• Title/Summary/Keyword: low molecular fraction

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Purification and Biochemical Characterization of Sucrose Synthase from the Cytosolic Fraction of Chickpea (Cicer arietinum L. cv. Amethyst) Nodules

  • Lee, Hoi-Seon
    • Journal of Applied Biological Chemistry
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    • v.42 no.1
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    • pp.12-18
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    • 1999
  • Sucrose synthase (EC 2.4.1.13) has been purified from the plant cytosolic fraction of chickpea (Cicer arietinum L. cv. Amethyst) nodules. The native enzyme had a molecular mass of $356{\pm}15kD$. The subunit molecular mass was $87{\pm}2kD$, and a tetrameric structure is proposed for sucrose synthase of chickpea nodule. Optimum activities in the sucrose cleavage and synthesis directions were at pH 6.5 and 9.0, respectively. The purified enzyme displayed typical hyperbolic kinetics with substrates in cleavage and synthesis reactions. Chickpea nodules sucrose synthase had a high affinity for UDP ($K_m$, $8.0{\mu}M$) and relatively low affinities for ADP ($K_m$, 0.23 mM), CDP ($K_m$, 0.87 mM), and GDP ($K_m$, 1.51 mM). The $K_m$ for sucrose was 29.4 mM. In the synthesis reaction, UDP-glucose ($K_m$, $24.1{\mu}M$) was a more effective glucosyl donor than ADP-glucose ($K_m$, 2.7 mM), and the $K_m$ for fructose was 5.4 mM. Divalent cations, such as $Ca^{2+}$, $Mg^{2+}$, and $Mn^{2+}$, stimulated the enzyme activity in both the cleavage and synthesis directions, and the enzyme was very sensitive to inhibition by $HgCl_2$ and $CuSO_4$.

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A Simple and Rapid Method to Isolate Low Molecular Weight Proteinase Inhibitors from Soybean

  • Krishnan Bari B.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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    • pp.342-348
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    • 2004
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the $60\%$ isopropanol extract of soybean(Glycine max [L.] Merr.) seed revealed two abundant proteins with molecular masses of 19 and 10 kDa. Amino acid analysis revealed that the isopropanol-extractable protein fraction was rich in cysteine. Two-dimensional gel electro-phoretic analysis indicated that the 19kDa and 10kDa proteins had pI of 4.2 and 4.0 respectively. Peptide mass fingerprints of trypsin digests of the two proteins obtained using matrix-assisted, laser desorption/ionization-time of flight (MALDI-TOF) mass spectroscopy revealed the 19kDa protein was Kunitz trypsin inhibitor and the 10kDa protein was Bowman-Birk proteinase inhibitor. When resolved under non-denaturing conditions, the isopropanol-extracted proteins inhibited trypsin and chymotrypsin activity. Results presented in this study demonstrate that isopropanol extraction of soybean seed could be used as a simple and rapid method to obtain a protein fraction enriched in Kunitz trypsin and Bowman-Birk proteinase inhibitors. Since proteinase inhibitors are rich in sulfur amino acids and are putative anticarcinogens, this rapid and inexpensive isolation procedure could facilitate efforts in nutrition and cancer research.

The Effect of Polyphenol Oxidase on Deodorizing Activity of Apple Extract against Methyl Mercaptan (사과 추출물의 구취억제효과에 대한 Polyphenol Oxidase의 영향)

  • 조상원;곽기석;이주항;윤영수;구연숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1301-1304
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    • 2001
  • Deodorizing activity of polyphenol cxidase (PPO) extracted from apples was investigated by measuring the changes of methyl mercaptan as an indicator of halitosis in human mouths. In the studies of apple extracts on deodorizing activity, the deodorizing activity was increased with the amount of apple extracts. In the cases of adding PPO to the low molecular fraction of apple extracts, the deodorizing activities were increased with the amount of the law molecular fraction of apple extracts and the reaction time of the extracts with PPO. Deodorizing activities of PPO is thought that o-quinone as an intermidiate produced by an oxidative reaction of PPO during enzymatic browning reactions may react with methyl mercaptan to form a non-volatile and sulfur-containing compound .

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Anti-aging potential of fish collagen hydrolysates subjected to simulated gastrointestinal digestion and Caco-2 cell permeation

  • Je, Hyun Jeong;Han, Yoo Kyung;Lee, Hyeon Gyu;Bae, In Young
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.101-107
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    • 2019
  • The objectives of this study were to evaluate the anti-aging effects and investigate the effect of simulated gastrointestinal (GI) digestion on the anti-aging properties and intestinal permeation of the potential fish collagen hydrolysates (FCH). Therefore, procollagen synthesis, matrix metalloproteinase-1 (MMP-1) production, and Caco-2 cell permeability were analyzed before and after in vitro digestion for FCHs, low-molecular weight fractions (<1 kDa), and high molecular weight fractions (>1 kDa). After being subjected to GI digestion, the level of MMP-1 inhibition was maintained, although the procollagen production was significantly (>20%) lower with all samples. Also, the digested FCHs and their <1 kDa fraction yielded 9.1 and 13.8% increased peptide transport, respectively, compared to undigested samples. Based on the effective intestinal permeation and high digestive enzyme stability, the <1 kDa fraction of FCHs is a potential bioactive material suitable for anti-aging applications in the food and cosmetics industries.

Study on the Molecular Weight Distribution Curve of Cellulose Triacetate Acetylated Under Various Temperatures (醋酸纖維素의 醋化溫度가 分子量分配曲線에 미치는 影響)

  • Kim, Dong-Il;Noh, Ick-Sam;Cha, Kyong-Mo
    • Journal of the Korean Chemical Society
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    • v.4 no.1
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    • pp.44-50
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    • 1957
  • Fibrous cellulose triacetate prepared from purified cotton under various temperatures was dissolved in the solution of 70%, monochloroacetic acid and it was fractionated using water as a precipitant. Eight fractions were obtained through the stepwise precipitation. Degree of polymerization and molecular weight of each fraction were measured viscometrically. Integral and differential molecular weight distribution curve were drawn for each sample prepared under various temperatures and were carefully observed. On this experimental study, following conclusions were obtained: Fractional precipitation can be carried out for fibrous cellulose triacetate in the solution of 70% monochloroacetic acid using water as a precipitant. The differences on the shapes of molecular weight distribution curve were occured on account of the various acetylation temperatures. At the relatively higher acetylation temperatures, the cellulose was randomly degraded and the portion of low degree of polymerization was increased. Commercial acetate, therefore, may not be prepared at above 40$^{\circ}C$ according to the molecular weight distribution curve regardless of higher viscosity and average degree of polymerization. It was concluded that the optimum acetylation temperature for commercial acetate was approximately 30$^{\circ}C$.

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Softening Related Changes in Cell Wall Polysaccharides of Persimmon (감의 연화와 관련된 세포벽다당류의 변화)

  • Kim, Soon-Dong;Park, Nam-Sook;Kang, Meung-Su
    • Korean Journal of Food Science and Technology
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    • v.18 no.2
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    • pp.158-162
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    • 1986
  • Various cell wall polysaccharides such as ionically associated pectin (IAP), covalently bounded pectin (CBP),4N potassium hydroxide soluble hemicellulosic fraction (HF,) and 0-3N soluble hemicellulosic fraction (HF,) were fractionated from crude cell wall of the fresh and soft persimmon by chemical method. The changesin cell wall polysaccharides were studied by gel filteration chromatography . The content of crude cell wall remarkably decreased in the soft persimmon. The decreasing rates of IAP, CBP and $HF_2$ were 59, 60 and 74%, respectively, while $HF_1$ and cellulose changed only a little during softening. Sugar compositions of IAP and CBP were 72-84% uronic acid, 5-1% hexose and 11-16% pentose, and also the hemicellulose was composed of uronic acid besides hexose and pentose that was hemicellulosic components. The loss rate of pentose in IAP, of hexose in CBP, of hexose and uronic acid in $HF_2$, of pentose in $HF_1$ increased during softening. Though apparent average molecular freight of all polysaccharides shifted from high molecular freight to low molecular weight polymer, the shifting degree of CBP and $HF_2$ was especially remarkable during softening. It is suggested that the severe softening phenomenon of persimmon involved the degradation and dissolution of wall bound-CBP and $HF_2$ which were associated with each other.

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Isolation and Characterization of Complement System Activating Polysaccharides from the Hot Water Extract of the Leaves of Capsicum annuum L. (고추잎 열수추출물로부터 보체계 활성화 다당의 분리 및 특성)

  • 이용세;나경수;정상철;서형주;박희성;백형석;최장원
    • Journal of Life Science
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    • v.12 no.1
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    • pp.87-95
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    • 2002
  • It was observed that the hot water extract of the leaves of Capsicum annuum L., a Korean edible plant, had a potent anti-complementary activity. Crude polysaccharide fraction(CAL-0) was obtained by methanol reflux, ethanol precipitation, dialysis and lyophilization. CAL-0 contained 51.8% of total sugar, 8.2% of uronic acid and 16.8% of protein, and consisted of mainly arabinose, galactose and glucose as neutral sugars and galacturonic acid as uronic acid. The anti-complementary activity of CAL-0 decreased greatly by periodate oxidation, but was not changed by pronase treatment. Also, the anti-complementary activity of CAL-0 was reduced partially in the absence of the $Ca^{2+}$ ion. The crude polysaccharide CAL-0 was found to activate the C3 component both in the presence and in the absence of $Ca^{2+}$ through the crossed-immunoelectrophoresis suggesting that those involved in both classical and alternative complement pathway CAL-0 was further separated to an unabsorbed fraction(CAL-1) and six absorbed fractions(CAL-2longrightarrowCAL-7) on DEAE Sepharose CL-6B ion exchange column. Among them four major fractions in activity and yield were obtained, and consisted mainly of arabinose, galactose and glucose with various molar ratios. The major fraction, CAL-2, was purified to give a high molecular fraction(CAL-2-I) and a low molecular fraction(CAL-2-II) on Sepharose CL-6B column. The anti-complementary activity of CAL-2-I, a molecular weight of about 61,000, was higher than it of CAL-2-II.-II.

Effects of Egg Shell Membrane Hydrolysates on Anti-Inflammatory, Anti-Wrinkle, Anti-Microbial Activity and Moisture-Protection

  • Yoo, Jinhee;Park, Kimoon;Yoo, Youngji;Kim, Jongkeun;Yang, Heejin;Shin, Youngjae
    • Food Science of Animal Resources
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    • v.34 no.1
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    • pp.26-32
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    • 2014
  • This study was conducted to examine the effects of eggshell membrane hydrolysates (ESMH) on the anti-inflammatory, anti-wrinkle, anti-microbial activity, and moisture-protection for cosmetic use. Whole ESMH (before fractionation), and fraction I (>10 kDa), fraction II (3-10 kDa), and fraction III (<3 kDa) of the hydrolysates were assessed in this experiment. As lipopolysaccharide (LPS) and IFN-${\gamma}$ caused the inflammation on Raw264.7 cell, whole ESMH and fraction I showed to be effective in inhibiting the induction of cell inflammation depending on the concentration, and also showed outstanding effect to suppress the skin inflammation. Fraction I inhibited collagenase and elastase activities to a greater extent than the other fractions, while all fractions had antibiotic effects at concentrations of 10 mg/disc and 20 mg/disc. In addition, it showed the moisture protection effects of skin on the holding amount and losing amount of moisture in upper-inner arm of the human body with a relatively low loss rate in skin, which confirmed that the hydrolyzed fractions of ESM helps to form the superior protective layer of moisture. It was concluded that ESMH fractions with different molecular weights, especially the 10 kDa fraction, have anti-lipopolysaccharide, anti-IFN-${\gamma}$-induced inflammation, anti-collagenase and elastase activities, and thus can be used as a cosmetic agent to protect skin.

In vivo Angiogenic Activity of Dichloromethane Extracts of Aloe vera Gel

  • Lee, Myoung-Jin;Yoon, Soo-Hong;Lee, Seung-Ki;Chung, Myung-Hee;Park, Young-In;Sung, Chung-Ki;Choi, Jae-Sue;Kim, Kyu-Won
    • Archives of Pharmacal Research
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    • v.18 no.5
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    • pp.332-335
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    • 1995
  • The angiogenic activity of Aloe vera (Aloe baradensis), known as a good healing plant, was investigated. We have extracted and fractionated dichloromethane extract (G1M1D1) and methanol soluble fraction of dichloromethane extract (G1M1D1M1) which contain low-molecular weight substances of Aloe vera gel. G1M1D1 and G1D1M1 fractions induced a radially arranged, spoke-sheel-like vasculature in chick embryo chorioallantoic membrane (CAM) assay. The angiogenic activity was dose-dependent and the angiogenic pattern in the CAM assay. The angiogenic activity was dose-dependent and the angiogenic pattern in the CAM assay was very similar to that of phorbol 12-myristate-13-acetate (PMA) used as a positive control. The modified CAM assay, e simple and accurate quantitating method, was used to quntitate the angiogenic activity of G1D1M1 fraction. Application of G1M1D1M1 fraction ($100\mug/egg$) resulted in much more intense angiogenesis than in control while slightly less intense angiogenesis than in PMA (100 ng/egg).

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Characteristics and Dyeing Properties of Green Tea Colorants(Part I) -Components and characteristics of Green Tea Colorants- (녹차색소의 특성과 염색성 (제1보) -녹차색소의 성분과 특성-)

  • Shin, Youn-sook;Choi, Hee
    • Journal of the Korean Society of Clothing and Textiles
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    • v.23 no.1
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    • pp.140-146
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    • 1999
  • Colorants in green tea were extracted freeze-dried and analyzed to investigate the possibility of using as a natural dye. Fractionation of the colorants was carried out by column chromatograpy. Colorants in green tea were eluted into five fractions. All the fractions except fraction F2 showed absorption peakat 280nm. Fraction f2 showed absorption peak at 270nm and broad peak at 350nm, From the IR analysis it is speculated that fractions F2-F5 having similar stucture but different molecular weight are catechis. Silk fabrics dyes with fractions F1-F4 showed yellowish red color while sample dyed with fraction F5 showed red color. The colorants from green tea infusion was applied to silk wool nylon cotton and rayon fabrices. It showed relatively good affinity to protein and polyamide fibers bur low affinity to cellulose and regenerated cellulose fibers.

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