• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.972-978
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• 2011

In this study, we investigated the performance of an immobilized ${\beta}$-galactosidase inclusion bodies-containing Escherichia coli cell reactor, where the cells were immobilized in alginate beads, which were then used in repeated-batch operations for the hydrolysis of o-nitrophenyl-${\beta}$-D-galactoside or lactose over the long-term. In particular, in the Tris buffer system, disintegration of the alginate beads was not observed during the operation, which was observed for the phosphate buffer system. The o-nitrophenyl-${\beta}$-D-galactoside hydrolysis was operated successfully up to about 80 h, and the runs were successfully repeated at least eight times. In addition, hydrolysis of lactose was successfully carried out up to 240 h. Using Western blotting analyses, it was verified that the ${\beta}$-galactosidase inclusion bodies were sustained in the alginate beads during the repeated-batch operations. Consequently, we experimentally verified that ${\beta}$-galactosidase inclusion bodies-containing Escherichia coli cells could be used in a repeated-batch reactor as a biocatalyst for the hydrolysis of o-nitrophenyl-${\beta}$-D-galactoside or lactose. It is probable that this approach can be applied to enzymatic synthesis reactions for other biotechnology applications, particularly reactions that require long-term and stable operation.

• KSBB Journal
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• v.9 no.4
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• pp.400-405
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• 1994

The conditions for ${\beta}$-galactosidase production from alkalophilic, thermophilic Bacillus sp. TA-11 were investigated. The maximal enzyme production was obtained when the strain was cultured at $50^{\circ}C$ for 5 days with fed-batch culture in the optimal medium containing 1.5% lactose, 0.6% yeast extract 0.15% $K_2HP0_4$and initial pH 9.5, and then final enzyme activity under the above conditions was 5200 unit/ml of cell free extract.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.3
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• pp.465-468
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• 2002

The galactose/mannose ratio of guar gum, guar gum treated with purified ${\alpha}$-galactosidase and locust bean gum were investigated. Gel-promoting property of enzyme-treated guar gum increased when the galactose/mannose ratio was about 1 : 3.2, which was close to the ratio of 1 : 3.3 for locust bean gum. And the ratio was obtained when the guar gum was hydrolyzed by the enzyme for 24 hr. It is clear that enzymatic depletion of galactose from guar gum by sunflower seed ${\alpha}$-galactosidase would lead to a significant increase in gelation ability. The mixture of xanthan gum and guar gum, and xanthan gum, guar gum and enzyme-treated copra meal were also investigated in viscosity behavior.

• Food Science and Preservation
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• v.2 no.1
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• pp.173-183
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• 1995

This paper was carried out to investigate changes in chromatograms of polysacctatides and soluble pectins on Sephadex G-50 and non-cellulosic neutral sugars of polysaccharides isolated from cell wall of persimmon fruits treated with polygalacturonase and $\beta$-galactosidase in vitro. The chromatogram pattern of soluble pectins extracted from cell wall treated with $\beta$-galactosidase on Sephacryl S-500 column were similar to those of untreatment, but contents of soluble pectins treated with $\beta$-galactosidase were different from those of untreatment. The patterns of chromatograms In soluble pectins extracted from cell wall treated with polygalacturonase were more complex and lower molecular polymer than those of other cell wall-degrading enzyme treatments. Non-cellulosic neutral sugar of polysaccharides in fraction I of soluble material treated with polygalacturonase was rhamnose, those in fraction II were similar to those in fraction III and contents of arabinose, xylose and glucose were higher than contents of other non-cellulosic neutral sugars. Non-cellulosic neutral sugars of polysaccharides in fraction I in soluble material by $\beta$-galactosidase treatment were rhamnose, arabinose, galactose and mannose. Content of glucose of polysaccharides in fraction II was higher than that in fraction I . Non-cellulosic neutral sugars treated with mixed enzyme were rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose. Compositions of non-cellulosic neutral sugars of polysaccharides in fraction I were similar to those in fraction II and III.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.3
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• pp.373-375
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• 1993

Quantitative ${\beta}$-galactosidase estimation in the intestinal mucosal cells of calves with diarrhea under experimental conditions due to rotavirus were undertaken. A quantitative decrease of 40-70% in ${\beta}$-galactosidase activity was observed in proximal and middle segments of the small intestine of the infected calves, more so in the middle segments. The decrease in the distal part of the intestine, however, was lesser (5 to 30%). The decrease in the activity was more marked on the day 2 to 6 post infection indicating the degree of the damage of the villi of the small intestine.

• Journal of Microbiology and Biotechnology
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• v.4 no.3
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• pp.200-203
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• 1994

Spodoptera frugiperda IPLB-SF-21-AE cells were cultivated in a spin-filter bioreactor with continuous perfusion for the recombinant $\beta$-galactosidase production. At the perfusion rate of 0.06 $hr^{-1}$, the maximum cell density of insect cells in this bioreactor system reached 3.5$\times$$l0^6$ viable cells/ml using the Grace media containing 5% FBS and 0.3% Pluronic F-68. The recombinant $\beta$-galactosidase production of 8, 100 units per reactor volume was also achieved at this perfusion rate.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.509.1-509
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• 1986

Recent advances in recombinant DNA techniques have provided a tool for breeding of microorganisms of hyper production. Enzyme production by cloned microorganism has some advantages. They are ⅰ) Enzymes can be produced by a microorganism easily cultured ⅱ) Hyper production. ⅲ) In some cases, such as thermophilic enzyme gene is cloned in a mesophilic bacteria, the enzyme purification procedure can be simplified. One example, production of thermophilic ${\beta}$-galactosidase in B. subtilis will be presented. Bacillus stearothermophilus IAM 11001 produced three ${\beta}$-galactosidases, ${\beta}$-galactosidase I, II and III (${\beta}$-gal-I, II and III). By connecting restriction fragments of the chromosomal DNA to plasmid vector, followed by transformation of Escherichia coli, two ${\beta}$-galactosidase genes (bgaA and bgaB) located close to each other on the chromosome were cloned.

• KSBB Journal
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• v.15 no.1
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• pp.84-87
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• 2000

We studied the effect of ultrasoft X-ray obtained from the Pohang Light Source (PLS), on the mutation of E. coli and the damage of plasmid. After irradiation, the supercoiled plasmid DNA converted to the relaxed-form, and then to the linear-form. We transformed the irradiated plasmid DNA and isolated $\beta$-galactosidase mutants. We also isolated $\beta$-galactosidase mutants from the directly irradiated cells. There were preferred mutational sites on DNA induced by ultrasoft X-ray.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.10a
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• pp.384-387
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• 2004

겨울철 토양에서 ${\beta}-Galactosidase$를 생산하는 균주를 분리하였으며 동정한 결과 그람 음성 간균이고 Pantoea spp. 로 확인되었다. Pantoea spp. 균주의 세포 추출물로부터 DEAE-Sephacel chromatography와 affinity chromatography를 이용하여 ${\beta}-Galactosidase$를 분리하였다. Pantoea spp. 의 ${\beta}-Galactosidase$의 반응 최적 온도는 $45^{\circ}C$이이고 최적 pH는 $5.5{\sim}7.5$이고 열안정성을 조사한 결과 $45^{\circ}C$이상의 온도에서 불활성 되는 것으로 나타났고 E. coli에서 분리된 효소보다 저온에서의 활력이 좋았지만 상업적인 효소인 Kluyveromyces lactis (Validase) 보다는 낮았다.

• Journal of Microbiology and Biotechnology
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• v.14 no.4
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• pp.863-867
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• 2004

$\alpha$-Galactosidase from A. oryzae DR-5 was induced in the presence of melibiose, raffinose, galactose, and locust bean galactomannan. The enzyme was purified to homogeneity by precipitation with acetone followed by ion-exchange chromatography using DEAE-Sephacel. The purified enzyme showed a single band in both nondenaturing-PAGE and SDS-PAGE. The enzyme was a glycoprotein in nature by activity staining. The molecular weight of the purified enzyme was 93-95 kDa by SDS-PAGE. The enzyme exhibited the optimum pH and temperature at 4.7 and $60^\circ{C}$, respectively. $\alpha$-Galactosidase activity was strongly inhibited by $Ag^{2+}, Hg^{2+}, Cu^{2+}$, and galactose. EDTA, 1,10-phenanthraline, and PMSF did not inhibit the enzyme activity, whereas N-bromosuccinimide completely inhibited enzyme activity. Investigation by TLC showed complete hydrolysis of stachyose and raffinose in soymilk in 3 h at pH 5.0 and $50^\circ{C}$.