• Food Industry And Nutrition
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• v.2 no.1
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• pp.31-41
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• 1997

Stevioside 는 남미 파라과이 원산의 국화와 식물인 Ste-via rebaudiana Bertoni 유래의 천연대체감미료서 식품, 의약품, 주류 산업에 널리 활용되고 있다. 이와 같은 stevio-side 의 구조와 특성, 제조방법, 그리고 사용현황 등을 기술하였으며, 당전이 stevioside의 종류 제조방법을 정리하였다. 본 연구실에서 개발한 생전분을 당공여체로 히용하는 분쇄마찰매체 효소반응계와 팽윤 extrusion 전분을 당공여체로 이용하는 불균일상 효소반응계에서의 stevioside 당전이반응의 특성과 산업적 활용 가능성을 검토하였다. 상기 두 종류의 불균일상 효소반응계는 높은 수율, 반응속도, 낮은 maltooligo 당의 축적, 용이한 당전이 stevioside 의 분리정제 등 기존의 액화 전분을 당공여체로 이용하는 반응계에 비하여 많은 장점이 예상되는 산업적 활용 가능성이 높고 고효율 반응계임을 알 수 있었다. 끝으로 stevioside 감미료의 장래 전망에 관하여 예측하였다.

• KSBB Journal
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• v.9 no.2
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• pp.108-114
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• 1994

Kinetic equations for transglycosylation of stevioside in the attrition coupled reaction system using raw starch as a glycosyl donor were derived considering that the reaction was carried out through two steps; production of cyclodextrin(CD) from raw starch in the attrition coupled reaction system and then transglycosylation of glycosyl residues to stevioside from produced CD. Kinetic constants of derived equation were evaluated. The simulation result showed that the derived kinetic equations could predict the experimental data reasonably well and that can be utilized for optimization and scale-up of transglycosylation reactor and process developments.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.802-806
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• 2000

The transglycosylation reaction by dextansucrase from Leuconostoc mesenteroides B512FMC/6HG8 was investigate with cellobiose as an acceptor molecule and sucrose as a donor. he optimal conditions of transglycosylation on cellobiose were found that the ration of sucrose to cellobiose was 3:1, the amount of enzyme was 2U/mL, the ionic strength of buffer was 25 mM, pH was 5.0 and reaction temperature was $25^{\circ}C$. also, acceptor products of cellobiose by transglycosylation were a series of oligosaccharides showing the degree of plymenzation of 6.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.249-254
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• 1997

Stevioside was transglucosylated to improve its sweetness. Eighteen conditions were tested using the tables of orthogonal arrays of $L_{18}\;(2^{1}{\times}3^{7})$. Statistical analysis showed that the transglucosylation rate was significantly affected by temperature, the ratio of co-substrate (maltodextrin) to stevioside, pH, DE of maltodextrin and concentration of stevioside, in their order. Optimum conditions selected for temperature, ratio of co-substrate to stevioside, pH and DE of maltodextrin were $80^{\circ}C$, 1.0, 6.0 and 15, respectively. Glycosyl-stevioside with 68% yield of transglucosylation was produced at the optimum condition and found to have better quality in sweetness than stevioside and rebaudioside A.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.10
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• pp.1417-1422
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• 2012

Alkyl glucosides were synthesized using the transglycosylation reaction of Celluclast, the cellulase from Trichoderma reesei, with cellobiose and various alcohols. Glucose as a by-product of the reaction was removed using the immobilized yeast system. Among the alkyl glucoside products, the acceptor products of methanol and ethanol were confirmed as methyl ${\beta}$-D-glucopyranoside and ethyl ${\beta}$-D-glucopyranoside via MALDI-TOF MS and enzymatic analysis. Optimal yields of methyl ${\beta}$-glucoside and ethyl ${\beta}$-glucoside were 65.3% (mol/mol) and 59.0% (mol/mol), respectively, based on cellobiose consumed.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.1-6
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• 1999

Benzylalcohol-$\alpha$-glucoside (BG) was synthesized from soluble starch by transglycosylation of $\alpha$-amylase. Transglycosylation in water-organic two phase system containing 1% soluble starch as a glycosyl donor, 90% benzylalcohol as a glycosyl acceplor, 10% citrate buffer solulion (0.1 M, pH 5.0), and 10 unit of $\alpha$-amylase (Aspergilllw oryzae) was showed highcst efficiency. About 4 mg BG was obtained from 10 mg starch in reaction for 80 hrs at $40^{\circ}C$. Initially benzylalcohol-$\alpha$-maltoside Q3M) was major product, but as the reaction proceeded, it was hydrolyzed to glucose and BG. Finally the product of transglycosylation by $\alpha$-amylase was only BG. The both products did not show reducing powcr and hydrolyzed by $\alpha$-glucosidase and $\alpha$-amylase, respectively. The molecular wcights of both were estimated to be 270 and 432 by ES1-Mass, respectively.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.29-36
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• 2004

$\alpha$-Cyclodextrin glucanotransferase excreted from a newly isolated Geobacillus thermosacchalytycus was purified through the ultrafiltraion, hydrophobic Sepharose CD-4B affinity chromatography, and gel filtration on Sephadex G-75, respectively. The molecular weight of the purified CGTase was 69 kDa and its N-terminal amino acid sequence was determined to be Asn-Leu-Asn-Lys-Val-Asn-Phe-Val-Ser-Asp-Val-Val-Val-Gln-Ile. The optimum pH and temperature were pH 6.0 and$ 60^{\circ}C$, respectively, and stably at the pH range of 6.0-8.0 and $60^{\circ}C$ in the presence of $Ca^{++}$. The excreted CGTase from the thermophilic G. thermosacchalytycus was $\alpha$-type showing a high coupling activity for the transglycosylation on various glucosides. The coupling reaction was carried out according to the random ternary complex mechanism.m.

• 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.26 no.2
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• pp.187-194
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• 1998

Glycosides were synthesized using transglycosylation reaction of amylase in water system. Starch as a glycosyl donor and benzylalcohol as an acceptor were selected as substrates of transglycosylation reaction. Among tested 9 commercial amylase, amyloglucosidase from Rhizopus sp. had high activity for transglycosylation from starch. The glycoside synthesized in water phase by amyloglucosidase was identified as benzylalcohol-${alpha}$-glucoside (BG) of which one molecule of benzylalcohol was bound to 1-OH of glucose. The transglycosylation reaction by amyloglucosidase were carried out in reaction system containing 50 mg starch, 50 mg benzylalcohol, and 10 units enzyme in pH 5.0 at 45$^{\circ}C$. The synthesized BG was hydrolyzed by ${alpha}$-glucosidase to produce glucose and benzylalcohol.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.104-110
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• 2012

Glycosynthase is an active site nucleophile mutant enzyme, prepared from glycosidase, which is capable of synthesizing oligosaccharide derivatives without the hydrolysis of the product. Thermoacidophilic ${\alpha}$-glucosidase of Thermoplasma acidophilum (AglA) exhibits a transglycosylating activity yielding various glycosides. AglA was converted to glycosynthase by the substitution of the catalytic nucleophile Asp-408 residue into non-nucleophile glycine in order to increase its ability to synthesize various glycosides by transglycosylation. The glycosynthase mutant was purified by Ni-NTA chromatography and its glycoside-synthesizing activity was measured by using an external nucleophile, sodium formate buffer, providing maltose as a donor and p-nitrophenyl-${\alpha}$-D-glucopyranoside ($pNP{\alpha}G$) as an acceptor, respectively. In addition, $pNP{\alpha}G$ was examined for its feasibility to act as both a donor and an acceptor, and products were compared with those of the wildtype enzyme. The mutant enzyme was found to catalyze the formation of a specific product from $pNP{\alpha}G$ with a yield of 42.5% without further hydrolysis, while the wild-type enzyme produced two $pNP{\alpha}G$ products at low yields. The results demonstrate the possibility of satisfactory yields for the reactions in the presence of small amounts of acceptor, and demonstrate that the high activity of the mutant, at pHs below neutrality, was applicable in the transfer of glucose from the natural donor.