• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.25-25
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• 2003

본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity의 관계를 검토하였으며, 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glycosidase 특이성을 확인하기 위하여 $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase 및 N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase)의 activity를 분석하였다. 그 결과 glycosidase activity는 동결정자의 융해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity는 정자 배양 유무에 관계없이 다른 glycosidase 처리시보다 최소한 2배 이상 높게 나타났다. 또한 첨체반응이 유기된 정자의 비율은 glycosidase ($\alpha$-D-mannosidase; P<0.05)에 의해 영향을 받았으며 정자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 생존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 침입에 대한 또 다른 실험에서, 서로 다른 glycosidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다($\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착 정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase첨가시 나타났다. 또한 모든 glycosidase 처리시 2시간 배양한 정자보다는 배양하지 않은 정자에서 투명대에 대한 접착정도가 높게 나타났으며, $\alpha$-D-mannosidase의 처리시 유의적인 차이를 보였다(P<0.05). 본 연구의 결과, $\beta$-GlcNAc'ase가 주로 돼지정자의 원형질막내에 존재하는 것으로 추측되며, 배양된 정자에 의한 투명대 접착정도와 침입율이 낮았음에도 불구하고 glycosidase activity가 증가하는 것으로 나타났다

• Journal of Sericultural and Entomological Science
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• v.39 no.1
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• pp.86-92
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• 1997

The effects of silkworm extract on body weight, blood glucose and insulin levels, and an intestinal glycosidase activity were studied in high carbohydrate diet administered mice. Hyperglycemia and hyperinsulinemia represented in high carbohydrate diet administered mice disappeared when silkworm extract(50 mg) was added to 100 g diet for 10 week period. These results suggest that silkworm extract may resolve the insulin resistance by lowering serum insulin level resulting from inhibition of intestinal glycosidase activity. Silkworm extract coadministered with high carbohydrate diet to mice for 10 weeks significantly induced maltase, sucrase and lactase acivities, especially middle and distal portion. Further studies about regulatory mechanisms of glycosidase by silkworm extract are needed.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.638-642
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• 2000

A ${\beta}-glycosidase$ enzyme with $\beta$-D-fucosidase, ${\beta}-D-galactosidase$, and $\beta$-D-glucosidase activities has been purified from Thermus caldophilus GK24. The enzyme was monomeric with a molecular mass of 49 kDa, as evidenced by SDS-PAGE. The $K_m$ values for p-nitrophenyl ${\beta}-D-fucopyranoside$ (p-NPFuc), p-nitrophenyl ${\beta}-D-galactopyranoside$ (p-NPGal), and p-nitrophenyl ${\beta}-D-glucopyranoside$ (p-NPGlu) were 0.23 mM, 6.25 mM, and 0.28 mM, respectively. The enzyme showed optimal pH ranging between 5.5-6.5 and maximum temperature in the range of $85-90^{\circ}C$ for all the above mentioned activities. The half-life of the enzyme in sodium phosphate buffer (pH 6.0) at $80^{\circ}C$ was approximately 7 h. The p-NPGal hydrolyzing activity of Tca ${\beta}-glycosidase$ was strongly activated by L-histidine, while the p-NPFuc and p-NPGlu hydrolyzing activities of Tca ${\beta}-glycosidase$ were not affected at all by the amino acid. These results suggest differences in the conformation or in the reactive residues at the active site of Tca ${\beta}-glycosidase$.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.43-43
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• 2001

This study evaluated the effect of fertilization-promoting peptide (FPP) on fertilizing ability and glycosidase activity in vitro of spermatozoa frozen-thawed in pig, Using chlortetracycline fluorescence analysis, the various glycosidase analyses and the oocyte penetration test, we have obtained evidence that FPP can promote the fertilizing ability and glycosidase activity of frozen-thawed spermatozoa in vitro. When frozen-thawed spermatozoa was washed with different concentrations of FPP, there were significantly (P<0.05) more acrosome-reacted in medium with 100 nM than 0, 50, 200 and 400 nM. The penetration rates were also highest in medium containing with 100 nM FPP (P<0.05). On the other hand, the $\beta$-N-acetylglucosaminidase activity was at least twofold higher than other glycosidase. In same glycosidase, however, there were no difference in medium with different concentrations of FPP In another experiment, spermatozoa preincubated in medium with or without FPP for 0, 1, 2, 3 and 4 h were inseminated with oocytes matured in vitro. The percentages of spermatozoa that reached acrosome reaction were affected by preincubation and were higher in medium with that than without FPP. When oocytes were inseminated with spermatozoa preincubated in medium with and without FPP during the different periods, however, penetration rates were decreased with preincubation periods of spermatozoa. On the other hand, when the sperm-oocyte were cultured for 4, 8, 12, 16, 20 and 24 h, the penetration rates were higher in spermatozoa preincubated with that than without FPP and had a tendency to increase as time of culture periods. However, The activities of $\alpha$-fucosidase, $\alpha$ -mannosidase, $\beta$-galactosidase and N-acetyl- $\beta$-D-glucosaminidase were higher in medium with that than without FPP regardless of periods of sperm preincubation and sperm-oocyte culture. These results suggest that FPP may play a positive role in promoting of sperm function and glycosidase activity in vitro in pig.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.153-161
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• 2003

This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase and N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The $\beta$-GlcNAc'ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc'ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc'ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with $\alpha$-D-mannosidase (P<0.05). These results suggest that $\beta$-GlcNAc'ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.

• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.151-156
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• 2002

This study was performed to obtain the thermostable $\beta$-glycosidase producing bacteria from hot spring of volcanic area at Atagawa in Japan. KNOUC 202 was selected because it showed thermostable $\beta$-glycosidase activity in sodium phosphate buffer(pH 6.8) at $70^{\circ}C$ for 4h, and it was identified. The strain was aerobic, asporogenic bacilli, immobile, gram negative, catalase positive, oxidase positive, and pigment-producing. Optimum growth was at $70~72^{\circ}C$, pH 7.0~7.2, and it could grow in the presence of 3% NaCl. The main fatty acids in cell were iso-15:0 and iso-l7:0. 16S rRNA sequence of KNOUC 202 showed 99.9% similarity with that of Thermus thermophilus ATCC 27634(HB8). Based on morphological, physiological, biochemical characteristics, cellular fatty acids profile and 16S rRNA sequence analysis, KNOUC 202 was identified as Thermus thermophilus.

• Biomolecules & Therapeutics
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• v.6 no.4
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• pp.364-370
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• 1998

Glycosidase inhibitors from urine of Bombyx mori were isolated and their inhibitory activities on glycosidases were evaluated. Six compounds were isolated by using several ion exchange columns, and their chemical structures were identified by the physicochemical and spectral data. Compound IV, V and Ⅵ were identified as 1-deoxynojirimycin, fagomine and 1,4-dideoxy-1,4-imino-D-arabinitol, respectively. Among six compounds isolated,1-deoxynojirimycin(IV) was the most potent inhibitor on $\alpha$-glucosidase and $\beta$-galactosidase of rat intestine, and its inhibitory activities for trehalase and almond $\beta$-glucosidase were relatively weak. Compound V and Ⅵl retained a little inhibitory potency toward $\alpha$-glucosidase and $\beta$-galactosidase. Compound II and III, however, have been found to have no effect on all glycosidases tested in this study.

• Biomolecules & Therapeutics
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• v.6 no.3
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• pp.242-246
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• 1998

The inhibitory activities of Amberlite active fraction, which was obtained from methanol soluble fraction of freeze dried slikworm urine, on the rat intestinal glycosidase-catalyzed enzymatic reaction were examined in in viro and in vivo experiments. Amberlite active fraction showed significant inhibitory effects on the hydrolysis of o-glycosidic bond, especially $\alpha$-1,4 bond. On the other hand, the inhibition on the hydrolysis of $\beta$-glycosidic bond was very weak. Oral administration of Amberlite active fraction resulted in a dose-dependent decrease in the blood glucose after an oral maltose load, and postprandial hyperglycemia in carbohydrate-loaded mice was suppressed by Amberlite active fraction at 60 mgHg in decreasing order of maltose, starch, sucrose and lactose. 60 mg/kg of Amberlite active fraction lowered the blood glucose level markedly after 18, 35, and 60 min after an oral maltose load and the antihyperglycemic activity was maintained upto 90 min. In alloxan-induced hyperglycemic mice, Amberlite active fraction at a dose of 100 mg/kg also significantly lowered blood glucose after an oral maltose load, and its efficacy was almost equivalent to that of acarbowe.

• YAKHAK HOEJI
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• v.41 no.4
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• pp.484-491
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• 1997

The effect of prolonged treatment with Mori Folium column fraction(MFCF) on intestinal ${\alpha}$-glycosidase activity has been studied in mice made hyperglycemic and hyperinsulinemic by feeding a high carbohydrate-containing diet. Mice were treated for 10 week with or without MFCF, added to the high carbohydrate-containing diet at 50mg/100g food. While MFCF had no effect on body weight, it prevented the rise in glycemia and insulinemia. Maltase, sucrase and lactase activities were measured in intestinal homogenates of proximal, middle and distal segments of jejunoileum. Following 10 week of MFCF administration, MFCF significantly induced all three enzyme activities especially at middle and distal segments. The mechanisms responsible for these changes and their potential biochemical implications remain to be determined.

• Proceedings of the Korean Society of Life Science Conference
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• 2007.10a
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• pp.54.2-54.2
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• 2007

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