• Title/Summary/Keyword: hydrolysis patterns

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Effect of High Pressure on the Porcine Placenral Hydrolyzing Activity of Pepsin, Trypsin and Chymotrypsin

  • Chun, Ji-Yeon;Jo, Yeon-Ji;Min, Sang-Gi;Hong, Geun-Pyo
    • Food Science of Animal Resources
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    • v.34 no.1
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    • pp.14-19
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    • 2014
  • This study investigated the effects of protease treatments (trypsin, chymotrypsin, and pepsin) under various pressure levels (0.1-300 MPa) for the characteristics of porcine placenta hydrolysates. According to gel electrophoretic patterns, the trypsin showed the best placental hydrolyzing activity followed by chymotrypsin, regardless of the pressure levels. In particular, the peptide bands of tryptic-digested hydrolysate were not shown regardless of applied pressure levels. The peptide bands of hydrolysate treated chymotrypsin showed gradual decreases in molecular weights ($M_w$) with increasing pressure levels. However, the pepsin did not show any evidences of placental hydrolysis even though the pressure levels were increased to 300 MPa. The gel permeation chromatography (GPC) profiles showed that the trypsin and pepsin had better placental hydrolyzing activities under high pressure (particularly at 200 MPa), with lower $M_w$ distributions of the hydrolysates. Pepsin also tend to lower the $M_w$ of peptides, while the major bands of hydrolysates being treated at 300 MPa were observed at more than 7,000 Da. There were some differences in amino acid compositions of the hydrolysates, nevertheless, the peptides were mainly composed of glycine (Gly), alanine (Ala), hydroxyproline (Hyp) and proline (Pro). Consequently, the results indicate that high pressure could enhance the placental hydrolyzing activities of the selected proteases and the optimum pressure levels at which the maximum protease activity is around 200 MPa.

Molecular Structural Properties of Waxy Rice Starch (찹쌀 전분의 분자 구조적 성질)

  • Kim, Kwan;Choi, Gyeong-Chul;Kang, Kil-Jin;Lee, Yong-Hyun;Kim, Sung-Kon
    • Korean Journal of Food Science and Technology
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    • v.24 no.6
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    • pp.568-573
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    • 1992
  • Molecular structural properties of Japonica and Tongil type waxy rice starches were compared. The general properties including water-binding capacity, swelling power at $80^{\circ}C$, intrinsic viscosity, leach loss at $98^{\circ}C$ and ${\beta}-amylolysis$ limit were similar but the chain length was slightly different between the two starches. Similar elution profiles on Sephadex G-50 were obtained when the starches were treated with ${\beta}-amylase$. The pullulanase treated starches however showed different elution patterns each other. The ${\beta}-amylolysis$ of acid-treated (2 days) starches linearly increased as hydrolysis time prolonged. When the acid-treated starches were hydrolyzed with ${\beta}-amylase$ or pullulanase, the elution profiles on Sepharose CL-2B were considerably different from those of native starches.

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Stability of Soybean Isoflavone Isomers According to Extraction Conditions

  • Choi, Yeon-Bae;Kim, Kang-Sung
    • Journal of Environmental Health Sciences
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    • v.31 no.6
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    • pp.498-503
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    • 2005
  • Stability of soybean isoflavone isomers according to extraction conditions such as temperature, pH, and extracting solvents was investigated. Heating induced three chemical reactions to occur for malony1 derivatives of isoflavones, namely decarboxylation of malony1 groups into acety1 derivatives, deesterification of malony1 residues, and hydrolysis of $\beta$-glycosidic bonds. Among the twelve isoflavone isomers, change in concentrations of acety1glycosides were most pronounced: Acety1 derivatives were present only in trace amounts in unheated hypocotyls, but the content increased dramatically during heating. As for the glycosides, concentrations of daidzin and glycitin increased due to heat treatment, though that of genistin remained almost unchanged. Heat decomposition rates and the patterns differed among the three malony1 derivatives. After 120 min at $80^{circ}C$, the relative concentrations of daidzin, glycitin and genistin were increased from $9.2\%$, $12.4\%$ and $3.3\%$ to $19.3\%$, $21.9\%$ and $6.2\%$, respectively. When crude isoflavones were solubilized in glycine buffer (pH 10.0) and incubated at $80^{circ}C$, deesterification occurred faster than at pH 7.0. When the pH of isoflavone solution was increased, the malony1glycosides were hydrolyzed to their respective glycosides at increased rate. Both acetyl and aglycone forms were unchanged and only de-esterification reactions occurred. At the acidic pH, malonylglycosides were much stable both at 60 and $80^{circ}C$. However at pH 10, $80^{circ}C$ and 1 hr, $75-80\%$ of malonylglycosides were transformed to their deesterified glycosides. When isoflavones were extracted with $60\%$ aqueous ethanol at $60^{circ}C$, isoflavone isomers were stable and the deesterification reactions did not occur in these conditions. However, at $80^{circ}C$ deesterification of malonyiglycosides occurred significantly with $15-20\%$ of malonylglycosides being hydrolyzed into their respective glycosides. This experiment showed that malonylglycosides undergo decomposition when heated or exposed to alkaline conditions. Also, aqueous ethanol was preferred to aqueous methanol as solubilizing media for obtaining extract with minimum degradation of malonylglycosides.

Separation of Calcium-binding Protein Derived from Enzymatic Hydrolysates of Cheese Whey Protein

  • Kim, S.B.;Shin, H.S.;Lim, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.712-718
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    • 2004
  • This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

Characterization of Campylobacters Newly Isolated from Swine Gastric Mucosa

  • Lee, Yeon-Hee;Lee, Jin-Hui;Cho, Hyun-Joo;Shin, En-Joo;Park, Jong-Hwan;Park, Jae-Hak
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.778-783
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    • 1999
  • Campylobacter is a pathogen for both humans and animals that can be transferred from animals to humans. Four isolates, which grew under 5-10% $CO_2$ and had small and translucent colonies, were obtained from swine gastric mucosa and characterized using various methods. These bacteria were gram negative, spirally shaped with round ends. One or two non-sheathed polar flagella were observed under electron microscopy. A PCR with species-specific protein (SSP) primers for 16S rRNA gene in Campylobacter produced a typical 462 bp fragment. The isolates had various biochemical and molecular characteristics which differentiated them from other Campylobacters. The isolates were catalase and oxidase positive, urease (rapid) negative, nitrate reduction positive, indoxyl acetate hydrolysis positive, y-glutamyl transpeptidase negative, and alkaline phosphatase negative. All four isolates showed growth at $37^{\circ}C{\;}and{\;}42^{\circ}C{\;}but{\;}not{\;}at{\;}25^{\circ}C$, were resistant to cephalotin and cefoperazone, and susceptible to carbenicillin. The isolates showed various results in the reduction of chloride to triphenyl tetrazolium (TTC) and a susceptibility to nalidixic acid. Western blot analysis of these isolates with antiserum raised against one isolate showed different patterns from those of reference strains. A dendrogram drawn with the RAPD results showed that these isolates belonged to a new Campylobacter spp. group different from those of C. jejuni, C. doylei, C. lari, and C. coli.

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Response Characteristics of Thick Film Sensors Using Nano ZnO:Ni for Hydrocarbon Gas (나노 ZnO:Ni를 이용한 후막 가스센서의 탄화수소계 가스에 대한 감응특성)

  • Yoon, So-Jin;Yu, Il
    • Korean Journal of Materials Research
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    • v.23 no.4
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    • pp.211-214
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    • 2013
  • The effects of a Ni coating on the sensing properties of nano ZnO:Ni based gas sensors were studied for $CH_4$ and $CH_3CH_2CH_3$ gases. Nano ZnO sensing materials were prepared by the hydrothermal reaction method. The Ni coatings on the nano ZnO surface were deposited by the hydrolysis of zinc chloride with $NH_4OH$. The weight % of Ni coating on the ZnO surface ranged from 0 to 10 %. The nano ZnO:Ni gas sensors were fabricated by a screen printing method on alumina substrates. The structural and morphological properties of the nano ZnO : Ni sensing materials were investigated by XRD, EDS, and SEM. The XRD patterns showed that nano ZnO : Ni powders with a wurtzite structure were grown with (1 0 0), (0 0 2), and (1 0 1) dominant peaks. The particle size of nano ZnO powders was about 250 nm. The sensitivity of nano ZnO:Ni based sensors for 5 ppm $CH_4$ gas and $CH_3CH_2CH_3$ gas was measured at room temperature by comparing the resistance in air with that in target gases. The highest sensitivity of the ZnO:Ni sensor to $CH_4$ gas and $CH_3CH_2CH_3$ gas was observed at Ni 4 wt%. The response and recovery times of 4 wt% Ni coated ZnO:Ni gas sensors were 14 s and 15 s, respectively.

Extracellular Secretion of a Maltogenic Amylase from Lactobacillus gasseri ATCC33323 in Lactococcus lactis MG1363 and its Application on the Production of Branched Maltooligosaccharides

  • Cho, Mee-Hyun;Park, Sang-Eun;Lee, Myung-Hun;Ha, Suk-Jin;Kim, Hae-Yeong;Kim, Myo-Jeong;Lee, Sung-Joon;Madsen, Soren M.;Park, Cheon-Seok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1521-1526
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    • 2007
  • A maltogenic amylase gene from Lactobacillus gasseri ATCC33323 (LGMA) was expressed in Lactococcus lactis MG1363 using the P170 expression system. The successful production of recombinant LGMA (rLGMA) was confirmed by the catalytic activity of the enzyme in liquid and solid media. The N-terminal amino acid sequencing analysis of the rLGMA showed that it was Met-Gln-Leu-Ala-Ala-Leu-, which was the same as that of genuine protein, meaning the signal peptide was efficiently cleaved during secretion to the extracellular milieu. The optimal reaction temperature and pH of rLGMA ($55^{\circ}C$ and pH 5, respectively) and enzymatic hydrolysis patterns on various substrates (${\beta}$-cyclodextrin, starch, and pullulan) supported that rLGMA was not only efficiently secreted from the Lactococcus lactis MG1363 but was also functionally active. Finally, the branched maltooligosaccharides were effectively produced from liquefied com starch, by using rLGMA secreted from Lactococcus lactis, with a yield of 53.1%.

The Fine Structure of Amylopectin and Physicochemical Properties of Starch Granules from Endosperm Varieties in Glutinous Rice (찹쌀 전분의 미세구조 및 이화학적 특성 비교)

  • Sung, You-Me;Nam, Seok-Hyun;Kang, Mi-Young
    • Applied Biological Chemistry
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    • v.43 no.1
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    • pp.18-23
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    • 2000
  • stract : Fourteen varieties of glutinous rices were examined on amylopectin fine structure and physicochemical properties of starch granules. The amylopectin chain length distribution and short chain/long chain ratio were investigated by enzymatic treatments followed by high-performance size-exclusion chromatographic separation. Chain length distribution profiles of the isoamylase-debranched amylopectins showed distinct patterns according to varieties. Beongok showed the highest short chain/long chain ratio, while TP2579A1 showed the lowest one. Sharebyeo-152-1-B showed the highest hydrolysis rate to 15% $H_2SO_4$, while Sandong 47 showed the lowest one. Fourteen varieties of rice starch granules showed A-type pattern on X-ray diffractograms. Non-gelitinized starch granules from Keochang 1 and Beongok had almost 100% hydrolysed by glucoamylase for 3 hrs at $370^{\circ}C$.

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Gene Expression Analysis of the Bromobenzene Treated Liver with Non-hepatotoxic Doses in Mice

  • Lim, Jung-Sun;Jeong, Sun-Young;Hwang, Ji-Yoon;Park, Han-Jin;Cho, Jae-Woo;Song, Chang-Woo;Kim, Yang-Seok;Lee, Wan-Seon;Moon, Jin-Hee;Han, Sang-Seop;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.268-274
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    • 2005
  • Bromobenzene (BB) is well known hepatotoxicant. Also, BB is an industrial solvent that arouses toxicity predominantly in the liver where it causes centrilobular necrosis. BB is subjected to Cytochrome P450 mediated epoxidation followed by either conjugation with glutathione, enzymatic hydrolysis or further oxidation. In this study, we focused on BB-induced gene expression at non-hepatotoxic dose. Mice were exposed to two levels of BB, sampled at 24 h, and hepatic gene expression levels were determined to evaluate dose dependent changes. When examining the toxic dose of BB treated group in other previous studies, genes related to heat shock protein, oxidative stress, and drug metabolism are expressed. Compared to these results, our study, in which non-toxic dose of BB was administrated, showed similar patterns as the toxic conditions above. The purpose of the study was to select genes that showed changes in relation to the differing dose through confirmation of the difference within transcriptomic boundaries, but those that are not detected by the existing classic toxicology tools in non-hepatotoxic dose.

Purification and Characterization of Xylanase I from Trichoderma koningii ATCC 26113 (Trichoderma koningii ATCC 26113으로부터 Xylanase 1의 순수분리 및 특성)

  • Kim, Hyun-Ju;Kang, Sa-Ouk;Hah, Yung-Chil
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.63-71
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    • 1993
  • A xylanase (xylanase I) was purified 11.9-fold from the culture filtrate of Trichoderma koningii ATCC 26113 by the column chromatography on Sephadex G-75, SP-Sephadex C-50, DEAE-Sephadex A-50 and Sephadex G-50 with an overall yield of 8.2%. The molecular mass determined by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis was found to be a monomeric polypeptide of ca. 35 kDa. The isoelectric point of the enzyme was estimated to be 9.3. The optimal reaction pH and temperature are 5.8 and 55.deg.C, respectively. The enzyme is stable up to 60.deg.C, while 78% of its activity is lost after the incubation for 10 min at 70.deg.C. The enzyme hydrolyzes sylan with relatively high activity, as well as carboxymethyl cellulose and avicel. The $K_{m}$ values of the enzyme for oat-spelf sylan, larchwood xylan and Avicel were 3.5, 1.6 and 10. 1 mg/ml, respectively. The enzyme hydrolyzed oat-spelt sylan to sylose, sylobiose, sylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose, xylotriose and arabionoxylobiose, while it degraded larchwood xylan to xylose, xylobiose and xylotriose as the major products. The hydrolysis patterns indicate that xylanase I is endo-enzyme.e.

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