• 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.

• Korean Journal of Food Science and Technology
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• v.26 no.4
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• pp.375-381
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• 1994

A bacterial strain No. KY-126, which produced extracellular cyclodextrin glycosyltransferase(CGTase), was isolated from soil and identified as Bacillus stearothermophilus KY-126. The enzyme was purified by the treatments of ammonium sulfate precipitation, DEAF-Sephadex, Sephadex G-100 column chromatography. The optimal pH and temperature for the enzyme activity were pH 5.5 and $65^{\circ}C$, respectively. And the enzyme was stable at pH values from 6.0 to 11.0 at $55^{\circ}C$ for 30 min and stable up to $60^{\circ}C$ for 30 min.. The enzyme was inhibited by $HgCl_{2}$. The molecular weight of the enzyme was estimated to be 67,000 by using SDS-PAGE. The maximum conversion from starch to cyclodextrin (CD) by CGTase was 43% and obtained at 6 hr reaction and the ratio of ${\alpha}-,\;{\beta}-,\;{\gamma}-$, CD production at this time was 2.9 : 2.1 : 1.0.

• Proceedings of the Membrane Society of Korea Conference
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• 1998.10a
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• pp.82-85
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• 1998

1. 서론 : Cyclodextrin(CD) 동족체(homologues)는 $\alpha$-, $\beta$-, $\gamma$-CD로 구분되며, 이들 각각은 $\alpha$-D-glucopyranose 단위체 6,7, 및 8개가 비환원성 환상구조로 연결된 cyclic maltooligosaccaride의 일종으로 외부는 친수성이고, 내부는 소수성인 공동 구조를 갖고 있다. 따라서 각 CD는 동공의 크기가 달라 다른 크기의 소수성 물질들과 선택적인 포접화합물 (inclusion compound)을 형성하는 특징이 있다. CD 동족체는 전분 분해 효소인 cyclodextrin glycosyltransferase(CGTase)에 의해 전분으로부터 생산되는데, 반응용액 내에서의 CD 동족체 농도가 어느 한계값 이상으로 높아지면 생산물 저해와 다른 환원당으로의 분해 때문에 생산성이 감소하여 이의 효과적 생산에 어려움이 있다. 본 연구는 dead-end 및 cross-flow형 막 생물반응기를 사용하여 CGTase에 의한 전분의 CD 동족체로의 분해반응시 생산물 저해를 억제시켜 생산성을 향상시키고, 동시에 조작조건 변화에 따른 생산물인 CD 동족체의 효과적인 연속분리 가능성을 검토하였다.

• Membrane Journal
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• v.8 no.3
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• pp.170-176
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• 1998

A study on the bioconversion of soluble starch to cyclodextrin(CD) homologue by CGTase was performed in the membrane-enzyme reactor equipped with a dead-end type membrane module. in the batch reactor, the total conversion of soluble starch to CD homologue was decreased rapidly from a maximum value of 45 % with increasing reaction time due to the product inhibition and breakdown of CD homologue to the reducing sugars. However, in the membrane-enzyme reactor, the total conversion of soluble starch was maintained at a constant value of 35 % throughout the reaction, since the membrane(MWCO = 10,000) promptly separated CD homologue from the reaction mixture. After the macdon for 24 hr in the membrane-enzyme reactor using a 10 % soluble starch solution, the cumulative production amount of CD homologue was about 3.7 kg/m$^2$ at the operating pressure of 2 atm.

• Korean Journal of Food Science and Technology
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• v.23 no.4
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• pp.503-509
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• 1991

Action of a cyclodextrin glycosyltransferase (CGTase) produced from alkalophilic Bacillus sp. No.4 was studied in a solution containing starch and sucrose to prepare glycosyl sucrose syrup with good sweetness and antidecaying properties of teeth. In the initial stage of the reaction the CGTase produced cyclodextrin, however, the cyclodextrin disappeared and glycosyl sucrose was formed with the lapse of reaction time. The best proportion of sucrose to starch for prodution of glycosyl sucrose was about 1 : 1. The optimum pH and temperature of the coupling reaction was pH 6.0 and $60^{\circ}C$, respectively. Main composition of glycosyl sucrose syrup prepared with 20% starch and 20% sucrose was sucrose 18%, glucosyl sucrose ($G_{2}F$) 15.3% and maltosyl sucorse ($G_{3}F$) 11.3%. And glucose, maltose and maltotriose were produced very little. Smaller amounts of acid and insoluble glucan were formed in the syrup by Streptococcus mtans OMZ176 than in the sucrose. Therefore, the prepared glycosyl sucrose sucrose syrup is expected to prevent teeth from decaying.

• 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 ${\gamma}-cyclodextrin$.

• The Korean Journal of Food And Nutrition
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• v.8 no.3
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• pp.253-261
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• 1995

A strain of alkalophilic Bacillus sp. C-21 has been Isolated from sold. The strain was capable of producing large amount of cyclodextrin glycosyltransferase (CGTase) in the high alkaline pH medium. The preferable medium composition was determined to be as follows : 1.0% soluble starch, 1.0% peptone, 0.5% yeast extract, 0.1% K2HP04, 0.02% MgSO4.7H2O and 1.0% Na2CO3(pH 10.0) The highest enzyme production was observed after 30hours of cultivation at 33$^{\circ}C$. The optimum temperature and pH for the activity of crude enzyme were 6$0^{\circ}C$ and 6.0, respectively. The enzyme was stable between pH 6.0 and 9.6, and up to 55$^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.148-153
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• 1989

A strain of alkalophilic Bacillus sp. YC-335 has been isolated from soil. The strain was capable of producing large amount of cyclodextrin glycosyl transferase (CGTase) in the culture broth. The preferable medium composition has been determined to be as follows : 1.5% soluble starch, 5% corn steep liquor, 0.1% $K_2$HPO$_4$, 0.02%mgSO$_4$.7$H_2O$, 1% CaCO$_3$and 1% Na$_2$CO$_3$(pH 10.3). The highest enzyme production was observed after 48 hours of cultivation at 31$^{\circ}C$. The optimum pH and temperature for the activity of crude enzyme were 6.0 and 5$0^{\circ}C$, respectively. The enzyme was stable between pH 5 and 9, and upto 5$0^{\circ}C$. The enzyme converted starch into $\alpha$-, $\beta$- and ${\gamma}$-CD in the relative amounts of 1:10:1.5, respectively.

• Microbiology and Biotechnology Letters
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• v.23 no.4
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• pp.425-431
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• 1995

A kinetic model of the cyclodextrin formation in a heterogeneous enzyme reaction system using swollen extrusion starch as substrate was derived emphasing the structural features of extrusion starch. The degree of gelatinization, the ratio of accessible and inaccessible portion of extrusion starch, adsorption of CGTase on swollen starch, the structural transformation during reaction, and product inhibition caused by produced CDs were considered in deriving kinetic model. Various kinetic constants were also evaluated. The derived kinetic equation was numerically simulated, which result showed that the derived kinetic equations can be used to predict the experimental data reasonably well under the various experimental conditions. Kinetic model can be utilized for the optimization of enzyme reactor and the process development for CD production from swollen extrusion starch.

• 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.