan, the method to use a small quantity of ethanol in adopting the dialysis with cellophane-membrane by condensing density to one tenth, made il possible to extract a high purity xylan in a high retrieving rate. In isolating glucomannan, the residue of 5% KOH extraction contained a large quantity of xylan, the residue of 10% and 24% KOH extraction, also showed the same result and the comparison between glucose and mannose was approximately 1 : 1. The purification of Fehling solution made it possible to obtain comparatively pure xylan but the process of oxidation dissolution was complicated and the retrieving rate was low. This was not a good method. The ethanol titration purified a high purity xylan in a high retrieving rate and was a very excellent purifying method, considering its simple and easy process. These two purifying methods, however, could not completely remove the residue of arabinose. This will be examined and reported later.

• YAKHAK HOEJI
• /
• v.30 no.1
• /
• pp.31-41
• /
• 1986

Sarcolemmal membrane fraction from canine ventricle was isolated from the discarded pellet after the first homogenization in the isolation procedure of sarcoplasmic reticulum (Method 1) and the protein yield, purity, and sidedness of this preparation were compared to those of sarcolemmal fraction prepared by method of Lee et al. (Method 2) and a slight modification of original protocol of Jones et al. (Method 3). Method 1 differed from Method 2 essentially only in that vigorous homogenization was carried out by omnimixer and homogenization medium containing 30mM Tris-maleate was used in the first step. The sarcolemmal fraction was enriched from 45 to 50 and 29-fold in [$^3H$] ouabain, [$^3H$] DHA, [$^3H$] QNB binding and $Na^+$, $K^+$-ATPase activity, respectively, compared to homogenate. Total $Na^+$, $K^+$-ATPase activity of highly sarcolemma enriched fraction was 144.6$\pm$16.4$\mu\textrm{mol}$ Pi/mg protein/hr, which was about 85%, of total ATPase activity, and the yield of the preparation was 15.7 mg protein per 100g of starting ventricular tissue. The sarcolemmal preparation supported $^{45}Ca^{2+}$-uptake in the presence of ATP but this uptake was not dependent on oxalate. Sarcolemmal $Na^+$, $K^+$-ATPase activity and detectable [$^3H$] ouabain binding were increased about 32% and 35%, respectively, by pretreatment of sarcolemmal fraction with optimal concentration of sodium dodecylsulfate (0.3-0.4mg/mg protein), suggesting that this preparation contained about 24% of sealed rightside-out vesicles, 26% of sealed inside-out vesicles, and 5001o of freely permeable (leaky) form. This procedure showed the highest protein yield and leaky population, compared to Method 2 and 3. On the other hand, sarcolemmal fraction prepared by Method 2 and 3 showed low value in protein yield but comtained high population of inside-out (46%) and rightside-out (49%) vesicles, respectively, compared to present procedure (Method 1). The results indicate that vigorous homogenization decreases the population of sealed sarcolemmal vesicles but increases the sarcolemmal protein yield per gram tissue and that this procedure is available for further purification of sarcolemmal fraction and for the receptor binding study of sarcolemma.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.51 no.spc1
• /
• pp.166-173
• /
• 2006

Soyasaponins are phytochemicals of major interest fur their health benefits. Chemical investigation of a soybean phytochemical concentrate resulted in the isolation and identification of triterpenoid saponins. The MeOH extraction of defatted hypocotyl separated from soybeans was peformed by the automated solvent extractor (ASE). Fractionation was performed on a flash column ($150mm{\times}40mm$ i.d.) packed with a preparative $C_{18}$ reverse phase bulk packing material $(125\AA,\;55-105{\mu}m)$ and monitored at 210 nm, and collected 14 fractions. Consequent Fsat preparative column liquid chromatography (Fast PCLC) was performed for the purification of Fraction-I (Fr-I) collected from the fraction 8 and 9 of flash chromatography. Fsat PCLC was performed on a Luna $C_{18}\;10{\mu}m,\;100{\AA}$, semipreparative reverse phase column ($250cm{\times}50mm$ i.d.) for the purification of isolated unknown compound (Fr-I-2). Chemical structure of acetyl soyasaponin $A_1\;(MW:1436.6,\;C_{67}H_{104}O_{33})$ was identified and determined by a combination of extensive NMR ($^1H-NMR$, 400 MHz; $^{13}C-NMR$, 100 MHz; DEPT), IR, UV, and ESI-MS analysis.

• Journal of the Korean Wood Science and Technology
• /
• v.28 no.3
• /
• pp.25-33
• /
• 2000

This study was carried out to find the optimal isolation conditions of xylan from steam-exploded materials, such as rice straw(Oryza sativa), barley straw(Hordeum vulgare) and oak wood(Quercus mongolica), In the chemical composition, we found that the contents of water-extractives and ash of rice straw and barley straw were more than those of oak wood. Rice straw, barley straw and oak wood were steam-exploded at 20kgf/$cm^2$ for 3 minutes or 6 minutes. The content of lignin in three different steam-exploded materials was higher than that of non-treated materials. The crude xylan was extracted with hot water and 0.5% KOH solution from steam-exploded materials. In the sugar type of crude xylan extracted with hot water and 0.5% KOH solution, the oligomer content of crude xylan extracted with hot water was much more than that of crude xylan extracted with 0.5% KOH solution. The crude xylan was purified with 5% barium hydroxide Solution and ethanol precipitation procedure. The content of xylose of purified xylan was over 85%, but other sugar residues(arabinose, mannose, galactose and glucose) were not removed completely.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.10
• /
• pp.1021-1025
• /
• 2011

A Bacillus subtilis strain was isolated from the intestine of Sebastiscus marmoratus (scorpion fish) that was identified as Bacillus subtilis CH2 by morphological, biochemical, and genetic analyses. The chitosanase of Bacillus subtilis CH2 was best induced by fructose and not induced with chitosan, unlike other chitosanases. The strain was incubated in LB broth, and the chitosanase secreted into the medium was concentrated with ammonium sulfate precipitation and purified by gel permeation chromatography. The molecular mass of the purified chitosanase was detected as 29 kDa. The optimum pH and temperature of the purified chitosanase were 5.5 and $60^{\circ}C$, respectively. The purified chitosanase was continuously thermostable at $40^{\circ}C$. The specific acitivity of the purified chitosanase was 161 units/mg. The N-terminal amino acid sequence was analyzed for future study.

• Food Science of Animal Resources
• /
• v.36 no.6
• /
• pp.791-798
• /
• 2016

Free radicals may attack cells or tissue, leading to chronic diseases, and antioxidant consumption is potentially useful for removing free radicals. Egg proteins may be used as potential sources of antioxidant considering their ability of scavenging free radicals to apply for food or cosmetics industry. In this study, we obtained a natural antioxidant protein from fertilized eggs, which was a dietary supplement in some Asian countries. Meanwhile, antioxidant activities of these proteins were evaluated using different oxidation systems. With increasing incubation time, the antioxidant activity of these proteins increased during 15 d of incubation. The samples on day 15 were performed for isolation of antioxidant protein. The protein, named P4-1 (MW, 45 kDa), was isolated and purified by consecutive chromatographic methods. P4-1 contained 17 amino acids, which was determined by liquid chromatography-mass spectrometry and Amino Acid Analyzer. Moreover, the amino acid sequence was highly similar to that of ovalbumin. Differential scanning calorimetry showed that the denaturation temperature of P4-1 was $57.16^{\circ}C$. Furthermore, P4-1 suggested high oxygen radical-absorbance activity in ${\cdot}OH$ assays, and its antioxidant activity was stable at $30-50^{\circ}C$ in acidic and neutral pH. Thus, these results revealed that P4-1 may be a potential resource as a natural antioxidant.

• Journal of the Korean Chemical Society
• /
• v.38 no.8
• /
• pp.608-615
• /
• 1994

The isolation, purification and characterization of a carotenoprotein from the carapace of Pnaeus orientalis were investigated. The carotenoprotein was purple with broad λmax between 480, 409, 318 and 280 nm. Apparent structures were estimated by using X-ray diffractometry and scanning electron microscope, respectively. The molecular weight of the carotenoprotein complex had been determined by GPC and PAGE. The heavier complex, designated the $\alpha$-form (M.W = 170 KDa), was dissociated to a major subunit, $\beta$-form (M.W = 42 KDa). SDS-PAGE of $\alpha$-form showed apparently oligomeric pattern, and also $\beta$-form gave two polypeptides corresponding to 22 KDa and 19 KDa, respectively. The amino acid of the two proteins $({\alpha}-and\;{\beta})$-form, lipid and free fatty acid compositions were described. The prosthetic groups of the carotenoprotein were confirmed by TLC, IR, $^1H$-NMR, MS and various organic reactions as astaxanthin, astaxanthin monoester and astaxnathin diester.

• Journal of Biomedical Engineering Research
• /
• v.44 no.6
• /
• pp.465-475
• /
• 2023

In this study, we demonstrated a silica nanofibrous membrane based on the electrospinning process and evaluated its DNA isolation and purification performance in PCR pretreatment. Generally, silica membranes made of non-woven fabric are used for PCR pretreatment, but this study aimed to improve the efficiency of the pretreatment process by developing a nanofiber-type silica membrane with high specific surface area and porosity. In order to manufacture a nanofiber-shaped silica film while maintaining the original physical properties of silica, nanofiber membranes produced by adding various concentrations of PEO (5 wt%, 8 wt%, and 10 wt%) to silica prepared by the sol-gel method were compared. In terms of nanofiber membrane production, the higher the PEO concentration, the more effective it was in producing nanofiber membranes. The produced silica nanofiber membrane was inserted to a pretreatment device used in commercial PCR equipment, and the pretreatment performance was compared and verified using Salmonella bacteria. When Salmonella was used, samples containing 5 wt% PEO showed superior PCR efficiency compared to samples containing 8 wt% and 10 wt% PEO. These results show that adding 5 wt% of PEO can effectively improve DNA purification and separation by producing a nanofiber-shaped silica film while maintaining the physical properties of silica. We expect that this study will contribute to the development of effective PCR pretreatment technology essential for various molecular biology applications.

• Journal of Life Science
• /
• v.20 no.12
• /
• pp.1801-1806
• /
• 2010

A $\beta$-xylosidase encoding gene from Klebsiella sp. Sc was cloned in Escherichia coli. The $\beta$-xylosidase gene consisted of an open reading frame of 1,680 nucleotides and encodes 559 amino acids with a deduced molecular weight of 63 kDa. The deduced amino acid sequence of the $\beta$-xylosidase from Klebsiella sp. Sc exhibits 90% identities and 95% positives compared to those from Klebsiella oxytoca (KOX), Lactobacillus lactis (LAC, 82%, 90%), Bacillus longum (BLON, 69%, 81%) and Escherichia coli (ECOLI, 47%, 63%). The $\beta$-xylosidase was purified by GST-fusion purification system. The pH and temperature optima of the enzyme were 6.6 and $55^{\circ}C$, respectively. The $\beta$-xylosidase hydrolyzes xylobiose to xylose.

• KSBB Journal
• /
• v.14 no.6
• /
• pp.677-681
• /
• 1999

Purificationi of egg yolk immunoglobulin(IgY) was performed to understand the property of egg immunoglobulin. IgY differs from mammalian IgY in the molecular size(larger), isoelectric point(more acidic), and binding ability with mammalian complement and protein A(nonbinding ability). IgY is also known as ${\gamma}$-livetin and exists in egg yolk together with other two water-solubel proteins, ${\alpha}$-livetin(chicken serum albumin) and ${\beta}$-livetin(${\alpha}_2$-glycoprotein) and various lipoproteins(Low density lipoprotein, LDL and High density lipoprotein, HDL) which are the major components of egg yolk. The first step of isolation of IgY is to separate the water-solube proteins from lipoproteins. We report a simple method for separation of water soluble proteins using k-carrageenan and sedimentation. k-carrageenan was found to be effective for removal of yolk lipoprotein as a precipitate. IgY remained supernatant, and was isolated by chromatography on columns of DEAE-Sephacel and G 75 gel filtration chromatography.