• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.725-729
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• 2008

An enzyme reactor installed with ultrafiltration membrane was developed to produce ${\alpha}-,\;{\beta}-$, and ${\gamma}$-cyclodextrins (CDs) from soluble starch by Bacillus macerans cyclodextrin glycosyltransferase (CGTase) tagged with 10 lysines at its C-terminus (CGTKIOase). Ultrafiltration membrane YM10 with 10,000 of molecular cutoff was chosen for membrane modification and CD production. A repeated-batch type of the enzyme reaction with free CGTK10ase resulted in a ${\alpha}$-CD yield of 24.0 (${\pm}1.5$)% and a productivity of 4.68 (${\pm}0.88$) g/l-h, which were 7 times higher that those for CGTK10ase immobilized on modified YM10 membrane. Addition of 1-nonanol increased CD yields by 30% relative to the control, which might be due to prevention of the reversible hydrolysis of CDs.

• KSBB Journal
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• v.10 no.2
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• pp.149-158
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• 1995

The capacity of inclusion complex formation between ${\alpha}$-, ${\beta}$-, ${\gamma}$-cyclodextrins(CDs) and various compounds, such as pH indicators, biloslalns, glycoside, amino acid, and fatty acids, was compared. Fatty acid was identified as the most suitable ligand for fractionation of CDs in terms of capacity and selectivity. The effects of complex formation conditions, such as, mixing ratio of CD and fatty acid, pH, ionic strength, and temperature, on the capacity of fatty acrid-CD complex was also investigated. The carbon number of fatty acids was identified as the most significant factor determining the capacity and selectivity of inclusion complex formation of CDs. Capric acid(C10) and palmitic acid(C16) showed high specificity for ${\alpha}$- and ${\beta}$-CDs, respectively. Under the optimal conditions, the molar ratio of complex formed was found to be 1.0:2.6 for ${\alpha}$-CD/capric acid and 1.0:1.9 for ${\beta}$-CD/palmitic acid. X-ray diffraction and infrared spectrum of the formed inclusion complex were analyzed. The changes of enthalpy($\Delta$H) of the inclusion complex formation reaction was evaluated by differential scanning calorimetry, showed that the reaction was endothermic.

• 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.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.792-799
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• 2007

A gene encoding a putative glycogen-debranching enzyme in Sulfolobus shibatae(abbreviated as SSGDE) was cloned and expressed in Escherichia coli. The recombinant enzyme was purified to homogeneity by heat treatment and Ni-NTA affinity chromatography. The recombinant SSGDE was extremely thermostable, with an optimal temperature at $85^{\circ}C$. The enzyme had an optimum pH of 5.5 and was highly stable from pH 4.5 to 6.5. The substrate specificity of SSGDE suggested that it possesses characteristics of both amylo-1,6-glucosidase and $\alpha$-1,4-glucanotransferase. SSGDE clearly hydrolyzed pullulan to maltotriose, and $6-O-\alpha-maltosyl-\beta-cyclodextrin(G2-\beta-CD)$ to maltose and $\beta$-cyclodextrin. At the same time, SSGDE transferred maltooligosyl residues to the maltooligosaccharides employed, and maltosyl residues to $G2-\beta-CD$. The enzyme preferentially hydrolyzed amylopectin, followed in a decreasing order by glycogen, pullulan, and amylose. Therefore, the present results suggest that the glycogen-debranching enzyme from S. shibatae may have industrial application for the efficient debranching and modification of starch to dextrins at a high temperature.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.9
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• pp.1468-1472
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• 2007

The present study was carried out to determine the optimum conditions of different factors (ratio of lard to water, ${\beta}$-CD concentrations, mixing temperature, mixing time and mixing speed) on cholesterol reduction from lard by using crosslinked ${\beta}$-CD. Crosslinked ${\beta}$-CD was prepared with adipic acid. When the lard was treated under different conditions, the range of cholesterol removal was 91.2 to 93.0% with 5% crosslinked ${\beta}$-CD, which was not significantly different among treatments. In a recycling study, cholesterol removal with crosslinked ${\beta}$-CD in the first trial was 92.1%, which was similar to that with new crosslinked ${\beta}$-CD. In up to eight time trials, over 90% of cholesterol removal was found. The present study indicated that the optimum conditions for cholesterol removal using crosslinked ${\beta}$-CD were a 1:3 ratio of lard to water, 5% crosslinked ${\beta}$-CD concentration, $40^{\circ}C$ mixing temperature, 1 h mixing time and 150 rpm mixing speed. In addition, crosslinked ${\beta}$-CD made by adipic acid resulted in an effective recycling efficiency.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.495-500
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• 1997

To isolate a gene for cyclodextrin glycosyltransferase (CGTase) from alkalophilic Bacillus sp. E1, polymerase chain reaction (PCR) amplification was carried out. Direct molecular cloning of 1.2 kbp fragment and partial nucleotide sequence analysis of the PCR amplified clone, pH12, showed close homology with CGTases from Bacillus species. To investigate the genomic structure of the gene, Southern blot analysis of genomic DNA was carried out with the clone pH12 as a molecular probe. It showed that 5.3 kbp XbaI fragment was hybridized with the probe pH12. To isolate a genomic clone, genomic DNA library was constructed and a genomic clone for CGTase, pCGTE1, was isolated. Nucleotide sequence analysis of the clone pCGTE1 revealed that BCGTE1 contained 2,109 bp open reading frame encoding a polypeptide of 703 amino acids and showed over 94.3% amino acid sequence homology with CGTase of ${\beta}-cyclodextrin$ producer, Bacillus sp. KC201.(Received October 7, 1997; accepted October 20, 1997)

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.10a
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• pp.358-362
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• 2004

본 연구에서는 가교화 ${\beta}-CD$로 크림치즈의 콜레스테롤을 효과적으로 제거하는 실험을 수행 하였으며, 크림치즈의 이화학적 변화와 관능적 특성을 살펴보았다. 크림치즈를 만들기 위한 유지방 함량 36%의 크림을 가교화 ${\beta}-CD$로 처리 시 콜레스테롤 제거 최적 조건은 가교화 ${\beta}-CD$ 10%를 첨가해 교반온도 $20^{\circ}C$, 교반시간 30분, 교반속도 800rpm으로 실험한 결과 콜레스테롤 제거율이 평균 82.0%로 나타났다. 가교화 ${\beta}-CD$처리 크림치즈는 short-chain fatty acid의 경우 저장 기간이 지남에 따라 control과 powder ${\beta}-CD$ 처리한 크림치즈에 비해 저급 지방산 생성에 변화가 거의 없고, 쓴맛을 내는 아미노산의 경우 저장 기간 동안 control과 powder ${\beta}-CD$처리 크림치즈에 비해 생산량이 현저하게 적었다. 또한 가교화 ${\beta}-CD$처리 크림치즈의 조직검사에서 다른 항목에서보다 저장 기간 동안 응집성이 변함이 없으며 그 수치가 높게 평가되었다. 관능검사 결과, 가교화 ${\beta}-CD$처리 크림치즈는 저장 기간 동안 쓴맛의 증가가 거의 없었고 전체적인 기호도 또한 높았다. 위 실험 결과에 따르면, 가교화 ${\beta}-CD$ 사용결과 cholesterol 제거율이 높으며, 제품에 적용시 재활용이 가능하고 품질이 향상되므로 이를 유가공 산업에 활용이 가능할 것으로 기대된다.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.05a
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• pp.317-321
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• 2005

본 연구결과, 콜레스테롤 제거 크림치즈를 만들기 위한 유지방 함량 36%의 크림의 콜레스테롤 제거 최적 조건은 adipic acid로 가교시킨 ${\beta}-CD$ 첨가량 10%, 교반온도 $20^{\circ}C$, 교반시간 30분, 교반속도 800rpm 이었으며, 이 때 콜레스테롤 제거율은 평균 91.42%로 나타났다. 가교화 ${\beta}-CD$ 처리 크림치즈의 관능검사 결과, 가교화 ${\beta}-CD$ 처리 크림치즈는 control과 powder ${\beta}-CD$ 처리한 크림치즈에 비해 저장 기간 동안 쓴맛의 증가가 거의 없었고, 빵에 발리는 정도도 매우 우수하였으며, 전체적인 기호도 또한 높았다. short-chain fatty acid의 경우 저장 기간이 지남에 따라 가교화 ${\beta}-CD$ 처리 크림치즈는 control과 powder ${\beta}-CD$ 처리한 크림치즈에 비해 저급 지방산 생성에 변화가 거의 없었다. 조직검사에서 가교화 ${\beta}-CD$ 처리 크림치즈는 저장 기간 동안 control에 비해 응집성, 탄력성, 점착성의 변화가 없으며 그 수치가 높게 평가되었다. 또한 가교화 ${\beta}-CD$의 재활용률은 97.82%로 매우 높게 나타났다. 위 실험 결과에 따르면, 가교화 ${\beta}-CD$는 콜레스테롤 제거율이 높으며, 제품에 적용 시 품질이 향상될 뿐만 아니라, 재활용률도 높아 이를 유가공 산업에 다양하게 활용이 가능할 것으로 기대된다.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.305-305
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• 2006

The molecular structure and dynamics of inclusion compounds (ICs) consisting of n-perfluoroalkane (PFA) guests and ${\Box}-cyclodextrin$ (${\Box}-CD$) host were investigated using $^{19}F$ magic angle spinning (MAS) and $^{1}H{\to}^{19}F$ cross polarization (CP) / MAS NMR spectroscopy with the aid of thermal analyses, FT-IR spectroscopy, X-ray diffraction, and $^{1}H{\to}^{19}F$ CP/MAS technique revealed that $C_{9}F_{20}$ molecules included in ${\Box}-CD$ undergo vigorous molecular motion and partly come out of the ${\Box}-CD$ channel above $80^{\circ}C$. In case of $C_{20}F_{42}/{\Box}-CD$, an exothermic peak is observed by differential scanning calorimetry (DSC) at ca. $40^{\circ}C$ which suggests that ${\Box}-CD$ molecules become mobile and commence rearrangements that form more ordered structures at higher temperatures.

• Journal of the Korean Society of Tobacco Science
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• v.24 no.2
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• pp.94-100
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• 2002

Cyclodextrin glucanotransferase from Bacillus stearothemophilus and Bacillus macerans synthesized maltol and ethyl maltol monoglucoside, with a series of its maltooligo-glucosides by transglycosylation with dextrin as a donor, and maltol or ethyl maltol as an acceptor. The monoglucoside formed from reaction mixture of maltol or ethyl maltol by the successive actions of Bacillus stearothemophilus cyclodextrin glucanotransferase and Rhizopus glucoamylase was isolated by Diaion HP-20 column and silica gel column chromatography. The structure of the isolated monoglucoside was identified as maltol-$\alpha$-D-glucoside and ethyl maltol-$\alpha$-D-glucoside, respectively, by FAB-MS, UV, $^1$H-NMR, $^{13}$ C-NMR spectra and products by hydrolysis with acid, $\alpha$ - and $\beta$ -glucosidases.