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

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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.

Development of Functionality in Cheese (기능성 향상 치즈 개발 연구)

  • Ahn, Sung-Il;Choi, Kyung-Hoon;Kwak, Hae-Soo
    • Journal of Dairy Science and Biotechnology
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    • v.29 no.1
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    • pp.65-73
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    • 2011
  • Cheese is a nutritious food with various balanced nutrients, such as proteins, peptides, amino acids, fats, fatty acids, vitamins and minerals. Domestic cheese varieties and quality need to be improved to prevent imported cheese. To develop those cheeses, search for previous works and research for new products are needed. In cheese ripening of hard cheese, such as Cheddar or Parmesan cheese, is ripened for 2 to 24 months at 2 to 16$^{\circ}C$ to develop desired cheese flavor and body characteristics. Long time with low temperature to ripen the cheese requires high expenses. So accelerated cheese ripening is a good potential for saving in industry. Methods for acceleration of cheese ripening are temperature control, addition of bacteria or enzymes. To develop the functionality of cheese, addition of microencapsulated various probiotics and nutrients, such as iron, removal of cholesterol by crosslinked ${\beta}$-cyclodextrin, lowering blood cholesterol and serum glucose by nanopowdered functional materials et al. are necessary. Therefore, this review focused on the functionality of cheese, such as the acceleration of cheese ripening, microencapsulated probiotics and iron, and cholesterol removal.

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Molecular Cloning and Functional Expression of esf Gene Encoding Enantioselective Lipase from Serratia marcescens ES-2 for Kinetic Resolution of Optically Active (S)-Flurbiprofen

  • Lee, Kwang-Woo;Bae, Hyun-Ae;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.74-80
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    • 2007
  • An enantioselective lipase gene (esf) for the kinetic resolution of optically active (S)-flurbiprofen was cloned from the new strain Serratia marcescens ES-2. The esf gene was composed of a 1,845-bp open reading frame encoding 614 amino acid residues with a calculated molecular mass of 64,978 Da. The lipase expressed in E. coli was purified by a three-step procedure, and it showed preferential substrate specificity toward the medium-chain-length fatty acids. The esf gene encoding the enantioselective lipase was reintroduced into the parent strain S. marcescens ES-2 for secretory overexpression. The transformant S. marcescens BESF secreted up to 217kU/ml of the enantioselective lipase, about 54-fold more than the parent strain, after supplementing 3.0% Triton X-207. The kinetic resolution of (S)-flurbiprofen was carried out even at an extremely high (R,S)-flurbiprofen ethyl ester [(R,S)-FEE] concentration of 500 mM, 130 kU of the S. marcescens ES-2 lipase per mmol of (R,S)-FEE, and 1,000 mM of succinyl ${\beta}-cyclodextrin$ as the dispenser at $37^{\circ}C$ for 12h, achieving the high enantiomeric excess and conversion yield of 98% and 48%, respectively.

Formulation and Evaluation of Moisture-activated Acyclovir Patches (수분 감응성 아시클로버 패취제의 설계 및 평가)

  • Kim, Ah-Mee;Gwak, Hye-Sun;Chun, In-Koo
    • Journal of Pharmaceutical Investigation
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    • v.36 no.6
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    • pp.393-399
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    • 2006
  • This study was aimed to design, formulate and characterize the moisture-activated patches containing acyclovir for antiviral action. Gel intermediates for film-type patches were prepared with mucoadhesive polymer, viscosity builders, enhancers and acyclovir. Patches containing acyclovir were characterized by in vitro measurement of drug release rates through a cellulose barrier membrane, and of drug flux through the hairless mouse skin. Film-type patches obtained were uniform in the thickness and showed a mucoadhesive property when contacted with moisture. The formulation was optimized, which consisted of $Cantrez^{\circledR}$ AN-169(2%), $Kollidon^{\circledR}$ VA 64(1%), $Natrosol^{\circledR}$(1%), hydroxypropyl-$\beta$-cyclodextrin(1%) and dimethylsulfoxide(0.5%). Release rates of acyclovir patches increased dose-dependently. The addition of terpenes such as d-limonene or cineole increased release rates of acyclovir, but decreased permeation rates. The permeation rates were enhanced by 2 and 2.5 times by the addition of glycyrrhizic acid ammonium salt and sodium glycocholate, respectively, compared with that of no enhancer. These results suggest that it may be feasible to deliver acyclovir through the skin or gingival mucosa from the moisture-activated patches.

UV-HPLC Determination of Carbowyl Group Using 2-Bromoacetyltriphenylene as a Pre-labeling Reagent - The isolative determination of prostaglandin $E_2$ and $F_2{\alpha}$ by HPLC (2-Bromoacetyltriphenylene 유도체화제를 이용한 카르복실기 함유성분의 분석법 (I) - 프로스타글란딘 $E_2$$F_2{\alpha}$ 혼합물의 HPLC에 의한 분리정량)

  • 이왕규;정해수;김박광
    • YAKHAK HOEJI
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    • v.30 no.6
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    • pp.311-316
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    • 1986
  • A new UV labeling reagent was developed and used in HPLC for the determination of prostaglandin $E_2$ which have weak UV light-absorbing property. This reagent, 2-bromoacetyltriphenylene, was synthesized by the bromination of 2-acetyltriphenylene which was obtained from triphenylene by Friedel-Crafts reaction. The wave length maximum (${\lambda}_{max}^{CH_3CN}$ of this reagent was 268nm. Prostaglandin E$_2$ was extracted from prostaglandin E$_2$-$\beta$-cyclodextrin using a Sep-pak $C_{18}$ cartridge. The prostaglandin E$_2$ was labeled with 2-bromoacetyl-triphenylene in aectonitrite using 18-crown-6-ether as catalyst. Derivatized prostaglandins were separated on a reversed-phase column (Radial-pak) $\mu$-Bondapak $C_{18}$ using acetonitrile: water=60:40 as mobile phase. The effluent was monitored by UV detector at 254nm filter kit. Linearity of calibration curve was obtained between 30ng and 140ng, and the lower limit of detection was 5ng.

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Effects of nonionic surfactants on the partitioning of naphthalene in soil/water system (토양과 수용액상에서 나프탈렌의 분배에 관한 비이온성 계면활성제의 영향)

  • 하동현;고석오;신원식;김영훈;전영웅;송동익
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2002.09a
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    • pp.283-286
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    • 2002
  • Solubilization and partitioning of naphthalene was investigated in an aqueous system containing soils and surfactants. The environmental behavior of polycyclic aromatic hydrocarbons(PAHs) was mainly governed by their solubility and partitioning properties on soil media in a subsurface system. In surfactant-enhanced remediation systems, surfactants might be an additional variable. a natural soil ,silica and kaolinite were tested as soil media. two nonionic surfactants, Triton X-100 and Hydropropy1-$\beta$-cyclodextrin (HPCD) were employed for naphthalene solubilization. Naphthalene showed linear on natural soil while non-linear sorption on silica and kaolinite. Soils have higher sorption capacity for Triton X-100 than HPCD indicating Triton X-100 formed ad-micelle on the soil surface. Desorption study showed a hysterysis and reversible desorption. The partitioning coefficient(K$_{D}$) of naphthalene was increased as the concentration of surfactant was increased. (below CMC), however, the coefficient was decreased above CMC. This indicates that naphthalene is partitioned into the micelles and the partition occurs competitively on both ad-micelle and free micelles as surfactant concentration increases. Therefore, the target compounds to be dissolved into aqueous phase in a surfactant enhanced remediation system might be highly partitioned on to the ad-micelle resulting in an adverse effect rather increased solubilization would be achieved.d.

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Optimization of Pine Flavor Microencapsulation by Spray Drying

  • Lee, Shin-Jo;Lee, Yang-Bong;Hong, Ji-Hyang;Chung, Jong-Hoon;Kim, Suk-Shin;Lee, Won-Jong;Yoon, Jung-Ro
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.747-751
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    • 2005
  • Microencapsulation of pine flavors was investigated to determine the optimum wall material and spray drying condition. ${\beta}$-Cyclodextrin, maltodextrin, and a 3:1 mixture of maltodextrin and gum arabic were evaluated as wall materials. The latter mixture was determined to be the best wall material based on dispersion capacity and flavor yield. Spray drying effectiveness was evaluated using a $3^3$ fraction factorial design and statistical analysis. The optimum operation condition was an inlet air temperature of $175^{\circ}C$, inlet airflow rate of $0.65\;m^3/min$ and atomizing pressure of 180 kPa, which resulted in a 93% flavor yield. The best particle shape observed by SEM was a round globular shape obtained under the above spray drying condition, whereas lower temperatures and higher inlet airflow rates resulted in initial and full collapses, respectively. The round globular shapes remained stable for at least one month.

Synthesis of an Amphiphilic Poly(${\gamma}$-Glutamic Acid)-Cholesterol Conjugate and Its Application as an Artificial Chaperone

  • Lee, Eun-Hye;Kamigaito, Yoshiki;Tsujimoto, Takashi;Uyama, Hiroshi;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1424-1429
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    • 2010
  • A poly(${\gamma}$-glutamic acid) (${\gamma}$PGA)-cholesterol conjugate was synthesized and its properties were then evaluated. The conjugate exhibited an amphiphilic nature derived from the hydrophilic ${\gamma}$PGA backbone and the hydrophobic cholesterol side chain. The conjugate spontaneously formed nanoparticles, becoming an aqueous solution when at low concentrations, and at high concentrations the result was the formation of a physical gel. By utilizing the self-aggregating properties of the conjugate in water, an artificial chaperone was developed. A complex of protein, with the nanoparticles of the conjugate, was formed and the protein was released upon the dissociation of the nanoparticles through the addition of ${\beta}$-cyclodextrin. For denatured carbonic anhydrase, the activity was recovered in the artificial chaperone of the nanoparticle conjugate.

Physicochemical Stability of Leucine Enkephalin and $[D-Ala^2]$-Leucine Enkephalinamide in Buffered Aqueous Solution (완충 수용액중 로이신엔케팔린 및 [D-알라$^2]$-로이신엔케팔린아미드의 물리화학적 안정성)

  • Park, In-Sook;Chun, In-Koo
    • YAKHAK HOEJI
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    • v.38 no.5
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    • pp.488-495
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    • 1994
  • To evaluate the feasibility of transmucosal delivery of leucine enkephalin (Leu-Enk) and its synthetic analog, $[D-Ala^2]$-leucine enkephalinamide (YAGFL), their physicochemical stabilities in aqueous buffered solutions were first investigated using a stability indicating high performance liquid chromatography. The degradation of Leu-Enk and YAGFL followed the pseudo-first-order kinetics. From the pH-rate profiles, it was found that the maximal stability of the two pentapeptides was at the pH of about 5.0. The shelf lives $(t_{90%})$ for the degradation of Leu-Enk and YAGFL at pH 5.0 and $40^{\circ}C$ were found to be 48.13 and 50.9 days, respectively. From the temperature dependence of the degradation, activation energies for Leu-Enk and YAGFL were calculated to be 13.61 and 13.47 kcal/mole, respectively. A higher ionic strength and a higher initial peptide concentration in buffered solution slowed the degradation of the two pentapeptides. The addition of 2-hydroxypropyl-${\beta}$-cyclodextrin into the peptide solution did not affect the stability significantly.

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Use of Biosurfactant for the Removal of Organic Pollutants in Soil/Groundwater (바이오 계면활성제에 의한 토양/지하수내 유기성 오염물질 제거)

  • Ko, Seok-Oh;Yoon, Seok-Pyo
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.2
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    • pp.193-201
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    • 2000
  • Partitioning of hydrophobic organic compounds (HOCs) to a biosurfactant, hydroxypropyl-${\beta}$-cyclodextrin (HPCD), was conducted to evaluate the feasibility of using HPCD to remove HOCs from soil/groundwater. HOC partitioning to HPCD was very fast, with over 95% of the complexation occurring within 10 min. Some influence of solution chemistry and HOC concentration on HOC-HPCD complex formation coefficients was observed. HPCD sorption on soil as quantified by both a fluorescence technique and total organic carbon measurements was negligible, indicating no significant affinity of HPCD for the solid phase. Although the HOC solubilization capability of HPCD was lower than that of synthetic surfactants such as SDS and Tween 80, HPCD can be effective in removing sorbed HOCs from a model subsurface environment, primarily because of its negligible sorption to the solid phase (i.e., all the HPCD added facilitates HOC elution). However, in contrast with conventional surfactants, HPCD becomes relatively less effective for HOC partitioning with increasing HOC size and hydrophobicity. Therefore, comparisons between HPCD and synthetic surfactants for enhanced remediation applications must consider the specific HOC(s) present and the potential for surfactant material losses to the solid phase, as well as other more generally recognized considerations such as material costs and potential toxicological effects.

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