• 제목/요약/키워드: glycosidase

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돼지 동결정액의 배양에 따른 체외수정능력과 Glycosidase Activity의 변화

  • 황인선;정희태;양부근;김정익;박춘근
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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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

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고탄수화물 식이투여 마우스에서 누에추출물이 소장내의 $\alpha$-glycosidase 활성에 미치는 영향 (Effect of Silkworm Extract on Intestinal $\alpha$-Glycosidase Activity in Mice Administered with a High Carbohydrate-containing Diet)

  • 정성현;김미선;류강선
    • 한국잠사곤충학회지
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    • 제39권1호
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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.

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Purification and Characterization of a Thermostable ${\beta}-Glycosidase$ from Thermus caldophilus GK24

  • Yoo, Jin-Sang;Han, Ki-Woong;Kim, Hyun-Kyu;Kim, Min-Hong;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • 제10권5호
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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 ${\beta}-glycosidase$.

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The Effect of Fertilization-Promoting Peptide on Fertilizing Ability and Glycosidase Activity In Vitro of Frozen-Thawed Boar Spermatozoa

  • Hwang, I.S.;Park, C.K.;H.T. Cheong;B.K. Yang;Kim, C.I.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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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.

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돼지 동결정액의 배양에 따른 체외수정능력과 Glycosidase Activity의 변화 (Changes of Glycosidase Activity and Fertilizing Ability in Vitro by Incubation of Frozen-Thawed Spermatozoa in the Pig)

  • 황인선;정희태;양부근;김정익;박춘근
    • 한국가축번식학회지
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    • 제27권2호
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    • pp.153-161
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    • 2003
  • 본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity를 평가하기 위하여 chlortetracycfine fluorescence분석법을 이용하여, glycosidase가 체외수정능력과 정자침입에 미치는 영향에 대하여 검토하였다. 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glyco-sidase 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 응해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity 는 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 자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 입에 대한 또 다른 실험에서, 서로 다른 glyco-sidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다 $\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase 첨가시 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.

일본의 Atagawa 온천지대에서 분리한 내열성 \beta-glycosidase 생성균주의 분리 및 동정 (Isolation and Identification of Thermostable \beta-glycosidase-producing Microorganism from Hot Spring of Volcanic Area at Atagawa in Japan.)

  • 남은숙;최종우;차성관;안종건
    • 한국미생물·생명공학회지
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    • 제30권2호
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    • pp.151-156
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    • 2002
  • 내열성 $\beta$-glycosidase 효소를 생성하는 균주를 분리하기 위해서 일본의 Atagawa 온천지역에서 시료를 채취하였다. 조효소액이 $70^{\circ}C$에서 4시간 동안 $\beta$-glycosidase활성을 유지하는 KNOUC 202를 선발하여 분리, 동정하였다. 선발된 KNOUC 202는 호기성으로 그람음성 간균이며, 포자를 형성하지 않으며, 운동성이 없으며, carotenoid를 생성하고, catalase, oxidase 양성으로 나타났으며, 최적 성장온도는 $70~72^{\circ}C$이고, 최적 pH는 7.0~7.2이었으며, NaCl 3% 농도에서 성장하였다. 세포내 지방산의 주성분은 iso-15:0과 iso-17:0이었다. KNOUC 202의 16S rRNA sequence는 Thermus thermophilus ATCC 27635(HB8)와 유사도가 99.9% 이었다. 따라서 형태학적, 이화학적, 생화학적 특성, 지방산조성 및 16S rRNA sequence 분석결과에 근거하여 KNOUC 202를 Thermus thermophilus로 동정하였다.

잠뇨로부터 질소함유 당물질 분리 및 glycosidase에 대한 저해활성 (Isolation of N-Containing Sugars from Silkworm Urine and Their Glycosidase Inhibitory Activities)

  • 송주경;정성현
    • Biomolecules & Therapeutics
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    • 제6권4호
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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.

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잠뇨의 흰쥐 소장내 glycosidase 저해활성 (Inhibitory Effect of Silkworm Urine on the Rat Intestinal Glycosidase)

  • 송주경;정성현
    • Biomolecules & Therapeutics
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    • 제6권3호
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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.

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고탄수화물 식이 투여 마우스에서 상엽 컬럼분획물의 ${\alpha}$-glycosidase 활성에 미치는 영향 (Effect of Mori Folium Column Fraction on Intestinal ${\alpha}$-glycosidase Activity in Mice Administered with a High Carbohydrate-Containing Diet)

  • 정성현;김미선;조여원
    • 약학회지
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    • 제41권4호
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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.

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