• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.63-69
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• 1991

Performance of column type bioreactor packed with immobilized cyclodextrin glucanotransferase (CGTase) on chitosan and Amberite IRA 900 was evaluated for cyclodextrin(CD) production. For CGTase immobilized on chitosan, the maximum CD conversion yield of 42% was achieved at the range of 88-168 units of immobilizied CGTase per gram of chitosan, retention time of 0.3 hr, and from 5.0% (w/v) of partially cyclized soluble starch. On the other hand, for CGTase immobilized on Amberite IRA 900, the maximum conversion yield of 40% was obtained at the range of 3.6-11.0 units of immobilized CGTase per gram of carrier and retention time of 1.2 hr from 5.0% of substrate. Above CD conversion yields are almost identical level with that can be obtained with soluble CGTase of 47%. The productivities of bioreactor packed with immobilized CGTase were 17.0g of CD/lㆍhr for amberite IRA 900 and 15.5g of CD/lㆍhr for chitosan. The partially cyclized starch with soluble CGTase were more suitable as substrate to achieve better CD conversion yield, and 5% (w/v) of partially cyclized soluble starch containing 10% (w/w) of CD was found to be most suitable to obtain maximum CD conversion yield.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.171-178
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• 1991

The two-stage enzyme reactor, packed with cyclodextrin glucanotransferase (CGTase) immobilized on Amberite IRA 900, coupled with ultrafiltration membrane was investigated for continuous production of cyclodextrin (CD). 5% (w/v) of soluble starch was partially cyclized, in the 0.1 l first-stage immobilized enzyme reactor, up to CD conversion yield of 10% (w/w) at retention time of 0.56hr and 1.5 units of immobilized CGTase/1g of carrier. In the second stage main immobilized enzyme reactor capacity of 1.5 l, the maximum CD conversion yield of 39% (w/v) was achieved at retention time of 2.8hr and 0.47 unit of CGTase/1 g of carrier. Unreacted residual dextrin was fractionated with ultrafiltration membrane, and then, recycled into the second-stage main bioreactor to increase the CD conversion yield. The most suitable membrane size and the volume concentration ratio (concentrate: filterate) for recycling of unreacted residual dextrin were found to be 5K dalton and 4:6, respectively. CD conversion yield was increased about 3~4% upon co-immobilization of pulluanase along with CGTase. Spent Amberite IRA 900 can be reutilized consecutively more than 3 times for immobilization of CGTase after regeneration.

• 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.

• Journal of Life Science
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• v.11 no.3
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• pp.230-234
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• 2001

The cyclodextrin glucanotransferase(CGTase) gene of Bacillus macerans was expressed in Saccharomyces cerevisiae and the recombinant CGTase was partially purified from the yeast culture supernatant. The optimal pH and temperature of the CGTase were found to be 6.0 and 5$0^{\circ}C$, respectively. The pH and temperature stabilities of the recombinant enzyme were significantly enhanced and the half life at 55$^{\circ}C$ was about 60 hr. When the recombinant CGTase was reacted with 5% soluble starch, the conversion yield of total cyclodextrin (CD) from starch was estimated to be 41% at 48 hr, whereas the wild type enzyme showed the yield of 12%. This improvement of conversion yield and thermal stability of CGTase may be useful for the development of low-cost CD production process.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.662-665
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• 2001

Enantioselective production of levofloxacin from ofloxacin butyl ester by porcine liver esterase was enhanced with the addition of ${\alpha}$-cyclodextrin(${\alpha}-CD$) and pH change. The conversion yield was increased from 27 to 64% and 100% with 150 mM substrate when the molar ratio of ${\alpha}-CD$ to substrate was 1 and 2, respectively. When 100 mM of substrate was added with the same molar ratio of ${\alpha}-CD$ at pH 5.6, the solubility was increased 3.8 times and the conversion yield was increased 4.4 times.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.163-170
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• 1991

Production of cyclodextrin (CD) directly from raw corn starch without liquefaction using cyclodextrin glucanotransferase (CGTase) was carried out in an agitated bead reaction system. Similar CD yield and production rate comparable with those of conventional method using liquefied starch were obtained. Especially high purity-CD in the reaction mixture without accumulation of malto-oligosaccharides was obtained. The maximum 54g/l of CD was obtained at raw starch concentration of 200g/l. CD yield was inversely proportional to raw starch concentration, and conversion yield was 0.48 at substrate concentration of 100g/l. The optimal amount of enzyme (CGTase unit/g raw starch) was found to be around 6.0. Granular structure of raw starch degraded by CGTase was observed by SEM in order to investigate the enhancing mechanism, along with those of acid or alkali pretreated raw starch, amylose, and amylopectin. Kinetic constants of CGTase on raw starch in an agitated bead reaction system were evaluated, and CGTase was competitively inhibited by CD.

• Microbiology and Biotechnology Letters
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• v.19 no.5
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• pp.514-520
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• 1991

Direct synthesis of cyclodextrin (CD) from extruded insoluble corn starch without liquefaction procedure using cyclodextrin glucanotransferase (CGTase) was carried out. Increased CD production rate and yield were achieved in heterogeneous enzyme reaction system containing extruded corn starch compared with those of conventional system employing liquefied or partially cyclized starch. At extruded starch concentration of 100 g/l the CD concentration and conversion yield were reached up to 54 g/l and 0.54, respectively. High purity of $\alpha \beta \gamma$-CDs without accumulation of undesirable malto-oligosaccharides was produced, furthermore, the residual extruded starch was easily separated by centrifugation from reaction mixture, whlch will facilitate the purification procedure. Granular structure of extruded starch was observed by SEM to investigate enzyme reaction mechanism. Supplemental addition of $\alpha$-amylase enhanced slightly the initial CD production rate, but it decomposed produced CD at the late stage. Various! extruded raw starches, such as, corn, rice, and barley were also suitable substrates for CD production.

• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.54-62
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• 1991

Cyclodextrin glucanotransferase(CGTase) derived from Bacillus macerans was immobilized by (1) covalent linkage on chitosan and chitin with glutaraldehyde, (2) adsorption on DEAE-cellulose and Amberite IRA 900 after succinylation, and (3) entrapment on alginate and polyacrylamide by cross linking. Adsorption on Amberite IRA 900 and covalent linking on chitosan were identified to be the most suitable immobilization methods considering the yield of activity and stability of immobilized CGTase. The enzymatic properties of immobilized CGTase were investigated and compared with those of the soluble CGTase. Thermal stability of CGTase immobilized on chitosan was increased from 50 to $55^{\circ}C$, and the optimum temperature of CGTase immobilized on Amberite IRA 900 was shifted from 55 to $50^{\circ}C$. The effect of molecular size of soluble starch (substrate) on immobilized CGTase investigated using partially liquefied substrates with different dextrose equivalent(DE). Cyclodextrin(CD) conversion yield augmented according to the increase of DE level for immobilized CGTase on Amberite IRA 900. CD conversion yield of partially cyclized starch with soluble CGTase was higher compared with liquefied one with ${\alpha}-amylase$.

• Biotechnology and Bioprocess Engineering:BBE
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• v.4 no.1
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• pp.32-35
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• 1999

Addition of ${\beta}$-cyclodextrin (${\beta}$-CD) polymer during the biotransformation of digitoxin into digoxin using cell suspension cultures of Digitalis lanata enhanced the conversion yield. Digitoxin showed better adsorption to CD polymer compared to digoxin, so that the optimization of addition time was found to be necessary. In the case of adding CD polymer 24 hours after the feeding of substrate digitoxin, the highest digoxin production could be achieved. At this period, digitoxin was almost consumed by cells and productivity was proportionally enhanced according as the amount of substrate was increased. Immobilization of CD polymer did not promote the biotransformation. When 3.33 g/L of CD selective inclusion complex formation could be expected. Adsorption rate was found to be rapid and saturation was obtained within 10 hours of contact.

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.293-297
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• 2002

The cyclodextrin glucanotransferase (CGTase) gene from Bacillus stearothermophilus NO2 was expressed in Saccharomyces cerevisiae 2805 under the adhl promoter. The CGTase was purified from S. cerevisiae 2805/pVT-CGTS. The purified enzyme exhibited a optima of activity around pH 7.0 and $65^{\circ}C$. Thermal stability of the enzyme was increased fairly as compared with the CGTase of B. stearothermophilus NO2. The conversion yield of cyclodextrin (CD) and the production ratio of $\alpha$-, $\beta$,-, ${\gamma}$-CD from starch were showed similarly aspect to the CGTase of B. stearothermophilus NO2.