• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.800-808
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• 2005

Leuconostoc mesenteroides SY1, an isolate from kimchi, was able to ferment $\alpha$-galactosides, such as melibiose and raffinose. $\alpha$-Galactosidase ($\alpha$-Gal) activity was higher in cells grown on melibiose and raffinose than cells grown on galactose, sucrose, and fructose. $\alpha$-Gal activity was not detected in cells grown on glucose, indicating the operation of carbon catabolite repression (CCR). A 6 kb DNA fragment was PCR amplified using a primer set based on the nucleotide sequence of a putative $\alpha$-galactosidase gene (aga) from L. mesenteroides ATCC 8293. Nucleotide sequencing of the 6 kb fragment confirmed the presence of aga and other genes involved in the galactosides utilization, and the gene order was galR (transcriptional regulator)-aga-gaIK (galactokinase)-gaIT (galactose-1-phosphate uridylyltransferase). Northern blotting experiment showed that aga, gaIK, and gaIT constituted the same operon, that the transcription was induced by galactosides, such as melibiose and raffinose, whereas gaIR was independently transcribed as a monocistronic gene, and that the level of transcription was fairly constant. The aga was overexpressed in E. coli BL21 (DE3) using pET26b(+) vector, and $\alpha$-Gal was accumulated in E. coli as an inclusion body.

• Journal of Plant Biology
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• v.27 no.2
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• pp.43-50
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• 1984

Effects of IAA on the elongation and cell wall hlysocidase activities were investigated in excised rape (Brassica napus L. cv. Yongdang) hypocotyl segments. IAA promoted the elongation of rape hypocotyl segments. In rape hypocotyls, the first 10-mm segments from the hook exhibited maximal elongation and the capacity of elongation was gradually decreased with increasing distance of each 10-mm from the hook. A good correlation has been obtained between the magnitude of endogenous growth and the activities of $\alpha$, $\beta$-glucosidase and $\alpha$, $\beta$-galactosidase. However, exogenous application of IAA did not seem to enhance the tissue with IAA resulted in acidification of the incubation medium. From these data, we can conclude that IAA seems to enhance elongation of the tissue segments, at least in part, by releasing hydrogen ion into cell wall, some of which may participate in the cell wall extension process, but does not seem to trigger the activation of $\alpha$, $\beta$-glucosidase and $\alpha$, $\beta$-galactosidase.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• Journal of Animal Science and Technology
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• v.45 no.2
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• pp.211-218
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• 2003

For the Exp. 1, a total of sixty pigs (10.57$\pm$0.30kg average initial body weight) were used in a 15-d growth assay to determine the effect of dietary $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase on growth performance and nutrient digestibility. Dietary treatments included 1) CON (corn-dried whey-SBM based diet), 2) EC0.1 (CON diet+0.1% enzyme complex of $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase). Through the entire experimental period, gain/feed of pigs fed EC0.1 diet was higher (0.43 vs 0.52) than that of pigs fed CON diet (P<0.05). Pigs fed EC0.1 diet showed significant (P<0.05) improvement in dry matter (74.82% vs 82.41%) and nitrogen (70.59% vs 77.88%) digestibilities compared to pigs fed CON diet. For the Exp. 2, a total of thirty six pigs (22.30$\pm$0.45kg average initial body weight) were used in a 30-d growth assay to determine the effects of dietary $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase in low energy diet on growth performance and nutrient digestibility. Dietary treatments included 1) AME (adequate ME diet), 2) AME+EC0.1 (AME diet+0.1% enzyme complex) and LME+EC0.1 (low ME diet + 0.1% enzyme complex). Through the entire experimental period, average daily feed intake of pigs fed enzyme complex supplemented diets was higher than that of pigs fed CON diet (P<0.05). Also, pigs fed AME+EC0.1 diet showed significant (P<0.05) increase in ADFI (1,401g vs 1,733g) compared to pigs fed CON diet. Pigs fed enzyme complex supplemented diet showed significant (P<0.05) improvement in dry matter and nitrogen digestibilities compared to pigs fed CON diet. In conclusion, the results obtained from these feeding trials suggest that the supplementation of $\alpha$-1,6-galactosidase and $\beta$-1,4-mannanase was an effective means for improving growth performance and dry matter and nitrogen digestibilities in nursery and growing pigs.

• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.14 no.2
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• pp.135-141
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• 2014

Objectives: Fabry disease (FD) is a lysosomal storage disease caused by the inappropriate accumulation of globotriaosylceramide (Gb3) in tissues due to a deficiency in the enzyme ${\alpha}$-galactosidase A. Hypertrophic cardiomyopathy is one of the chronic complications of FD. We tried to evaluate the prevalence of Fabry disease in the Korean patients with left ventricular hypertrophy (LVH). Methods: A total of 257 patients with LVH were recruited and they were 172 males (mean 56 years, range 30-81 years) and 84 females (mean 66 years, range 45-85 years). Urinary Gb3 was used to screen FD by high performance liquid chromatography-tandem mass spectrometry. Confirmatory tests were done by alpha-galactosidaseA activity using fluorometric assay and by GLA mutation analysis using sequencing. Results: Four patients were screening positive by urinary Gb3 analysis (cutoff, 25 ug/mmol creatinine). But, one female patient was diagnosed with FD confirmed by enzyme analysis in leukocytes as well as by genetic analysis (1/257 patients, 0.4%). She showed 54.3 ug/mmoL creatinine of Gb3 and 15.5 nmole/hr/mg protein (reference range, $55.2{\pm}12.7nmole/hr/mg$ protein) of alphagalactosidase A activity. And she had a heterozygous GLA mutation of c.796G>A (p.D266N). Her daughter was found to be a carrier for FD confirmed by GLA mutation analysis. Asymptomatic carrier showed 25.5ug/mmol creatinine of Gb3 and 42.5 nmole/hr/mg protein (reference range, $55.2{\pm}12.7nmole/hr/mg$ protein) of alpha-galactosidase A activity. Conclusions: The prevalence of FD in Koran patients with LVH was detected as 0.4%. Although the prevalence seems to be low, screening studies are of great importance for detecting hidden cases as well as for identifying other effected family members.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.134-134
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• 1998

The modulation of glycosidase activity by inhibitors is of great interest. Such compounds have been shown to be important tools in mechanistic studies on glycohydrolase as well as having promising therapeutic application. An ${\alpha}$-glucosidase inhibitor was isolated from culture filterates of Penicillium sp. The inhibitor was active against ${\alpha}$-glucosidase isolated from yeast and porcine small intestine. However, it showed no inhibition to Aspergillus ${\alpha}$-galactosidase, Escherichia coli ${\beta}$-galactosidase, and jack bean ${\alpha}$-mannosidase. The inhibitor was highly soluble in ether, methanol and chloroform. The inhibitor was purified using silica gel, Sephadex LH-20 column chromatography and reverse-phase HPLC. The inhibitory compound designated PA-7(IC$\sub$50/=35$\mu\textrm{g}$) was obtained as white powder. The structure of PA-7 was determined with spectroscopic data of EI-MS, FAB-MS, $^1$H, and $\^$13/C NMR. The inhibitor has a diketopiperazine moiety.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1340-1347
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• 2010

To study the effects of ${\alpha}$-galactosidase (${\alpha}$-Gal) supplementation on performance and energy metabolism, 216 Arbor Acres male broilers were placed in 36 cages of 6 birds each and allotted to 4 diets for 42 d, with 0-21 d as starter period and 22-42 d as grower period. The 4 diets were based on corn non-dehulled soybean meal in a $2{\times}2$ factorial arrangement, with 2 levels of ${\alpha}$-Gal (0 vs. 60 U/kg feed) and 2 levels of ME (normal metabolizable energy (NME) and low metabolizable energy (LME)). Bird performance was obtained at 21 and 42 d of age with samples of feces collected for nutrient digestibility from 19-21 d and 40-42 d. At 21 and 42 d, 1 bird from 6 cages of each treatment was killed to determine liver weight, intestinal pH and chyme viscosity. With the addition of ${\alpha}$-Gal the 42 d body weight (BW) and 0-42 d average daily gain (ADG) were significantly improved (p<0.05). Average daily feed intake (ADFI) of birds fed the LME diet was significantly increased compared to those fed the NME diet during starter (p<0.01) and grower (p<0.05) periods and overall (p<0.01). There was an interaction of ${\alpha}-Gal{\times}ME$ on 0-21 d ADFI (p<0.01). Supplementation of ${\alpha}$-Gal significantly improved (p<0.01) feed efficiency during the grower period and overall. Feed efficiency of birds fed the LME diet was significantly decreased (p<0.05) compared to those fed the NME diet during the starter period and overall. With the addition of ${\alpha}$-Gal apparent metabolizable energy (AME) was improved (p<0.01) by 2.1% and 1.8% during starter and grower periods, respectively. There was a main effect (p<0.05) of ${\alpha}$-Gal on the digestion of neutral detergent fiber (NDF) during the starter period and crude protein (CP), NDF and acid detergent fiber (ADF) during the grower period. With the addition of ${\alpha}$-Gal, the relative weight of liver was reduced (p<0.01) during the two phases. The duodenal and jejunal pH were significantly decreased (p<0.01) with the supplementation of ${\alpha}$at the two phases. ${\alpha}$-Gal addition reduced (p<0.01) chyme viscosity of the ileum during the starter and grower periods. Overall, ${\alpha}$-Gal showed a major effect on nutrient efficiency, improved ADG and feed efficiency, whereas LME decreased feed efficiency. The incorporation of ${\alpha}$-Gal into a LME diet could at least partially offset ME deficiency of non-dehulled soybean meal.

• BMB Reports
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• v.46 no.3
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• pp.157-162
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• 2013

Human ${\alpha}$-galactosidase A (GLA) has been used in enzyme replacement therapy for patients with Fabry disease. We expressed recombinant GLA from Chinese hamster ovary cells with very high productivity. When compared to an approved GLA (agalsidase beta), its size and charge were found to be smaller and more neutral. These differences resulted from the lack of terminal sialic acids playing essential roles in the serum half-life and proper tissue targeting. Because a simple sialylation reaction was not enough to increase the sialic acid content, a combined reaction using galactosyltransferase, sialyltransferase, and their sugar substrates at the same time was developed and optimized to reduce the incubation time. The product generated by this reaction had nearly the same size, isoelectric points, and sialic acid content as agalsidase beta. Furthermore, it had better in vivo efficacy to degrade the accumulated globotriaosylceramide in target organs of Fabry mice compared to an unmodified version.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.