• Analytical Science and Technology
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• v.13 no.5
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• pp.549-557
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• 2000

FRIR-ATR spectrometry has been used to monitor the aqueous reactions of compounds without distinct chromophores in ultraviolet and visible regions. For example, hydrolysis reactions of ${\alpha}$-cyclodextrin and ${\gamma}$-cyclodextrin in acidic aqueous solution were studied. FTIR-ATR method has been used for the monitoring of cyclodextrin hydrolysis in 1.0 M. 0.5 M, and 0.1 M HCl solutions, respectively. We also found that the hydrolysis of ${\alpha}$-cyclodextrin produced glucose, but the hydrolysis of ${\gamma}$-cyclodextrin proceeded further to give more fragmented products than glucose.

• Applied Biological Chemistry
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• v.33 no.1
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• pp.79-86
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• 1990

Cyclodextrin hydrolase from Bacillus stearothermophilus KFCC 21203 was purified and the properties of the purified enzyme were investigated. The enzyme was purified 15 folds with 77 % recovery by ammonium sulfate fractionation, DEAE-cellulose chromatography, and Ultro AcA 34 gel filtration. The specific activity and the molecular weight of the enzyme were 1.30 units/mg protein and about 29,500, respectively, The maximum activity of the enzyme was shown at $55^{\circ}C$ and pH 5.5. However, stable temperature and pH were $40^{\circ}C$ and $5.0{\sim}8.0$, respectively. The Km value for ${\gamma}-cyclodextrin$ was $3.78{\times}10^{-3}$ M. The degradation activity of the enzyme was selectively high for ${\gamma}-cyclodextrin$, and very low for ${\beta}-cyclodextrin$, but not for ${\alpha}-cyclodextrin$. The decomposed products of ${\gamma}-cyclodextrin$ were mainly glucose and maltose, and a little mlatotriose. The activity of the enzyme was very high for amylose, potato starch, corn starch, amylopectin and maltooligomer, and relatively high for glycogen and dextrin. The decomposed products of them were mainly glucose and maltose.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.156-163
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• 1992

Bacillus sp. was isolated from soil for its strong activity of cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19). The enzyme was purified by gel filtration and anion exchange column chromatography using FPLC. The purified enzyme exhibited its maximum CGTase activity in the pH range of 6~8 and the temperature range of 50~$70^{\circ}C$. The molecular weight was estimated as 114,000 by SDS-PAGE. The isoelectric point of the enzyme was 4.3. The CGTase of Bacillus sp. E l produced $\beta$-cyclodextrin mainly and did not produce a-cyclodextrin. The product ratio of $\beta$-cyclodextrin to $\gamma$-cyclodextrin was 7:l.

• YAKHAK HOEJI
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• v.39 no.2
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• pp.175-184
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• 1995

The interaction of omeprazole(OMP) with $\gamma$-cyclodextrin($\gamma$-CyD) was investigated by solubility study and the complexation was confirmed by means of UV/VIS spectrophotometer, circular dichroism, differential scanning calorimeter, and $^{1}$H nuclear magnetic resonance spectra. The stability, dissolution rate, and partition coefficient of the complex were measured. The results present that the benzimidazole moiety and a part of pyridine ring containing sulfur atom of OMP might be included into the cavity of $\gamma$-CyD and the formation type of inclusion complex appeared to be B$_{s}$. The stoichiometric ratio of OMP to $\gamma$-CyD in the complex was found to be 1:1 and the stability constant of the complex found to be 97.1 M$^{-1}$. And the dissolution rate of OMP was markedly increased by inclusion complex formation with $\gamma$-CyD, and so it was above 90% in 5 min. from solid complex. Oil to water partition coefficient of OMP-$\gamma$-CyD complex was 60, which is significantly higher than that of OMP itself, 36.4. The degradation rate constant of OMP were greater than OMP-$\gamma$-CyD complex in aqueous solutions of various pHs, and the half lives of OMP and OMP-$\gamma$-CyD at pH 9 were 279.2 and 509.9 days, respectively, showing that the complex was more stable than OMP, therefore it was thought that OMP was stabilized by inclusion formation with $\gamma$-CyD.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.465-471
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• 1997

The cyclodextrin glycosyltransferase (CGTase, EC 3.2.1.19) from Bacillus brevis CD162 was purified by precipitating with ammonium sulfate, DEAE-Sepharose CL-6B column chromatography and Sephadex G-150 column chromatography. The molecular mass and pI of the purified enzyme were estimated to be 74,000 and 6.3 by SDS-PAGE and isoelectric focusing, respectively. The purified enzyme was clearly identified as the CGTase by zymogram after SDS-PAGE. The optimum pH and temperature for the enzyme activity were 8.0 and $55^{\circ}C$, respectively. The enzyme was stable at the range of pH $5.5{\sim}9.0$, and up to $50^{\circ}C$. The amino acid sequence from the $NH_2-terminal$ of the purified CGTase was Ser-Val-Thr-Asn-Lys-Val-Asn-Tyr-Ser-Lys-Asp-Val-Ile-Tyr-Gln. The yields of the products from starch as the substrate were 1.3% for ${\alpha}-$, 33.9% for ${\beta}-$, and 9.7% for ${\gamma}-cyclodextrin$.

• Applied Biological Chemistry
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• v.38 no.1
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• pp.42-48
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• 1995

The experiment was carried out to purify and to investigate the properties of the cyclodextrinase produced from Bacillus megaterium KFCC 11855. The enzyme was partially purified with $(NH_4)_2SO_4$ and chromatography on DEAE-trisacryl, Ultrogel AcA 34, DEAE-trisacryl and Ultrogel HA. The optimum temperature and pH of the purified enzyme were $60^{\circ}C$ and 6.0, respectively. The enzyme was stable at the temperature of $45^{\circ}C$ below and at the pH range of $6.0{\sim}9.0$, respectively. The Km value for ${\gamma}-cyclodextrin$ was 0.903 mM. The enzyme activity was increased by $Mg^{2+}$ and $Mn^{2+}$, but decreased by $Hg^{2+}$ and $Cu^{2+}$. The enzyme degraded ${\gamma}-cyclodextrin$ but not ${\alpha}-cyclodextrin$. The degree of ${\beta}-CD$ degradation by the enzyme was very low. The decomposed products of ${\gamma}-cyclodextrin$ by the enzyme were mainly glucose, maltose and a little amount of maltotriose.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.2
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• pp.351-357
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• 1997

The cyclodextrin glycosyltransferase(EC 3.2.1.19) from Bacillus firmus was purified by precipitating with ammonium sulfate followed by, DEAE-Sephadex A-50 column chromatography and Sephadex G-100 column chromatography. In this way, we were able to obtain the single band protein on SDS-PAGE with a yield of 12%, whose purity was 49 fold. The purified CGTase was identified as a protein having molecular weight of approximately 80,000 dalton and isoelectric point of 9.6. The optimum pH and temperature for the enzyme activity were 8.0 and $65^{\circ}C$, respectively. The enzyme was stable at between pH 5.5 and 9.0 and up to $50^{\circ}C$. After 24hr of enzyme reaction using soluble starch as substrate, the ratio of ${\alpha}-$, ${\beta}-$ and ${\gamma}-cyclodextrin$ production was 0.01 : 2.90 : 1.00, respectively. And this CGTase pro-duced mainly ${\beta}-$ and ${\gamma}-cyclodextrin$.

• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.444-449
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• 1991

Synthesis of maltosyl-$\beta$-cyclodextrin using maltose ($G_2$) and $\beta$-cyclodextrin ($\beta$-CD) as substrates through the reverse reaction of pullulanase was investigated. The optimal conditions for the condensation reaction were as below: mixing ratio of maltose to $\beta$-CD of 12.7, mixed substrate concentration of 70% (w/w, 70 g/100 ml $H_2O$), and amount of pullulanse of 350 units/100 ml. The concentration of synthesized maltosyl-P-CD concentration was reached up to 2.31 g/100 rnl at above reaction conditions, which corresponded the conversion yield of 43% (w/w, g of branched-CD/g of CD). The synthesis of maltosyl-$\alpha >\gamma >\beta$-CD was also attempted, and conversion yield was in the order of a>y>J3-CDs. Condensation reaction between various maltooligosaccharides ($G-1\sim G_6$ showed that maltose was the most effective oligorner for condensation reaction with $\beta$-CD. To increase the conversion yield various alcohols were added into the reaction mixture, amyl alcohol was found to be the most acceptable alcohol for increasement of convesion yield which increased from 43.0 to 83.0% upon addition of same volume of amyl alcohol into the reaction mixture.

• Korean Journal of Food Science and Technology
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• v.26 no.1
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• pp.6-11
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• 1994

A cyclodextrin glucanotransferase(CGTase)-producing Bacillus sp. I-5 was isolated from soil and the enzyme exhibited the maximum reaction rate at pH 8.0 and $50^{\circ}C$. It was found that CGTase of I-5 produced ${\beta}-$ and ${\gamma}-CD$ mainly but the production ratio of cyclodextrins (CDs) was influenced by the buffer solution. Sodium acetate significantly stimulated the formation of ${\gamma}-CD$, increasing the content by 35%. The production of CDs was influenced by DE value of starch. The results indicated that DE value in the range of $3.5{\sim}6.0$ were most effective for the CD formation. CGTase was immobilized on the reversibly soluble-insoluble carrier, hydroxypropyl mothylcellulose acetate succinate. The immobilized CGTase was soluble at pH 7.5, and precipitated easily at pH 6.0. Enzyme reactor was designed to produce CD continuously. It was composed of three major stages-CD produttion by immobilized CGTase, conversion of the residual dextrin to glucose by amylase and glucoamylase and alcohol fermentation by yeasts to remove the glucose into alcohol. The yield of total CDs was 3.65g from 10g soluble starch.

• KSBB Journal
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• v.13 no.1
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• pp.26-30
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• 1998

The suspension cultures of Mentha piperita produce menthol which has very low solubility in water due to its hydrophobicity. This can be considered as a factor for its low production in the suspension suspension cultures. Cyclodextrin has the hydrophobic cavity inside the molecule in which menthol can be captured and allow to form a stable complex. The suspension culture of Mentha piperita showed 70% higher production enhancement in the medium containing 1.5%(w/v) $\beta$-cyclodextrin than the control. $\beta$-cyclodextrin had no adverse effect on the cell growth and showed the best result among $\alpha$-, $\beta$- and $\gamma$-cyclodextrins tested in terms of menthol production. We demonstrated that $\beta$-cyclodextrin can be used to enhance the production of menthol in the suspension cultures by capturing hydrophobic menthol into the cavity of cyclodextrin molecules.