• Journal of Life Science
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• v.24 no.6
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• pp.603-611
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• 2014

The lectins were separated from Korean and Chinese mung bean seeds finally via chromatography using Sephadex G-100 and their biochemical features were studied and compared. They showed no hemagglutination with human red blood cells regardless of trypsin treatment and showed hemagglutination with only trypsin treated rabbit red blood cells. The molecular weights of two lectins were identified as 54 kDa and 28 kDa by SDS-PAGE. It was found that while the optimal reaction temperature of the lectin from Korean mung bean was $60^{\circ}C$, that of the lectin from Chinese mung bean seeds was $50^{\circ}C$. It was found also that the most thermal stable temperature of the seed lectin from Korean mung bean seeds was $50^{\circ}C$ and the lectin from Chinese mung bean was $40-50^{\circ}C$. The lectin from Korean mung bean seeds showed the highest activity at pH 3.2 and the lectin from Chinese mung bean showed the highest activity at pH 6.2. It was identified that when treating a denaturant, thiourea and guanidine-HCl resulted in no hemagglutination, so they induced denaturalization. It was identified also that there was no hemagglutination with urea, so it did not induced denaturalization. They showed no septicity to 6 types of carbohydrates including D-glucose. In addition, the lectins from the two mung bean seed had specificity to metal ions.

• KSBB Journal
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• v.25 no.5
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• pp.429-436
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• 2010

Keratinase is widely used in certain industrial applications. The present study sought to improve the culture conditions of Bacillus subtilis SMMJ-2 to facilitate mass production of keratinase. Strain SMMJ-2 was irradiated by ultraviolet light and the resulting isolates were tested for keratinase activity. Isolates displaying elevated keratinase activity were selected and used to determine the optimum temperature (24, 30, 37, 45, $55^{\circ}C$) for bacterial keratinase production during a 4 day incubation period. The highest enzyme activity (55 units/mL/min), from a Bacillus subtilis SMMJ-2 mutant (mutant No. 2) was demonstrated following incubation at $30^{\circ}C$. The effects of carbon and nitrogen sources on keratinase production were confirmed by measuring the enzyme activity from the culture broth of the mutant strain cultured in various media containing different carbon source and nitrogen sources during a 4 day period. The optimal medium composition for producing keratinase consisted of 1% glucose, 0.7% $K_2HPO_4$, 0.2% $K_2HPO_4$, and 1.2% soybean meal. Optimal initial pH and temperature for producing keratinase were 7.0 and $30^{\circ}C$, respectively. Keratinases produced by B. subtilis SMMJ-2 and the mutant No. 2 were purified from the culture broth which used soybean meal as a nitrogen source. Membrane ultrafiltration, DEAE-sephacel ion exchange and Sephadex G-100 gel chromatography were used to purify the enzymes. The purified keratinases from both B. subtilis SMMJ-2 and the mutant No. 2 showed single bands and their molecular weights were estimated as 28 kDa and 42 kDa, respectively on SDS-polyacrylamide gel electrophoresis.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.198-206
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• 2010

The aim of this work was to investigate the synergically bactericidal effects and cellular responses of tea polyphenols (TPP) and imipenem on imipenem-resistant Pseudomonas aeruginosa. Imipenem-resistant Ps. aeruginosa was isolated from patient in hospital. The bactericidal effects of TPP and imipenem were evaluated on the basis of its minimum inhibitory concentrations (MIC). The combined use of TPP and imipenem resulted in 16-fold and 8-fold reductions in the MICs of imipenem for the imipenem-susceptible and imipenem-resistant Ps. aeruginosa, respectively. The bactericidal effects of the imipenem and TPP against the Ps. aeruginosa was evaluated using the time-kill assay. The synergetic effects of the combinations of TPP and imipenem against Ps. aeruginosa were confirmed. Western blot using anti-DnaK and anti-GroEL monoclonal antibodies was performed to investigate the expression of stress shock proteins (SSPs) in imipenem-susceptible and imipenem-resistant strains exposed to TPP. The amount of SSPs were induced as the exposure time increased and decreased. The molecular weights of DnaK and GroEL were 70 kDa and 60 kDa, respectively. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides (LPS) increased or decreased in the strain treated to different concentrations and exposing periods of TPP. Scanning electron microscopic analysis demonstrated the presence of umblicated and wrinkled surfaces for cells treated with TPP or imipenem.

• Applied Biological Chemistry
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• v.40 no.5
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• pp.375-379
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• 1997

Two ribosome-inactivating proteins, PRIP 1 and PRIP 2 have been isolated from the leaves of $Cucurbita\;moschata\;D_{UCHESNE}$. Crude extracts were purified through ammonium sulfate precipitation and column chromatography using DE-52 cellulose, S-Sepharose, FPLC Suprose 12 HR and FPLC Mono-S. The molecular weights of PRIP 1 and PRIP 2 were 31,000 and 30,500, respectively. PRIP 2 was thermostabe and maintained its activity even after the incubation of the protein at $50^{\circ}C$ for 30 min. In a cell free in vitro translation system using rabbit reticulocyte lysate, protein synthesis was inhibited by the addition of PRIP 1 and PRIP 2. The $IC_{50}$ of PRIP 1 and PRIP 2 were 0.82 nM and 0.79 nM, respectively. The comparison of N-terminal amino acid sequences of the PRIP 1 and PRIP 2 with known RIPs revealed that PRIP 1 shows sequence similarity with Luffin B from Luffa cylindrica and Trichokirin from Trichosanthes kirilowii Maximowicz and PRH) 2 has sequence similarity with Momordin II and MAP 30 from Momordica charantia.

• Applied Biological Chemistry
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• v.49 no.3
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• pp.215-220
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• 2006

Response surface methodology was implemented to determine an optimal extraction condition in Phellinus linteus water extract. Extraction was performed on 10 experimental conditions including independent variables such as extraction time $(1{\sim}5\;hrs)$ and water volume over sample (sample : $H_2O$ = 1 : $40{\sim}200$, W/V), color browning, reducing and total sugar, that were based on the significant levels of 10% of central composition design. Color browning, reducing and total sugar contents were found to be more affected when the water volume was increased rather than extraction time. Maximum extraction condition was acquired at extraction time of $3.0{\sim}4.5\;hrs$ and water volume of $40{\sim}58.2\;ml$. Being extracted at the optimal extraction condition two of the free sugars, sucrose (0.126%) and glucose (0.012%), were detected. Total content of the free amino acids was found to be $503.26\;{\mu}g%$. Among them, essential amino acid contents were revealed as 5.4%. One major peak from gel permeation chromatography contained polysaccharide(s) with the molecular weights of 10 KDa.

• Applied Biological Chemistry
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• v.28 no.3
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• pp.187-195
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• 1985

Electrophoretic studies of protein extracts from carrot calluses suspension-cultured on the media containing kinetin, BA, IAA, NAA or $GA_3$ at the levels of $10^{-6},\;10^{-5},\;10^{-4}M$, respectively, were performed to identify polypeptides and proteins regulated by auxin, cytokinin or GA. Fifteen bands of polypeptide(s) were observed in the callus cultured in the control medium devoid of growth regulators, and their molecular weights were $18._4,\;20._2,\;20._0,\;34._9,\;35._7,\;37._4,\;40._3,\;42._2,\;44._1,\;44._4,\;49._3,\;55._0,\;56._6,\;58._1,\;and\;59._9\;KD$, respectively. The synthesis of polypeptide appeared to be promoted in two bands by kinetin, in six bands by BA, in one band by IAA, in two bands by NAA, and in four bands by $GA_3$, while inhibited in five bands by kinetin, in three bands by BA, in four bands by IAA, in three bands by NAA and in three bands by $GA_3$. The polypeptides of $40._3\;KD\;42._2\;KD$ seemed to be regulated by cytokinins, and those of $44._1\;KD,37._4\;KD,\;and\;56._6\;KD$ by auxins. The proteins of three bands with relative mobilities of 0.56, 0.84, and 0.92, respectively, increased in the calluses cultured on the media containing kinetin, IAA, $GA_3$, NAA or BA, compared to the control, but it was difficult to identify the proteins specific for each growth regulator.

• The Korean Journal of Mycology
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• v.23 no.4 s.75
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• pp.332-339
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• 1995

Water-soluble polysaccharide (FCW) was extracted from the fruiting body of Fomitella fraxinea with neutral sodium chloride solution. The polysaccharide was further fractionated into FCW-I and FCW-II by ion exchange chromatography. The FCW-I and FCW-II were then purified by gel permeation chromatography and named as FCW-Ia and FCW-IIa, respectively. FCW-IIa showed relatively strong immuno-stimulating activity but FCW-Ia did not. By analyses of HPLC and GPC, FCW-Ia and FCW-IIa were identified to be homogeneous and their molecular weights were estimated to be about 15,000 and 8,700, respectively. FCW-Ia consisted of fucose, galactose, and mannose as main sugars and their molar ratio was 19.5 : 63.2 : 25.0. Protein was not detected in FCW-Ia. However, FCW-IIa was composed of glucose, galactose, and mannose at a molar ratio of 1.0 : 0.3 : 0.4 and contained 0.4% protein with a higher amount of glutamic acid. A small amount of uronic acid was detected in both FCW-Ia and FCW-IIa.

• Journal of Life Science
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• v.23 no.2
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• pp.273-281
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• 2013

The purpose of this research was to investigate the production and characterization of alkaline protease from Micrococcus sp. PS-1 newly isolated from seawater. Micrococcus sp. PS-1 was grown in Luria-Bertani (LB) medium. Its optimal temperature and pH for growth were $30^{\circ}C$ and 7.0, respectively. The effect of nitrogen sources was investigated on optimal enzyme production. A high level of alkaline protease production occurred in LB broth containing 2% skimmed milk. The protease was purified in a 3-step procedure involving ultrafiltration, acetone precipitation, and dialysis. The procedure yielded a 16.43-purification fold, with a yield of 54.25%. SDS-PAGE showed that the enzyme had molecular weights of 35.0 and 37.5 kDa. Its maximum protease activity was exhibited at pH 9.0 and $37^{\circ}C$, and its activity was stable at pH 8.0-11.0 and $25-37^{\circ}C$. The protease activity was strongly inhibited by PMSF, EDTA, and EGTA. Taken together, the results demonstrate that the protease enzyme from Micrococcus sp. PS-1 probably belongs to a subclass of alkaline metallo-serine proteases.

• The Korean Journal of Food And Nutrition
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• v.18 no.4
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• pp.309-315
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• 2005

The objective of this study was to investigate an influence of chitosan addition on quality attributes of sponge cake. The cake was made with various chitosan concentration (1,000, 2,000, and 3,000 ppm), and it was stored for 120 hr at three temperatures (5, 15, and $25^{\circ}C$). By analyzing the moxogram data of wheat flour-chitosan composite flour, the ascending angle of the samples containing chitosan was sharply increased and the descending angle had lower values compared to that of control as the chitosan addition concentration was increased. The loaf weights and volumes of the samples with chitosan were increased compared to those of the control. Crumb firmness of the control showed relatively high value compared to those ($5\~6$ N) of other samples containing chitosan regardless of storage temperature and storage period, and the firmness after 5 days storage at $5^{\circ}C$ showed the highest value (about 9.3 N). Sensory evaluation attributes of sponge cakes did not show significant differences by chitosan addition up to concentration of 2,000 ppm.

• Korean journal of food and cookery science
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• v.19 no.5
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• pp.616-623
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• 2003

Waxy barley brans were collected during the pearling process. The extraction of $\beta$-glucan from barley bran was effected by the extraction conditions. The $\beta$-glucan content increased with temperature, but not with pH. The highest yield, 6.5%, was achieved at pH 7.0 and 55$^{\circ}C$. At pH 10 and 45$^{\circ}C$, 48.5% of the $\beta$-glucan in barley bran was recovered in the gum product, with 54.6% purity. The protein and starch contaminations were high, reaching 13.6 and 23.7%, respectively. The $\beta$-glucan content was greatest in the subaleurone and aleurone regions (bran fractions 1, 2, 3 and 4), and declined considerably toward the inner layers. A monosaccharide analysis of the purified, $\beta$-glucan, from bran fractions 1, 2, 3 and 4, indicated that glucose constituted the majority of the gum. The small amounts of the arabinose and xylose found in the gum may indicate the presence of arabinoxylans as minor constituents. The molecular weights of the $\beta$-glucans isolated from bran fractions 1,2 and 3 were found to be 4.09${\times}$10$^{5}$ ∼-4.41${\times}$10$^{5}$ . The major glycosidic linkages of the $\beta$-glucans demonstrated the presence of 2, 4, 6-Me-Glc and 2, 3, 6-Me-Glc. When flow behaviors of barley bran $\beta$-glucan were examined, $\beta$-glucan exhibited pseudoplastic fluid properties.