• Journal of Microbiology and Biotechnology
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• v.7 no.3
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• pp.204-208
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• 1997

Aureobasidium puliulans possesses the capacity for simultaneous formation of fructosyltransferase and glucosyltransferase in various sugar media including sucrose, maltose, glucose and fructose. Among them, sucrose (300 g/1) was the most suitable carbon source for fructosyltransferase production, while fructose (100 g/1) gave the maximal production of glucosyltransferase. There existed a critical concentration for the optimal formation of enzymes in sucrose, glucose and fructose media. By contrast, no effect of maltose concentrations up to 300 g/1 was observed. The specific activity of the glucosyltransferase on maltose medium was highest during the early period of fetmentation, after which a sharp decrease occurred, whereas fructosyltransferase activity on sucrose medium maintained a nearly constant rate for a given culture period. Concomitant production of fructosyltransferase and glucosyltransferase was investigated with different combinations of lower concentrations of sucrose and maltose. Maltose supplementation in sucrose media and sucrose addition to maltose media enhanced the activity ratios of fructosyltransferase to glucosyltransferase as compared to that of non-supplemented media. Several polymers and surfactants were added in an attempt to enhance enzyme production, and supplementation of polyoxyethylene-sorbitan monolaurate (Tween 20) promoted fructosyltransferase production by 20%.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.3
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• pp.534-539
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• 1997

In order to reuse enzyme efficiently, a mthod for ionic binding of fructosyltransferase to a porous carrier bearing quaternary alkyl alkanolammonium groups was investigated. The fructosyltransferase activity of the immobilized enzyme increased with increasing amount of loaded enzyme, and maximally reached 770U/g of the carrier when loaded amount of the enzyme was 18.2 mg/g carrier. The immobilized fructosyltransferase had optimum pH and temperature of 7.5 and 45$^{\circ}C$, respectively, whereas soluble enzyme had 6.5 and 55$^{\circ}C$: the Km value for the immobilized enzyme was 27.8 mM for sucrose, which was the same as that of soluble enzyme. In a batch reactor, the enzyme produced a mixture of fructooligosaccharides, mainly F$_2$G, from sucrose with the slight loss of enzyme activity during continuous operation of 12 days at 42$^{\circ}C$.

• KSBB Journal
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• v.9 no.1
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• pp.40-47
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• 1994

A simplified mathematical model for the production of high-purity fructo-oligosaccharides by the mixed-enzyme system of fructosyl transferees and glucose oxidase was proposed and compared with the experimental results. The kinetic parameters including $K_m,\;V_{max}\;and\;K_{iG}$ were estimated at $40^{\circ}C$, in which $K_m$, values decreased and $K_{iG}$ and $V_{max}$ values increased compared with those of fructosyl transferees alone. The kinetics of the mixed-enzyme system was successfully described in the form of Michaelis-Menten equations. At the reasonable sucrose concentrations tested, the simulated sugar profiles were of good agreement with the experimental ones.

• KSBB Journal
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• v.8 no.4
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• pp.307-312
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• 1993

A fructosyltransferase from Aureobasidium pullulans was immobilized onto a polystyrene-type anionic ion-exchange resin and the production of fructo-oligosaccharides was Investigated by the immobilized enzyme. The optimum pH and the temperature of immobilized enzyme were found to be pH 5.0, $55^{\circ}C$ respectively. The thermal stability of the enzyme was greatly enhanced after immobilization. The reaction profiles of the immobilized enzyme was almost identical to those of the free cells and the soluble enzyme. The immobilized enzymes were stable up to 20 cycles without loss of initial activity in a repeated-batch operation $50^{\circ}C$.

• KSBB Journal
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• v.10 no.1
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• pp.9-14
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• 1995

Continuous production of fructooligosaccharides from sucrose by a dual immobilized enzyme system of fructosyltransferase and glucose isomerase was studied in a column reactor. The optimal temperature and pH of the immobilized fructosyltransferase were $65^{\circ}C$ and 5.5, respectively. The activity of glucose isomerase was favorable as temperature and pH were increased within the ranges examined. However, both the immobilized enzymes were thermally unstable over $5^{\circ}C$, suggesting that long-term operation of the dual immobilized enzyme column should be carried out below $50^{\circ}C$. The optimum packing ratio of fructosyltransferase to glucose isomerase was found to be around 5/3. Under the optimized reaction conditions, the dual enzyme column was successfully operated for 40 days without any loss of initial enzyme activities, yielding 66% of fructooligosaccharides. Furthermore, the relative sweetness of fructooligosaccharides produced by a dual emzyme system was enhanced by 6% compared with that of fructosyltransferase alone.

• Biotechnology and Bioprocess Engineering:BBE
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• v.1 no.1
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• pp.18-21
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• 1996

A continuous production of fructooligosaccharides from sucrose was investigated by fructosyltransferase immobilized on a high porous resin, Diaion HPA25. The optimum pH(5.5) and temperature(55$^{\circ}C$) of the enzyme for activity was unaltered by immobilization, and the immobilized enzyme became less sensitive to the pH change. The optimal operation conditions of the immobilized enzyme column for maximizing the productivity were as follows: 600g/L of sucrose feed concentration, flow rate of superficial space velocity 2.7h-1. When the enzyme column was run at 50$^{\circ}C$, about 8% loss of the initial activity of immobilized enzyme was observed after 30 days of continuous operation, during which high productivity of 1174g/L$.$h was achieved. The kinds of products obtained using the immobilized enzyme were almost the same as those using soluble enzymes or free cells.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.527-529
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• 2003

A fructosyltransferase from Leuconostoc mesenteroides TL1 was purified. The molecular mass of the enzyme was estimated to be 38 kDa based on the activity staining. The pH and temperature optima of the enzyme were 6.3 and $40^{\circ}C$, respectivly. Structural analysis of the polymer prepared from sucrose by the enzyme was determined by NMR Spectroscopy: It shows the heterogeneous linkages of levan-like fructan and dextran-like glucan.

• KSBB Journal
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• v.19 no.5
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• pp.368-373
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• 2004

The optimal condition for the production of a glucan and a fructan synthesizing enzymes from Leuconostoc mesenteroides NRRL B-1149 were studied based on the different medium compositions. Response surface methodology was applied to find the optimistic condition showing the relationship between the fermentation response (enzyme activities) and the fermentation variable concentrations of yeast extract, peptone concentration, K2HP04 concentration and sucrose. Optimum medium composition for both enzymes production was $0.75\%$ yeast extract, $0.72\%$ peptone, $1\%$ K2HP04 and $2.17\%$ sucrose. Using this medium, the activities produced in culture was 0.90 U/m~ for glucosyltransferase (GTase) and 0.96 U/ml for fructosyltransferase (FTase). After purification of 1149FTase by consecutive chromatographies using Sephadex G-150 and DEAE-Sepharose, a 1149FTase of 210 kDa on $7\%$ polyacrylamide gel was isolated and it synthesized soluble fructan. The 1149GTase showed a band of 180 kDa on $8\%$ polyacrylamide gel after purification using Bio-Gel P-100 gel chromatography and DEAE-Sepharose ion exchange chromatography and it synthesized insoluble glucan. The linkages of polymers were determined by methylation using Hakomori reagent and following NMR analysis. The glucan was composed of a(1~6) and a(1~3) linkages and the fructan was levan.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.100-104
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• 2006

In this study, we investigated the relationship between the morphology and the rheological properties of Penicillium citrinum to improve the production of neo-fructosyltransferase (neo-FTase). In a 2.5 L bioreactor culture of P. citrinum, it was observed that agitation speed and aeration rate had significant effects on the production of neo-FTase and that maximum cell mass and neo-FTase production obtained at 500 rpm and 1.5vvm were 8.14 g/L and $53.2{\times}10^{-3} U/mL$, respectively. Cell mass and neo-FTase production increased to 91.53 and 25.17%, respectively. In the morphology and rheology studies, P. citrinum showed a typical pellet morphology that was explained by a shaving mechanism; this phenomenon was significantly affected by carbon sources. The rheology of neo-FTase fermentation by P. citrinum was dependent on cell growth and fungal morphology.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.188-191
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• 2002

A glucan and a fructan producing enzymes from Leuconostoc mesenteroides NRRL B-1149 were prepared and concentrated from the cu1ture of 1.5% sucrose using polysulfone ultrafiltration hollow fiber in the presence of 0.1% (w/v) Tween 80, 1 mM $CaCl_2$, and 0.02% $NaN_3$. The molecular masses of the enzymes were estimated to be about 213.6 kDa and 180 kDa, respectively, based on the PAS staining for the glucosyltransferase and Mukasa method for fructosyltransferase. Polymers produced by the enzymes showed different solubility; an insoluble glucan and a soluble fructan. The linkages of polymers were determined by methylation using Hakomori reagent and following acid hydrolysis. The glucan was composed of ${\alpha}$-1,6 and 1,3 linkages and the fructan showed similar linkage data of levan.