• Korean Journal of Food Science and Technology
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• v.25 no.6
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• pp.689-693
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• 1993

${\alpha}-galactosidase$ releases galactoside from raffinose and stachyose which are the major sugars in soybean, Although raffinose and stachyose were known as flatulence factors, these sugars were recently claimed as bifidus factors. In this experiment we studied the properties of ${\alpha}-galactosidase$ and its production from Bifidobacterium sp. Int-57. Int-57 produced higher level of ${\alpha}-galactosidase$ than other intestinal bacteria. The production of ${\alpha}-galactosidase$ was greater when grown on raffinose compared with other carbohydrates tested. Partially purified ${\alpha}-galactosidase$ was obtained after sonication of harvested cell pellet followed by DEAE-cellulose chromatography and Sepharose CL-6B gel filtration, and assayed using PNP-${\alpha}-galactosidase$ as a substrate. Optimum pH for activity was 7.0 and optimum temperature was $40^{\circ}C$. At 5 mM concentration of metal ions, $CoCl_{2}\;and\;CuCl_{2}$ and inhibited the enzyme activity by 33% and 21% respectively. The enzyme was shown to hydrolyse genuine substrates, i.e. raffinose and stachyose.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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• pp.552-557
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• 2013

Partially purified ${\alpha}$-galactosidase from Bacillus sp. LX-1 was non-covalently immobilized on a reversibly soluble-insoluble polymer, Eudragit L-100, and an immobilization efficiency of 0.93 was obtained. The optimum pH of the free and immobilized enzyme was 6.5 to 7.0 and 7.0, respectively, while there was no change in optimum temperature between the free and immobilized ${\alpha}$-galactosidase. The immobilized ${\alpha}$-galactosidase was reutilized six times without significant loss in activity. The immobilized enzyme showed good storage stability at $37^{\circ}C$, retaining about 50% of its initial activity even after 18 d at this temperature, while the free enzyme was completely inactivated. The immobilization of ${\alpha}$-galactosidase from Bacillus sp. LX-1 on Eudragit L-100 may be a promising strategy for removal of ${\alpha}$-galacto-oligosaccharides such as raffinose and stachyose from soybean meal and other legume in feed industry.

• Journal of Applied Biological Chemistry
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• v.44 no.2
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• pp.53-58
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• 2001

Glycosidase activities in the grape flesh (Marguerite) were assayed, and the order of activity was marked as follows: ${\alpha}$-D-galactosidase>${\alpha}$-D-mannosidase>${\alpha}$-D-glucosidase>${\beta}$-D-galactosidase>${\beta}$-D-glucosidase. Of these glycosidases, ${\alpha}$- and ${\beta}$-D-galactosidases were prominently expressed by the treatment of gibberellic acid, resulting in 56 and 238% increase of activity, respectively. Most of ${\alpha}$-D-galactosidase was found in the flesh texture, and the activity increase by gibberellic acid occurred mostly in this tissue. The increase in ${\alpha}$-D-galactosidase activity was dependent on the concentration of gibberellic acid treated, showing a positive correlation. Gibberellic acid affected the content of total protein in the grape flesh, 49% increase by 75 ppm treatment. Above this concentration, higher gibberellic acid level did not influence the protein expression. Specific activity of the ${\alpha}$-D-galactosidase still increased, showing 24% increase in activity. Grape flesh subjected by gibberellic acid (100 ppm) resulted in the increased activity against a natural substrate, stachyose, showing 55% increase in activity from the grapes treated with 100 ppm of gibberellic acid. Other natural substrates, such as melibiose and raffinose, were also considerably hydrolyzed, and the extent was similar to that of stachyose hydrolysis. During postharvest storage, ${\alpha}$-D-galactosidase activity in the grape flesh increased by 51% after 20 days and then declined slowly.

• Journal of Ginseng Research
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• v.24 no.2
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• pp.89-93
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• 2000

WB were screening the stele and the cortex of the ginseng roots (Panax ginseng C.A.Meyer) on the exo-0-glycosylhydrolase activities during vegetation period of 1999 year. The following p-nitrophenylglycosides were used to test exe-0-glycosylhydrolase activities: $\alpha$- and $\beta$-D-galactopyranosides,$\alpha$- and $\beta$-D-glucopyranosides, $\alpha$- and $\beta$-D-mannopyranosides, N-acetyl-$\beta$-D-glucosaminide, $\alpha$- and $\beta$-D-xylopyranosides $\alpha$- L-rhamnopyranoside, $\beta$-D-glucuronide, $\beta$-D-galacturonide, $\beta$-L-,$\alpha$-L- and $\beta$-D-fucopyranosides, $\alpha$-L-arabinopyranoside. Only $\beta$-D-galactosidase, $\alpha$-L-mannosi-dase , N- acetyl- ${\beta}$-D-slucosarninidase, $\alpha$-D-galacto sidase, $\alpha$-L-arabinosidase, and $\beta$-D-fuco sidase were found in both partsof ginseng roots. Their contents during the vegetation period were shown to differ considerably, being dependent not only on plant development stage but on plant tissue and environmental conditions too.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.700-705
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• 2014

A 12-week feeding trial was conducted with 87 g rainbow trout to evaluate the effects on growth performances, feed efficiency and nutrient digestibility of adding ${\beta}$-mannanase and ${\alpha}$-galactosidase enzymes, solely or in combination. Seven diets were prepared by adding ${\beta}$-mannanase, ${\alpha}$-galactosidase and mixed enzyme at two different levels (1 g/kg and 2 g/kg) to control diet (without enzyme) including soybean meal. Mixed enzymes (1 g/kg, 2 g/kg) were prepared by adding ${\beta}$-mannanase and ${\alpha}$-galactosidase at the same doses (0.5+0.5 g/kg and 1+1 g/kg). At the end of the experiment, addition of ${\beta}$-mannanase, ${\alpha}$-galactosidase and mixed enzyme to diet containing 44% soybean meal had no significant effects on growth performance and gain:feed (p>0.05). In addition, adding ${\beta}$-mannanase, ${\alpha}$-galactosidase and mixed enzyme in different rations to trout diets had no affect on nutrient digestibility and body composition (p>0.05).

• Korean Journal of Food Science and Technology
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• v.28 no.5
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• pp.987-993
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• 1996

This study was attempted to prepare milk and soymilk containing high number of viable cells of bifidobacteria during the fermentation as well as to establish the optimum condition for bacteria growth. Activity of ${\alpha}$- and ${\beta}-galactosidase$ produced by bifidobacteria was also determined. Milk and soymilk inoculated with Bifidobacterium longum ATCC 15707 were incubated in a nitrogen-carbon dioxide atmosphere at $37^{\circ}C$ for two days. and time courses of pH, acidity, viable cells and effect of growth factors were determined. After two days, pH of milk gradually decreased from 6.81 to 4.84 and pH of soymilk changed from 7.02 to 3.89. The viable cell numbers of bifidobacteria increased constantly in soymilk, while bacterial growth in milk appeared to be delayed after storage of two days. Both of ${\alpha}$- and ${\beta}-galactosidase$ activities were detected in soymilk, but activity of ${\beta}-galactosidase$ was predominant in milk. Fucosyllactose appeared to be a good growth factor in soymilk. During the fermentation of milk, L-cysteine HCl enhanced growth of bifidobacteria at the early stage and fucosyllactose was a good growth factor in the propagations of bifidobacteria from middle stage.

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1430-1436
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• 2007

[ ${\alpha}$ ]-Galactosidase was immobilized in a mixture of k-carrageenan and locust bean gum. The properties of the free and immobilized enzyme were then determined. The optimum pH for both the soluble and immobilized enzyme was 4.8. The optimum temperature for the soluble enzymes was $50^{\circ}C$, whereas that for the immobilized enzyme was $55^{\circ}C$. The immobilized enzyme was used in batch, repeated batch, and continuous modes to degrade the raffinose-family sugars present in soymilk. Two hours of incubation with the free and immobilized ${\alpha}$-galactosidases resulted in an 80% and 68% reduction in the raffinose oligo saccharides in the soymilk, respectively. In the repeated batch, a 73% reduction was obtained in the fourth cycle. A fluidized bed reactor was also designed to treat soymilk continuously and the performance of the immobilized ${\alpha}$-galactosidase tested at different flow rates, resulting in a 90% reduction of raffinose-family oligosaccharides in the soymilk at a flow rate 40 ml/h. Therefore, the present study demonstrated that immobilized ${\alpha}$-galactosidase in a continuous mode is efficient for reducing the oligosaccharides present in soymilk, which may be of considerable interest for industrial application.

• Microbiology and Biotechnology Letters
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• v.39 no.3
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• pp.245-251
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• 2011

[ ${\alpha}$ ]Galactosidase was purified from a culture filtrate of Mortierella sp. by CM-sephadex C-50, and subsequent Sephadex G-100 column chromatography. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 56 kDa. $Gal^3Man^4$ ($6^3$-mono-O-${\alpha}$-D-galactopyranosyl-4-O-${\beta}$-D-mannotetraose), $Gal^{2,3}Man_5$ ($6^{2,3}$-di-O-${\alpha}$-D-galactopyranosyl-4-O-${\beta}$-D-mannopentaose), $Gal_2Man_3$ ($6^2$-mono-O-${\alpha}$-D-galactopyranosyl-4-O-${\beta}$-D-mannotriose), $Gal^2Man_6$ ($6^2$-mono-O-${\alpha}$-D-galactopyranosyl-4-O-${\beta}$-D-mannohexaose) and $Gal^2Man_5$ ($6^2$-mono-O-${\alpha}$-D-galactopyranosyl-4-O-${\beta}$-D-mannopentaose), prepared from 3 types of microbial ${\beta}$-mannnanase, were used as substrates. $Gal^3Man_4$ and $Gal^2Man_3$ had a stubbed ${\alpha}$-galactosyl residue on the $2^{nd}$ and $3^{rd}$ mannose from the reducing end of mannotetraose and mannotriose, thus ${\alpha}$-galactosidase showed a preference for stubbed ${\alpha}$-galactosyl residue. ${\alpha}$-Galactosidase hydrolyzed $Gal^3Man_4$ more rapidly than $Gal^2Man_3$. However, ${\alpha}$-galactosidase hardly acted on $Gal^{2,3}Man_5$, $Gal^2Man_6$ or $Gal^2Man_5$. The enzyme hydrolyzed melibiose to galactose and glucose, raffinose to galactose and sucrose, and also stachyose to galactose and raffinose.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.328-333
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• 2004

The maximum productivity of ${\alpha}-galactosidase,$ capable of hydrolyzing completely ${\alpha}-D-l,6-galactopyranosyl$ linkages within oligomeric substrates such as melibiose, raffinose and stachyose to liberate galactose residue, was reached to 718 mU/ml in the culture filtrate of Bacillus licheniformis at death phase. The ${\alpha}-galactosidase$ was identified to show different efficiencies for hydrolyzing the ${\alpha}-galactooligosaccharides$ according to analysis of reaction products by both TLC and quantification of the liberated reducing sugars. The enzyme was active on ${\alpha}-galactooligosaccharides$ in the order of melibiose, raffinose, and stachyose. Though the hydrolyzing activity of enzyme was faintly inhibited by reaction products such as galactose, glucose and sucrose with amounts of five folds more than the added substrates (20 mM), the largest inhibition of enzyme activity was caused by galactose among the end products. Unknown compound, which migrated slower than melibiose on TLC, was detected during hydrolysis reaction of melibiose, suggesting that the ${\alpha}-galactosidase$ has a glycosyl transferase activity. In addition, the enzyme was able to hydrolyze efficiently raffinose and stachyose existed in the soluble extract of soybean meal.

• 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 $50^\circ{C}$.