• Title/Summary/Keyword: Alpha-N -acetyl-D-glucosaminidase

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Isolation and Properties of $\beta$-N-Acetyl-D-glucosaminidase B from Rat Uterus

  • Jung, Jin-Ha;Yang, Chul-Hak
    • Bulletin of the Korean Chemical Society
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    • v.4 no.3
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    • pp.139-143
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    • 1983
  • ${\beta}$-N-Acetyl-D-glucosaminidase B was highly purified with the following sequence of steps; DEAE-cellulose, CM-cellulose, and Sephadex G-200 gel filtration chromatograpies. The specific activity of the purified ${\beta}$ -N-acetyl-D-glucosaminidase B was 2.2 units/mg protein with 12.9 % yield and 196.2 fold purity. The purified ${\beta}$-N-acetyl-D-glucosaminidase B showed single band on polyacrylamide gel electrophoresis. The final preparation of ${\beta}$ -N-acetyl-D-glucosaminidase B was completely free friom arylsulfatase and ${\beta}$-glucuronidase. ${\beta}$ -N-Acetyl-D-glucosaminidase B had pH optimum of 4.5 in 0.5 M sodium citrate buffer. The molecular weight of ${\beta}$-N-acetyl-D-glucosaminidase B was 133,000 by Sephadex G-200 gel filtration. The Km value of ${\beta}$-N-acetyl-D-glucosaminidase B using p-nitrophenyl-N-acetyl-${\beta}$-D-glucosaminide as substrate was 1.0 mM and $V_{max}$ was 0.014 ${\mu}$ mole/min. ${\beta}$-N-Acetyl-D-glucosaminidase B was stable at $55^{circ}C$ for 70 minutes. The crude ${\beta}$ -N-acetyl-D-glucosamiinidase in 70 % ammonium sulfate retained 93 % activity after 7 months storage at -$55^{circ}C$. Bovine serum albumin, sodium chloride, and phosphate activated ${\beta}$ -N-Acetyl-D-glucosaminidase B. N-Acetyl-D-glucosamine, ${\alpha}$-methyl-D-mannoside, and acetate inhibited ${\beta}$ -N-acetyl-D-glucosaminidase B.

Long-term clinical course of a patient with mucopolysaccharidosis type IIIB

  • Kim, Ja Hye;Chi, Yang Hyun;Kim, Gu-Hwan;Yoo, Han-Wook;Lee, Jun Hwa
    • Clinical and Experimental Pediatrics
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    • v.59 no.sup1
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    • pp.37-40
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    • 2016
  • Mucopolysaccharidosis type III (MPS III) is a rare genetic disorder caused by lysosomal storage of heparan sulfate. MPS IIIB results from a deficiency in the enzyme alpha-N-acetyl-D-glucosaminidase (NAGLU). Affected patients begin showing behavioral changes, progressive profound mental retardation, and severe disability from the age of 2 to 6 years. We report a patient with MPS IIIB with a long-term follow-up duration. He showed normal development until 3 years. Subsequently, he presented behavioral changes, sleep disturbance, and progressive motor dysfunction. He had been hospitalized owing to recurrent pneumonia and epilepsy with severe cognitive dysfunction. The patient had compound heterozygous c.1444C>T (p.R482W) and c.1675G>T (p.D559Y) variants of NAGLU. Considering that individuals with MPS IIIB have less prominent facial features and skeletal changes, evaluation of long-term clinical course is important for diagnosis. Although no effective therapies for MPS IIIB have been developed yet, early and accurate diagnosis can provide important information for family planning in families at risk of the disorder.

Seasonal Fluctuations of Heterotrophic Activity and Bacterial Extracellular Enzyme Activity in Paldang Lake (팔당호에서 종속영양 활성도의 계절적 변화 및 세균의 세포외 효소활성)

  • 김상진
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.93-98
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    • 1993
  • To investigate the organic matter transformation in aquatic environment, seasonal fluctuations of heterotrophic activity and microbia] extracellular enzyme activity were studied in Paldang Lake, Korea. The turnover time in the water column and the sediment at the station I fluctuated between 3 -I ,300 hrs and 17-170 hrs for glucose, 5 -1.900 hrs and 15-240 hrs for protein hydrolysate and 4-350 hrs and 15-230 hrs for acetic acid, respectively, indicating that the seasonal turnover time of organic substrates fluctuated drastically. The respiration ratios of glucose. protein hydrolysate and acetate were 23-32%, 38-41% and 22-28% in the water column and 34%, 61% and 41% in the sediment. respectively. These results showed that the respiration ratios in the sediment were higher than those in the water column regardless of kinds of organic substrates. The bacterial extracellular enzyme activities of $\alpha$-glucosidase. $\beta$-glucosidase, N-acetyl-$\beta$-D-glucosaminidase and aminopeptidase were 32-44%. 31-32%, 18-34% and 61-67% in the water column, and 34%. 40%, 23% and 65% in the sediment. respectively.

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Effects of Ginseng Saponin on the Lysosomal Enzyme Activities in Streptozotocin-induced Diabetic Mice (인삼 Saponin이 Lysosome 효소 활성에 미치는 영향)

  • 문창규;김희수;김강석
    • Journal of Food Hygiene and Safety
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    • v.9 no.3
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    • pp.105-109
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    • 1994
  • Lysosomal enzymes might play a most important role in the pathogenesis od diabetic microangiopathy. Some glycosidases, which participate in the catabolism of glycoprotein, are significantly decreased in diabetic mice. In search of new potential lysosomal enzyme inducers, we examined the effects of crude red-ginseng saponin fraction on N-acetyl-$\beta$-D-glucosaminidase, $\beta$-D-galactosidase and $\alpha$-D-mannosidase activities in the liver and kidney of normal and streptozotocin induced diabetic mice. It was found that i.p. administration of ginseng saponin produced the induction of lysosomal enzymes in the kidney more intensively than in the liver. The obtained results suggest the possibility that ginseng saponin might prevent the diabetic microangiopathy.

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Effects of the Acute and Subacute Administration of 1-(N-methyl) piperazinyl-3-phenyl-isoquinoline on Rat Kidney

  • Lim, Dong-Koo;Park, Sun-Hee;Noh, Eun-Young;Kim, Han-Soo;Cho, Won-Jea
    • Toxicological Research
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    • v.16 no.1
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    • pp.47-52
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    • 2000
  • To evaluate the renal toxicity of the antitumor agent, 1-(N-methyl) piperazinyl-3-phenyl-isoquinoline(CWJ-$\alpha$-5), rats were terated with CWJ-$\alpha$-5 (acute : 100mg/kg, i.p., single and subacute : 10mg/kr, i.p., daily for 7 days). The changes in the body weights, water consumption, kidney weights and urine volume after and during the treatment were observed. The concentrations of urinary creatinine, the activities of N-acetyl-$\beta$-D-glucosaminidase (NAG), alanine aminopeptidase (AAP), $\gamma$-glutamyl transpeptidase ($\gamma$-GT) and lactate dehydrogenase (LDH) in 24 hr urine were also determined. The body weight and water consumption were decreased after the acute and subacute administration. However, the excretion of urine was not changed except the 1 day after the acute treatment. The excretion of creatinine was significantly decreased from 1 day after acute administration and continuously decreased. Also the excretion of creatinine was decreased during subacute administration. However, the protein excretion did not changed in both treatment. Those indicate that CWJ-$\alpha$-5 might decrease the metabolic rate of muscle. The urinary activities of NAG, AAP, $\gamma$-GT, and LDH were significantly affected by the drug treatment. The urinary activities of NAG, AAP and $\gamma$-GT were significantly increased 1 and 3 days after the acute administration and then returned to the control value. However, the urinary activities of LDH were increased 7 days after acute treatment. During subacute treatment, the urinary activities of $\gamma$-GT were not changed. However, the urinary activities of NAG, AAP and LDH were only significantly increased after the third administration. These results indicate that either the high acute dose or the subacute administration with low dose of the compound might induce a temporal damage in the kidney cells.

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Changes of Glycosidase Activity of Frozen-Thawed Spermatozoa in Human

  • Lee, Chae-Sik;Lee, Sang-Chan;Lee, Ji-Eun;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.185-190
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    • 2011
  • To evaluate the effect of spermatozoa culture on glycosidase activity of frozen-thawed spermatozoa in human, 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 ${\beta}$-GlcNAc'ase activity was at least two-folds higher than other glycosidases regardless of spermatozoa incubation (p<0.05). The spermatozoa motility was decreased with incubation periods, but no effects by different glycosidases on the changes of spermatozoa motility during the various periods of incubation. In all glycosidases, the spermatozoa-zona binding rates in spermatozoa without incubation were higher than in spermatozoa incubated for 2 h (p<0.05). ${\beta}$-GlcNAc'ase is present mainly in the plasma membrane of spermatozoa frozen-thawed in human. It was also shown that the glycosidase activity was increased in all glycosidases in spite of lower sperm-zona binding by spermatozoa incubation.

돼지 동결정액의 배양에 따른 체외수정능력과 Glycosidase Activity의 변화

  • 황인선;정희태;양부근;김정익;박춘근
    • 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

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

  • 황인선;정희태;양부근;김정익;박춘근
    • 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.

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.
    • 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.

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Evaluation of the Activities of Antioxidant Enzyme and Lysosomal Enzymes of the Longissimus dorsi Muscle from Hanwoo (Korean Cattle) in Various Freezing Conditions

  • Kang, Sun Moon;Kang, Geunho;Seong, Pil-Nam;Park, Beomyoung;Kim, Donghun;Cho, Soohyun
    • Food Science of Animal Resources
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    • v.34 no.6
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    • pp.742-748
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    • 2014
  • This study was conducted to evaluate the activities of antioxidant enzyme (glutathione peroxidase (GSH-Px)) and lysosomal enzymes (alpha-glucopyranosidase (AGP) and beta-N-acetyl-glucosaminidase (BNAG)) of the longissimus dorsi (LD) muscle from Hanwoo (Korean cattle) in three freezing conditions. Following freezing at -20, -60, and $-196^{\circ}C$ (liquid nitrogen), LD samples (48 h post-slaughter) were treated as follows: 1) freezing for 14 d, 2) 1 to 4 freeze-thaw cycles (2 d of freezing in each cycle), and 3) refrigeration ($4^{\circ}C$) for 7 d after 7 d of freezing. The control was the fresh (non-frozen) LD. Freezing treatment at all temperatures significantly (p<0.05) increased the activities of GSH-Px, AGP, and BNAG. The $-196^{\circ}C$ freezing had similar effects to the $-20^{\circ}C$ and $-60^{\circ}C$ freezing. Higher (p<0.05) enzymes activities were sustained in frozen LD even after 4 freeze-thaw cycles and even for 7 d of refrigeration after freezing. These findings suggest that freezing has remarkable effects on the activities of antioxidant enzyme and lysosomal enzymes of Hanwoo beef in any condition.