• Title/Summary/Keyword: -cyclodextrin

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Isolation of Amylolytic Bifidobacterium sp. Int-57 and Characterization of Amylase

  • Ji, Geun-Eog;Han, Hee-Kyung;Yun, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.85-91
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    • 1992
  • The intestinal microflora of humans is an extraordinarily complex mixture of microorganisms, the majority of which are anaerobic microorganisms. The distribution of amylolytic microorganisms in the human large intestinal tract was investigated in various individuals of differing ages using anaerobic culture techniques. A large percentage of the amylolytic microorganisms present belonged to the Genus Bifidobacteria. The number of Bifidobacteria increased significantly at two years of age. Adults and children above 2 years old carried about $0.8{\times}10^9-2.0{\times}10^{10}$ colony forming units (CFU/gram) of amylolytic Bifidobacteria. Among these amylolytic Bifidobacteria, Int-57 was chosen for further studies. Between 65% and 85% of the amylase produced was secreted and the remaining amylase was bound to the cell wall facing the outside. Amylase production could be induced by starch in a stable form. When cells were grown on maltose or glucose, amylase production was much lower than on starch and amylase activity disappeared after 24 hours growth on these media. Partially purified enzymes showed optimum activity at a temperature of $50^{\circ}C$ and at an optimum pH of 5.5, respectively. Heat treatment at $70^{\circ}C$ for 30 minutes almost completely inactivated amylase. The hydrolysis products of starch were mainly maltose and maltotriose. Soluble starch, amylose, amylopectin, and $\gamma$-cyclodextrin($\gamma$-CD) were easily hydrolyzed. The rate of hydrolysis of $\alpha$-CD and $\beta$-CD was slower than that of $\gamma$-CD. Carboxymethyl cellulose, $\beta$-1, 3-glucan and inulin were not hydrolyzed.

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EFFECT OF CAPSAICIN AND EUGENOL ON ICGRP (IMMUNOREACTIVE CALCITONIN GENE-RELATED PEPTIDE) RELEASE FROM RAT LUMBAR SPINAL CORD. (백서 척수에서 Capsaicin과 Eugenol이 iCGRP (immunoreactive calcitonin gene-related peptide) 분비 조절에 미치는 영향.)

  • 오원만;김원재;최남기;박상원;황인남;김선헌
    • Restorative Dentistry and Endodontics
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    • v.26 no.5
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    • pp.436-442
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    • 2001
  • Neuropeptide such as calcitonin gene-related peptide and substance P may mediate neurogenic inflammation, but little is known about the regulation of neuropeptide release from rat spinal cord. Eugenol has been reported to reduce odontogenic pain and is known to have a structure similar to capsaicin, a potent stimulant of certain nociceptors. This study was done to examine the effect of capsaicin and eugenol on immunoreactive calcitonin gene-related peptide (iCGRP) release from rat spinal cord and whether eugenol regulates capsaicin-sensitive release of iCCRP or it evokes capsaicin-sensitive release of iCGRP. The dor-sal half of rat lumbar spinal cord was chopped into 200$\mu$m slices. They were superfused (500$\mu$l/min) in vitro with an oxygenated Kreb's buffer. The EC$_{50}$ of capsaicin on iCGRP release was measured. Eugenol (600$\mu$M and 1.2mM) and vehicle (0.02% 2-hydroxyl-$\beta$-cyclodextrin) were administered prior to stimulation of rat lumbar spinal cord with capsaicin. The amount of iCGRP release from rat lumbar spinal cord was measured by radioimmunoassay. The results were as follows : 1. iCGRP release from rat lumbar spinal cord was dependent on concentration of capsaicin. The EC$_{50}$ of capsaicin on iCGRP release was 3$\mu$M. 2. In the vehicle treated group, capsaicin (3$\mu$M) evoked a 14-fold increase over basal iCGRP level. 3. Administration of 600$\mu$M and 1.2mM eugenol evoked a 2.2-fold increase and a 2.3-fold increase over basal iCGRP level respectively. 4. Administration of 600$\mu$M and 1.2mM eugenol increased capsaicin evoked release of iCGRP by more than 50%. These results indicate that eugenol evoke CGRP release from central nervous system and potentiate the pain-inducing action of capsaicin on it.

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Studies on Dissolution of Fentiazac from ${\beta}-Cyclodextrin$ Inclusion Complex (${\beta}$-씨클로덱스트린 포접화합물로부터 펜티아작의 용출)

  • Yoon, Hyung-Joong;Back, Un-Bong;Seo, Seong-Hoon;Kim, Soo-Uck
    • Journal of Pharmaceutical Investigation
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    • v.20 no.3
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    • pp.153-159
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    • 1990
  • To increase the solubility of fentiazac which is used widely as a non-steroidal antiinflammatory drug, its inclusion complex and suppositories were prepared and studied. Inclusion complexes of fentiazac with ${\beta}-cyclodertin$ $({\beta}-CyD)$ were prepared by four diffrent methods; coprecipitation method, kneading method, solvent evaporation method, freeze drying method. Suppositories of $fentiazac/{\beta}-CyD$ with PEG 1500 and Witepsol H-15 were prepared by solvent evaporation method and freeze drying method. Inclusion complex formation of fentiazac with ${\beta}-CyD$ was ascertained by powder X-ray diffractometry, differential scanning calorimetry and IR spectroscopy. The dissolution rate of fentiazac from the inclusion complex increased in distilled water and KP 2nd disintegration test fluids (pH 6.8) but extemly decreased in KP 1st disintegration test fluid (pH 1.2). Inclusion complexes prepared by freeze drying method and solvent evaporation method were similar. Freeze drying method seemed to be suitable for preparation of complex with most higher dissolution rate but coprecipitation method seemed not to be suitable. The dissolution rate of fentiazac increased markedly by ${\beta}-CyD$ complexation. The release rates of suppositories increased in the following order. Complex prepared by freeze dying method in PEG 1500 > complex prepared by solvent evaporation method in PEG 1500 > fentiazac in PEG 1500 > complex prepared by freeze dying method in Witepsol H-15 > complex prepared by solvent evaporation method in Witepsol H-15 > fentiazac in Witepsol H-15.

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Expression of Cyclodextrinase Gene from Paenibacillus sp. A11 in Escherichia coli and Characterization of the Purified Cyclodextrinase

  • Kaulpiboon, Jarunee;Pongsawasdi, Piamsook
    • BMB Reports
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    • v.37 no.4
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    • pp.408-415
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    • 2004
  • The expression of the Paenibacillus sp. A11 cyclodextrinase (CDase) gene using the pUC 18 vector in Escherichia coli JM 109 resulted in the formation of an insoluble CDase protein in the cell debris in addition to a soluble CDase protein in the cytoplasm. Unlike the expression in Paenibacillus sp. A11, CDase was primarily observed in cytoplasm. However, by adding 0.5 M sorbitol as an osmolyte, the formation of insoluble CDase was prevented while a three-fold increase in cytoplasmic CDase activity was achieved after a 24 h-induction. The recombinant CDase protein was purified to approximately 14-fold with a 31% recovery to a specific activity of 141 units/mg protein by 40-60% ammonium sulfate precipitation, DEAE-Toyopearl 650 M, and Phenyl Sepharose CL-4B chromatography. It was homogeneous by non-denaturing and SDS-PAGE. The enzyme was a single polypeptide with a molecular weight of 80 kDa, as determined by gel filtration and SDS-PAGE. It showed the highest activity at pH 7.0 and $40^{\circ}C$. The catalytic efficiency ($k_{cat}/K_m$) values for $\alpha$-, $\beta$-, and $\gamma$-CD were $3.0{\times}10^5$, $8.8{\times}10^5$, and $5.5{\times}10^5\;M^{-1}\;min^{-1}$, respectively. The enzyme hydrolyzed CDs and linear maltooligosaccharides to yield maltose and glucose with less amounts of maltotriose and maltotetraose. The rates of hydrolysis for polysaccharides, soluble starch, and pullulan were very low. The cloned CDase was strongly inactivated by N-bromosuccinimide and diethylpyrocarbonate, but activated by dithiothreitol. A comparison of the biochemical properties of the CDases from Paenibacillus sp. A11 and E. coli transformant (pJK 555) indicates that they were almost identical.

Production of Glucosyl-xylitol Using Encapsulated Whole Cell CGTase (캡슐 고정화 전세포 CGTase를 이용한 Glucosyl-xylitol 생산)

  • 박중곤;박형우;이용현
    • KSBB Journal
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    • v.15 no.1
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    • pp.35-41
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    • 2000
  • We tried to prepare encapsulated whole cell cyclodextrin glucanotransferase(CGTase) in order to produce glycosyl-xylitol using xylitol as glucosyl acceptor. The organic nitrogen source was more effective for the production of CGTase from Bacillus macerans IFO 3490 than the inorganic one. Most of the CGTase which had been produced during cultivation was excreted to the growth medium. B. macerans cells inocculated in the capsule failed to grow to the high cell density. Adsorbents such as activated charcoal, Sephadex and Amberite resins could not adsorb efficiently the CGTase from the broth solution. We obtained successfully the encapsulated whole cell CGTase by immobilizing the concentrated broth solution in the calcium alginate capsules. The encapsulated whole cell CGTase carried out the transglycosylation reaction which converts xylitol into glucosyl-xylitol using dextrin as glucosyl donor.

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Effect of Browning Inhibitor Treatment on Sliced Citron Storage (Citrus junos Sieb.) (갈변방지제 처리가 슬라이스 유자의 저장성에 미치는 영향)

  • Lee, Bo-Bae;Lee, Jin-Won;Park, Jae-Ok;Cho, Youn-sup;Nam, Seung-Hee
    • The Korean Journal of Food And Nutrition
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    • v.34 no.4
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    • pp.390-397
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    • 2021
  • The purpose of this study was to investigate at how the quality of citron changed during storage as a result of the browning inhibitor treatment. In the browning inhibitor treatment, Vit.C, Vit.C+NaCl, Vit.C+NaCl+CD substances were used. As a result of investigating the browning degree, Vit.C+NaCl+CD showed the lowest value of 0.76 when stored for 12 weeks. The 𝚫E of the chromaticity value indicated that significant color change occurred when the value was high. As the Vit.C+NaCl+CD mixture showed the lowest value of 46.01 at 25℃, it was found that browning did not occur much compared to other treatments. The change in polyphenol oxidase (PPO) activity of citron increased as browning progressed. Among the browning inhibitor solutions, Vit.C+NaCl+CD solution showed the lowest value 118.8 u/g at 25℃ after 12 weeks. Based on these findings, it seems that CD mixing solution can be used as a citron browning inhibitor.

Enantioselective electrophoretic behavior of lipoic acid in single and dual cyclodextrin systems

  • Le, Thi-Anh-Tuyet;Nguyen, Bao-Tan;Phan, Thanh Dung;Kang, Jong-Seong;Kim, Kyeong Ho
    • Analytical Science and Technology
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    • v.34 no.4
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    • pp.143-152
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    • 2021
  • Capillary electrophoresis (CE) is an effective technique to study chiral recognition because it offers flexibility in adjusting vital factors. Currently, various available cyclodextrins (CDs) can be employed for the chiral separation of numerous analytes. Herein, we investigate the enantioseparation behavior of lipoic acid enantiomers in various types of single and dual CD systems through CE. Additionally, several impacted CE parameters were optimized through the systematic investigation based on the design of experiment (DoE) concept for a single system comprising a heptakis (2,3,6-tri-O-methyl)-β-CD and a dual system containing the combination of the single CD with a sulfated-β-CD. Consequently, absolute enantioresolution was obtained within 15 min on a common standard bare fused-silica capillary (64.5/56 cm in total/effective length, 50/365 ㎛ inner/outer diameter), maintained at 15 ℃ and at an applied voltage of 24 kV. The optimal background electrolyte consisted of 6 mM heptakis (2,3,6-tri-O-methyl)-β-CD dissolved in the solution of 58 mM borate buffer at pH 10. Furthermore, the results of apparent binding constant experiments indicated that the S-enantiomer-heptakis (2,3,6-tri-O-methyl)-β-CD complex exhibited a stronger affinity than its R-enantiomer counterpart. The obtained electrophoretic mobility values could be utilized to interpret the resolution achieved at various CD concentrations and the mobility behavior of the complexes elucidated the migration order of the enantiomers in an electropherogram.

Reduction of 2,4,6-Trinitrotoluene Mobility in Operational Range Soil by Sorption Enhancement and Desorption Decrease Using Monopotassium Phosphate and Montmorillonite (제일인산칼륨과 몬트모릴로나이트 점토를 이용한 사격장 토양 내 2,4,6-trinitrotoluene의 흡착증진 및 탈착감소에 의한 이동성 저감 연구)

  • Jung, Jae-Woong;Yu, Gihyeon;Nam, Kyoungphile
    • Journal of Soil and Groundwater Environment
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    • v.23 no.6
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    • pp.46-53
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    • 2018
  • Mobility reduction of 2,4,6-trinitrotoluene (TNT) was tested by amending monopotassium phosphate (MKP) and montmorillonite to a firing range soil contaminated with TNT. While addition of MKP enhanced sorption of TNT on soil matrix, and combined use of MKP with montmorillonite significantly decreased desorption of TNT as well as remarkably increased the TNT sorption. Montmorillonite amendment by 5% of soil mass resulted in TNT desorption of 0.12 mg/kg from soil loaded with 9.93 mg/kg-TNT. The decrease of TNT desorption was proportional to the amount of montmorillonite amended. At 10 and 15% amendment, only 0.79 and 1.23 mg/kg-TNT was desorbed from 29.33 and 48.80 mg/kg-TNT. In addition, the leaching of TNT with synthetic precipitation leaching procedure (SPLP) and hydroxypropyl-${\beta}$-cyclodextrin (HPCD) decreased, indicating that TNT in MKP/montmorillonite-treated soil became more stable and less leachable. The results demonstrate that addition of MKP and montmorillonite to TNT-contaminated soil reduces the mobility of TNT from soil not only by increasing TNT sorption, but also decreasing TNT desorption. It was found that MKP and montmorillonite amendments by 5 and 10% of soil mass, respectively, were optimal for reducing the mobility of soil TNT.

Characterization of the Transglycosylation Reaction of 4-α-Glucanotransferase (MalQ) and Its Role in Glycogen Breakdown in Escherichia coli

  • Nguyen, Dang Hai Dang;Park, Sung-Hoon;Tran, Phuong Lan;Kim, Jung-Wan;Le, Quang Tri;Boos, Winfried;Park, Jong-Tae
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.357-366
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    • 2019
  • We first confirmed the involvement of MalQ (4-${\alpha}$-glucanotransferase) in Escherichia coli glycogen breakdown by both in vitro and in vivo assays. In vivo tests of the knock-out mutant, ${\Delta}malQ$, showed that glycogen slowly decreased after the stationary phase compared to the wild-type strain, indicating the involvement of MalQ in glycogen degradation. In vitro assays incubated glycogen-mimic substrate, branched cyclodextrin (maltotetraosyl-${\beta}$-CD: G4-${\beta}$-CD) and glycogen phosphorylase (GlgP)-limit dextrin with a set of variable combinations of E. coli enzymes, including GlgX (debranching enzyme), MalP (maltodextrin phosphorylase), GlgP and MalQ. In the absence of GlgP, the reaction of MalP, GlgX and MalQ on substrates produced glucose-1-P (glc-1-P) 3-fold faster than without MalQ. The results revealed that MalQ led to disproportionate G4 released from GlgP-limit dextrin to another acceptor, G4, which is phosphorylated by MalP. In contrast, in the absence of MalP, the reaction of GlgX, GlgP and MalQ resulted in a 1.6-fold increased production of glc-1-P than without MalQ. The result indicated that the G4-branch chains of GlgP-limit dextrin are released by GlgX hydrolysis, and then MalQ transfers the resultant G4 either to another branch chain or another G4 that can immediately be phosphorylated into glc-1-P by GlgP. Thus, we propose a model of two possible MalQ-involved pathways in glycogen degradation. The operon structure of MalP-defecting enterobacteria strongly supports the involvement of MalQ and GlgP as alternative pathways in glycogen degradation.

Flame-retarding effects depending on the number of phosphonate groups attached to phosphorus flame-retarding compounds and coating binder resins (인계 난연화합물 및 코팅 바인더 수지에 부착된 phosphonate group에 따른 난연효과)

  • Park, Hyo-Nam;Kim, Hae-Rim;Choi, Seong-Ho
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.6
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    • pp.1678-1686
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    • 2021
  • In this study, we prepared phosphorous flame-retarding coating solutions by mixing triphosphate (3 phosphonate), phytic acid (6 phosphonate), or ammonium polyphosphate (10 phosphonate) with boric acid as a crosslinking agent and acryl resin binder. Prepared phosphorous flame-retarding coating solutions were coated onto non-woven fabrics, respectively, to obtain high flame-retarding effects. These prepared flame-retardant non-woven fabrics were evaluated using smoke density standard test (ASTM E662), limit oxygen index standard test (ISO E622), and vertical burning standard test (UL 94). Their flame-retarding effects were affected by the number of phosphonate groups. Regardless of natural or synthetic binder resins, their effects showed the following order: ammonium polyphosphate > phytic acid > triphosphate. Natural hydrocarbon compounds were also examined to determine the possible retardancy of binder resins. Results showed that natural hydrocarbon binder resins could be used for preparing fire-retardant nonwoven fabrics.