• Title/Summary/Keyword: ${\alpha}-Cyclodextrin$

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Molecular Cloning and Characterization of Maltogenic Amylase from Deinococcus geothermalis (Deinococcus geothermalis 유래 maltogenic amylase의 유전자 발현 및 특성확인)

  • Jung, Jin-Woo;Jung, Jong-Hyun;Seo, Dong-Ho;Kim, Byung-Yong;Park, Cheon-Seok
    • Korean Journal of Food Science and Technology
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    • v.43 no.3
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    • pp.369-374
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    • 2011
  • A putative maltogenic amylase gene (DGMA) was cloned from the Deinococcus geothermalis DSM 11300 genome using the polymerase chain reaction. The gene encoded 608 amino acids with a predicted molecular mass of 68,704 Da. The recombinant DGMA was constitutively expressed using the pHCXHD plasmid. As expected, the recombinant DGMA hydrolyzed cyclodextrins and starch to maltose and pullulan to panose by cleaving the ${\alpha}$-(1,4)-glycosidic linkages, as observed for typical maltogenic amylases. Characterization of the recombinant DGMA revealed that the highest maltogenic amylase activity occurred at $40^{\circ}C$ and pH 6.0. The half-life of catalytic activity at $65^{\circ}C$ and $55^{\circ}C$ were 8.2 min and 187.4 min, respectively. DGMA mainly hydrolyzed ${\beta}$-cyclodextrin, soluble starch, and pullulan and its efficient ratio of those substrates was 9:4.5:1.

The Effect of Annealing on sSEBS/Polyrotaxanes Electrolyte Membranes for Direct Methanol Fuel Cells

  • Won, Jong-Ok;Cho, Hyun-Dong;Kang, Yong-Soo
    • Macromolecular Research
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    • v.17 no.10
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    • pp.729-733
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    • 2009
  • Solution casting films of sulfonated poly[styrene-b-(ethylene-r-butylene)-b-styrene] copolymer (sSEBS)-based composite membranes that contained different amounts of organic, nanorod-shaped polyrotaxane were annealed at various temperatures for 1 h. The films' properties were characterized with respect to their use as polymer electrolyte membranes in direct methanol fuel cells (DMFCs). Different aspect ratios of polyrotaxane were prepared using the inclusion-complex reaction between $\alpha$-cyclodextrin and poly(ethylene glycol). The presence of the organic polyrotaxane inside the membrane changed the morphology during the membrane preparation and reduced the transport of methanol. The conductivity and methanol permeability of the composite membranes decreased with increasing polyrotaxane content, while the annealing temperature increased. All of the sSEBS-based, polyrotaxane composite membranes annealed at $140^{\circ}C$ showed a higher selectivity parameter, suggesting their potential usage for DMFCs.

Nafion Composite Membranes Containing Rod-Shaped Polyrotaxanes for Direct Methanol Fuel Cells

  • Cho Hyun-Dong;Won Jong-Ok;Ha Heung-Yong;Kang Yong-Soo
    • Macromolecular Research
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    • v.14 no.2
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    • pp.214-219
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    • 2006
  • Cast Nafion-based composite membranes containing different amounts of organic, nanorod-shaped polyrotaxane were prepared and characterized, with the aim of improving the properties of polymer electrolyte membranes for direct methanol fuel cell applications. Polyrotaxane was prepared using the inclusion-complex reaction between ${\alpha}$-cyclodextrin and poly(ethylene glycol) (PEG) of different molecular weights. The addition of polyrotaxane to Nafion changed the morphology and reduced the crystallinity. The conductivity of the composite membranes increased with increasing polyrotaxane content up to 5 wt%, but then decreased at higher polyrotaxane contents. Well-dispersed, organic polyrotaxane inside the membrane can provide a tortuous path for the transport of methanol, as the methanol permeability depends on the aspect ratio of polyrotaxane, which is controlled by the molecular weight of PEG. All of the Nafion-based, polyrotaxane composite membranes showed a higher selectivity parameter than the commercial Nafion films did.

A Data Base for Identification of Pseudomonas syringae pv. actinidiae, the Pathogen of Kiwifruit Bacterial Canker, Using Biolog Program (Biolog Program을 이용한 참다래 궤양병균 동정용 Data Base)

  • 고영진
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.125-128
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    • 1997
  • Reactions of Pseudomonas syringae pv. actinidiae to 95 carbon sources in a 96-well microplate (BiOLOG GN MicroPlateTM) were investigated. The bacterium used 9 carbon sources such as D-mannitol, sucrose, etc., but did not use 62 carbon sources such as $\alpha$-cyclodextrin, dextrin, etc. Based on the reactions, a user data base for identification of P. syringae pv. actinidiae was constructed in Biolog program (BiOLOG MicroLogTM 2 system). P. syringae pv. actinidiae isolates collected from kiwifruits could be identified automatically with high similarity using the user data base, which could diagnose rapidly and easily whether the tree was infected with bacterial canker or not.

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Amylolytic Enzymes Produced from Hyperthermophilic Archaebactorium Thermococcus profundus (고도 호열성 Archaebacterium Thermococcus profundus가 생산하는 Amylolytic Enzymes)

  • Jeong, Yeong-Cheol;Kim, Gyeong-Suk;No, Seung-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.7 no.4
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    • pp.259-266
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    • 1994
  • The hyperthermophilic archaebacterium Thermococcus profundus Isolated from a deep-sea hydrothermal vent system, produced several amylolytic enzymes such as extracellular amylase and pullulanase, intracellular a-1,4-91ucosidase in respone to the presence of complex carbohydrates In the growth medium. This strain showed high activities on 0.5% maltose than on complex carbohydrates One of the amylases was partially purified by ammonium sulfate precipitation, DEAE-Toyopearl chromatography. The amylase exhibited maximal activity at pH 5.5 and 80$^{\circ}C$, and was stable in the range of pH 5.5 to 9.5 and up to 80$^{\circ}C$ for 30 min. The enzyme activity was no dependence on Ca2+ and not inhibited by detergents. The amylase hydrolyzed soluble starch, amylose, amylopectin and glycogen to produce maltose and maltotriose with trace amounts of glucose, but not pullulan and ${\alpha}$-, ${\beta}$-, ${\gamma}$-cyclodextrin. Malto-oligosaccharides ranging from maltotetraose to maltoheptaose were hydrolyzed in an endo fashion.

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Purification and Characterization of $\beta$-Cyclodextrin Glucanotransferase Excreted by Bacillus firmus var. aikalophilus. (호알칼리성 Bacillus firmus가 생산하는 $\beta$-Cyclodextrin Glucanotransferase의 정제 및 효소반응 특성)

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.323-330
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    • 1998
  • Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0~9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

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Sensitivity Enhancement of Polydiacetylene Vesicles through Control of Particle Size and Polymerization Temperature (입자크기와 중합온도 제어를 통한 폴리다이아세틸렌의 센싱감도 향상)

  • Lee, Gil Sun;Oh, Jae Ho;Ahn, Dong June
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.400-404
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    • 2011
  • Many studies on polydiacetylene(PDA) have been investigated to apply to chemical and biological sensors due to their unique optical properties of color change from blue to red and fluorescence change from non-fluorescence to red fluorescence. Especially, high sensitivity against specific molecules is very important to apply polydiacetylenes to various sensors. In this study, we examined the effect of sensitivity enhancement of 10,12-pentacosadynoic acid(PCDA) vesicles in detection ${\alpha}$-cyclodextrin(CD) according to control of vesicle size by filters with different pore sizes and polymerization temperature. Colorimetric response(CR) was calculated using visible spectrometer. In order to investigate the effect of vesicle size on sensitivity of PDA vesicles, two PCDA vesicles were filtered without filtration and with 0.22 ${\mu}m$ filter. The two PCDA vesicles were polymerized at $25^{\circ}C$ and were incubated with ${\alpha}$-CD(5 mM) for 30 min. The CRs of the former and latter vesicles were 31.4% and 74.0%, respectively. Then, two PCDA vesicles filtered with 0.22 ${\mu}m$ filter were polymerized at $25^{\circ}C$ and $5^{\circ}C$ and were reacted with ${\alpha}$-CD(5 mM) for 30 min to examine the effect of polymerization temperature. The CRs of the former and latter vesicles were 74.0 and 99.2%, respectively. This suggests that vesicle sizes and polymerization temperature are key factors in enhancing the sensitivity of PDA vesicles. In addition, these results are expected to be useful to apply the PDA vesicles as biosensors to detect DNA, protein, and cells.

Saprolegnia ferax에 의한$\beta$-amylase의 생산 및 특성

  • Bai, Suk;Cho, Nam-Chul;Chun, Soon-Bai
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.109-114
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    • 1997
  • The Oomycete Saprolegnia ferax produces an extracellular $\beta$-amylase, Maximum enzyme yield was attained after 7 days of growth in YNB starch medium (pH 6.5) at 25$\circ$C. The amylase was pu- rified 24-fold by ultrafitration, HPLC DEAE column and HPLC gel filtration. The purfied enzyme was a monomeric glycoprotein with a molecular weight of about 44,000 dalton. The pH and temperature optima were 6.5 and 50$\circ$C, respectively. The enzyme was fairly stable up to 50$\circ$C and at acidic pH region (pH 4.0-7.0). The apparent Km and Vmax values of the enzyme against soluble starch were 0.77 mg/ml and 2,174 $\mu$moles/mg protein, respectively. Amino acid analysis indicated that the enzyme was enriched in alanine, glycine, leucine and acidic amino acid. Starch hydrolysis with the enzyme released maltose but not glucose, whereas maltotriose, Schardinger dextrin ($\alpha$-cyclodextrin) and pullulan were not hydrolysed by the enzyme. The enzyme was inhibited by Schardinger dextrin, p-chloromercuribenzoate(PCMB), CU$^{2+}$' and Hg$^{2+}$. Inhibition of the enzyme by PCMB could be reversed by the addition of cysteine and mercaptoethanol.

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FUNCTIONAL BEVERAGE FOR REDUCING BAD BREATH

  • Choi W;Kim S. R.;Kim Y. S;Park Y. K
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2001.12a
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    • pp.140-151
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    • 2001
  • This study was performed to examine a possible application of the beverage as a bad breath controlling food. To achieve this objective, methods of gas chromatography, electronic nose, sensory analysis and halimeter were used to detect reduction in odor intensities of bad breath caused by the functional beverage as well as its active ingredients. According to results of GC and electronic nose, adding green tea and champignon extracts to bad breath indicators, methylmercaptan and trimethylamine, resulted in significant reduction in headspace concentrations of two indicators. GC results revealed that headspace concentrations of 5 ug/ml of methylmercaptan and 30 ug/ml of trimethylamine added to various concentrations of two extracts were reduced up to $100\%$ after incubating mixtures at $37^{\circ}C$ for 5min. When the functional beverage was properly formulated with green tea extract, champignon extract and $\alpha$-cyclodextrin and evaluated for its deodorizing effect systematically, it also showed distinctive deodorizing activities against bad breath indicators. Conclusively, results obtained from this study might encourage introduction of a new type of bad breath control food in near future.

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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.