• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1201-1205
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• 2009

Corn starches with different amylose contents were enzymatically modified using Thermus aquaticus 4-$\alpha$-glucanotransferase ($TA{\alpha}GTase$). Upon the enzyme treatment, the chain-length distributions of isoamylolytically debranched products became broader [degree of polymerization (DP): 3-40] than those of untreated corn starches. In addition, a variety of cycloamyloses (CAs) with different sizes were formed by the glucanotransfer activity of $TA{\alpha}GTase$. CAs with DP 5-40 were detectable in all of the $TA{\alpha}GTase$-treated corn starches. From the results of high-performance anion-exchange chromatography and high-performance size-exclusion chromatography analyses, it was suggested that the amount of CAs produced by the enzyme treatment increased as the amylose content of the starches increased. Thus, we concluded that the extent of modification of starch molecules was enhanced in proportion to amylose content by the transfer activity of $TA{\alpha}GTase$. This finding could be useful for developing an efficient process of CA production using this enzyme.

• 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.

• KSBB Journal
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• v.16 no.2
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• pp.128-132
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• 2001

Cyclodextrin glucanotransferase (EC 2.4.1.19 : 1,4-$alpha$-glucan 4-$alpha$-D-(1,4-glucano) transferase, cyclizing; CGTase) was recovered by starch adsorption. The adsorption and desorption of CGTase to starch was studied as a function of pH, temperature, and starch type. The optimal pH, temperature, and starch for adsorption were, 8.0, $4^{circ}C$, and 1% (w/v) corn starch, respectively, per 205 U/mL enzyme activity in the presence of 25% (w/v) ammonium sulfate. The maximum adsorption ratio was 95%. On the other hand, the optimal pH, temperature, and starch type for desorption were 8.0 (tris-buffer), $50^{circ}C$, and oxidized starch, respectively. The maximum desorption ratio was 98% by tris-buffer solution at pH 8.0. The efficiency of adsorption and desorption were affected slightly by the removal of cells from the fermentation broth.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.370-378
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• 1989

An Alkalophilic Bacillus circulans that can produce significant amount of cyclodextrin glucanotransferase (CGTase) was newly isolated from soil. The culture filtrate was successively purified by ($NH_4$)$_2$$SO_4$precipitation, DEAE-Sephadex column chromatography, and Sephadex G-100 column chromatography. The enzymatic properties, including molecular weight, optimal pH and temperature, stability, and kinetic parameters, were determined. The cyclodextrin synthesis reaction catalized by the purified CGTase was also studied. The sweet potato and corn starch were found to be the most suitable substrates with 60% conversion to cyclodextrin. The highest conversion was achieved at the CGTase concentration of 900-1,100 units/g of soluble starch. The purified CGTase could also catalize the transglycosylation on stevioside.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.78-81
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• 1998

Cyclodextrin glucanotransferase [CGTase, E.C.2.4.1.19] is an extracellular enzyme, which catalyzes he formation of ${\alpha}$-, ${\beta}$-, ${\gamma}$- CDs from starch. Their proportions of formations depend on enzyme sources and reaction conditions. To understand what determines the product specificity of CGTases, we examined the alteration of product specificity of CGTase from Bacillus macerans by organic solvent sand pH. At acidic pH range less than pH 6 where the enzyme was unstable, the ratio of ${\alpha}$-/ ${\beta}$-CD production was increased 4 times more than that at neutral pH range. As we increased the concentration of 2-butanol, ${\alpha}$-/ ${\beta}$-CD ratio was proportionally increased but / ratio remained constant. The ${\alpha}$-/ ${\beta}$-CD ratio of products was increased in the reaction media which yielded low products.

• Microbiology and Biotechnology Letters
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• v.26 no.5
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• pp.442-449
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• 1998

Several transglucosylated xylitols were synthesized using intermolecular transglucosylation reaction of cyclodextrin glucanotransferase (CGTase) and their bifidogenic effects were investigated. The CGTase from Thermoanaerobacter sp. showed the highest transglycosylation activity on xylitol compared to those obtained from other strains. Extruded starch was identified to be the most suitable glucosyl donor for transglucosylation reaction on xylitol molecule by CGTase. The optimum reaction conditions for transglucosylation were also studied using extruded starch as a glucosyl donor. The transglucosylated xylitols were purified by activated carbon column chromatography with ethanol gradient elution from 0 to 18%, and their chemical structures were analyzed by fast atom bombardment mass spectrometer, $\^$13/C-nuclear magnetic resonance spectrometer, and enzyme digestion method. Two transglucosylated xylitol, F-I and F-II, which had one or two glucose molecules attached to maternal xylitol by ${\alpha}$-1,4-linkage, were mainly obtained. F-II showed increased stimulation effect on the growth of Bifidobacterium breve compared to xylitol, indicating the possibility utilized as a new functional alternative sweetners having bifidogenic effects.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.368-376
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• 2003

Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of CGTase immobilization onto CNBr-activated sepharose 4B was as high as 18,000 units/g resin when the conditions was as follows: contact time 9 hrs at $37^{\circ}C$, pH 6.0, 100 nm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-$\alpha$-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature $37^{\circ}C$, 20% substrate solution containing 8% (w/v) of soluble starch and 12% (w/v) of L-ascorbic acid sodium salt, 100 rpm, far 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. Moreover the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.252-258
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• 1994

Transglycosylation of stevioside in the attrition coupled heterogeneous reaction system using raw starch as a glycosyl donor has significant advantages over conventional reaction systems using liquefied starch as a donor. The transglycosylation of stevioside under the presence of organic solvent showed that transglycosylation reaction occurs via two steps ; initially from raw starch to cyclodextrin(CD), and then followed by transglycosylation of produced CD. Comparison of the transglycosylation efficiency of c$\alpha $-, $\beta $, $\gamma $-CDs indicated that $\alpha $-, $\beta $-CD are mainly utilized as a glycosyl donor for following reaction. The reaction mechanism of transglycosylation between stevioside and CD proceeded according to random sequential bireactant mechanism. The equilibrium constant of transglycosylation reaction of cyclodextrin glucanotransferase wase also evaluated. The structure of transglycosylated stevioside was confirmed by TLC, and it was found that glycosyl group(G$_{1}, $ ~ G$_{4}$-glycosidic bond.

• 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.

• 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.