• 제목/요약/키워드: $\beta$-N-acetyl 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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    • 제4권3호
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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.

Testosterone과 dibutyryl cyclic AMP가 거세한 흰쥐 부정소의 $\beta$ -glucosidase와 몇가지 glycosidase 활성에 미치는 영향 및 부정소 상피세포의 여러 유형에 관한 연구 (Studies on Adivitie of $\beta$-Glucuronidase and Several Glycosidases of the Castrated Rat Epi-didymis Treated with Testosterone and Dibutyryl cAMP and the Cell Types of Epididymal Epithelium)

  • 최임순;정경순
    • 한국동물학회지
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    • 제32권3호
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    • pp.290-303
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    • 1989
  • $\beta$-glucosidase, $\beta$-glucuronidase, N-acetyl-$\beta$-glucosaminidase의 성적 성숙과의 연관서을 조사하기 위하여 흰쥐의 복강내로 testosterone과 dibutryjry cyclic AMP 를 투여하여 위이 효소들의 활성도를 측정하였다. 그 결과 겨세한 실험군에서$\beta$-glucosidase와 N-acetyl-$\beta$-glucosaminidase이 활성도도 거세후 7일째에는 유의성있는 감소효과를 나타내었다. testosterone을 7일간 계속 투여한 경우에는 세 효소의 활성도가 모두 유의성있게 증가하였고 dbcAMP 투여군의 경우는 거세 14일째되는 실험군과 비슷하거나 감소하는 경향이 있었다. 미세구조를 관찰한 결과 부정소 상피세포의 유형은 크게 주세포와 기저세포로 나눌 수 있었으며 주세포는 일반적인 원주상피의 형태를 나타냈으며 소낭을 많이 포함하고 있는 narrow cell이 존재하였다. 특히 부정소미에는 다른 상피세포에 비해 전자밀도가 낮은 light cell이 존재했고 기저세포는 부정소 부위마다 비슷한 형태를 가지는 것으로 나타났으며 상피세포 사이에는 이동능력을 가진 halo cell이 존재했다.

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한국인 분변으로부터 분리된 Bacteroides fragilis Roid 8의 Glycosidase 패턴 (Glycosidase Pattern of Bacteroides fragilis Roid 8 Isolated from a Korean Adult Feces)

  • 지근억;이세경
    • 한국식품과학회지
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    • 제25권2호
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    • pp.191-195
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    • 1993
  • 인체의 대장은 여러 종류의 균들에 의하여 상재균총이 이루어져 있는데 이들중 혐기성 균들이 주종을 이루고 있다. 이들 혐기성 균들 중 가장 많은 수가 Bacteroides이다. 본 연구에서 한국인으로부터 분리된 Bacteroides fragilis Roid 8은 장내의 다른 혐기성 균주들에 비하여 $N-acetyl-{\beta}-glucosaminidase$, ${\alpha}-fucosidase$, ${\beta}glucuronidase$ chitobiase, PNPCase 등의 활성이 높았다. ${\beta}-galactosidase$, ${\beta}-xylosidase$, ${\alpha}-arabinofuranosidae$활성은 없었고 ${\alpha}-glucosidase$, ${\beta}-glucosidase$, ${\alpha}-galactosidae$ 등의 생산은 Bifidobacteria 에 비하여 낮았다. BHI 기본배지에 여러 종류의 탄수화물을 첨가하여 배양한 뒤 생산된 $N-acetyl-{\beta}-glucosaminidae$, ${\alpha}-fucosidase$, ${\beta}-glucuronidase$ chitobiase, PNPCase, ${\beta}-glucosidase$, ${\beta}-glucosidase$ ${\alpha}-galactosidase$ 활성을 조사한 결과 모두 glucose와 lactose 첨가배지에서 이들 효소들의 활성이 낮았다. 조사된 모든 효소들에 대하여 특이적으로 현저히 생산을 증가시키는 당은 없었다. 이들 8개의 효소에 대하여 최적 pH와 최적온도가 조사되었다.

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N-acetyl-$\beta$-D-glucosaminidase 활성치에 의한 젖소의 준임상형유방염의 진단과 염산 Levamisole의 치료시험 (Diagnosis of Bovine Subclinical Mastitis by the Measurement of the N-acetyl-$\beta$-D-glucosaminidase Activity and Effect of Levamisole Treatment)

  • 강병규;양건묵
    • 한국임상수의학회지
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    • 제5권1호
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    • pp.23-30
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    • 1988
  • A total of 127 foremilk samples from dairy farms in Chonnam district was examined for the subclinical mastitis eve. six months, using a method of the N-acetyl-${\beta}$ -D-glucosaminidase (NAGase) test in relation to the California mastitis test(CMT) and the somatic tell count(SCC) and the compatibility and efficiency rating between the NAGase test and the other screening test were conducted. Fifteen subclinically mastitic cows were treated with a single oral dose of 7.5mg/kg of levamisole hydrochloride. The results are summarized as follows. 1. A linear relationship was found among the NAGase level, the CMT score and the SCC level, and it was found that NAGase activity measurements were comparable with other screening tests for diagnosing cows with mastitis. 2. Compatibilites between the NAGase and the CMT were 96.1%, and that of the NAGase and SCC were 93.7%. On the other hand, relative efficiency ratings of Postle's equation between the NAGase and CMT were 89.4%, and that of the NAGase and SCC were 84.1%. 3. Considering the result of CMT, SCC and NAGase level after treatment with levamisole hydrochloride it seemed to be of special value in the treatment of bovine subclinical mastitis.

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모 공단지역 사무직 근로자들의 요중 N-acetyl-$\beta$-D-glucosaminidase 역가에 관한 연구 (A Study on Urinary N-acetyl-$\beta$-D-glucosaminidase Activities of Office Workers in a Certain Industrial Complex Area)

  • 김화성;리갑수;이성수;안규동;이병국
    • Journal of Preventive Medicine and Public Health
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    • 제27권3호
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    • pp.547-556
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    • 1994
  • In order to identify the necessary information of biochemical Indices for renal effect of lead for the early detection in medical surveillance of lead worker, the reference values of urinary N-acetyl-$\beta$-D-glucosaminidase (NAG) activities were studied with 205 office workers in one industrial complex area who were not exposed to lead occupationally. While study variables selected for lead exposure were blood lead (PbB), blood zinc protoporphyrin(ZPP) and $\delta$-aminolevulinic acid (DALA) in urine, those for renal effect were urinary N-acetyl-$\beta$-D-glucosaminidase (NAG), blood urea nitrogen (BUN), serum creatinine(Cr), serum uric acid (Ua), and urinary total protein(U-TP). The results obtained were as follows: 1. The mean values of blood lead, ZPP and DALA in all subjects were $14.39{\pm}4.02{\mu}g/dl,\;21.61{\pm}8.00{\mu}g/dl,\;and\;2.73{\pm}0.90mg/l$ respectively. 2. The mean value of urinary NAG activities in all subjects was $3.51{\pm}2.01U/l$. The mean value of urinary NAG activities, which calculated from NAG activities divided by urinary creatinine concentration (CNAG), was $5.42{\pm}5.53U/g$ creatinine and log-arithmic normal distributed. 3. The reference value of urinary NAG activity was 12.06 U/g creatinine(95% CU=10.57-14.76 U/g creatinine). 4. Logarithmic CNAG(r=0.781 p<0.0l), U-TP(r=0.670 p<0.01) and ZPP(r=0.172 p<0.05) showed statistically significant correlation with CNAG.

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Correlation between glomerular filtration rate and urinary N acetyl-beta-D glucosaminidase in children with persistent proteinuria in chronic glomerular disease

  • Hong, Jeong-Deok;Lim, In-Seok
    • Clinical and Experimental Pediatrics
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    • 제55권4호
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    • pp.136-142
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    • 2012
  • Purpose: Urinary excretion of N acetyl-beta-D glucosaminidase (NAG) and ${\beta}_2$-microglobulin (${\beta}_2$-M) was increased in the presence of proximal tubular damage. Based on these urinary materials, we investigated the ability of expecting renal function in chronic glomerular diseases. In this study, we evaluated the relationship between glomerular filtration rate (GFR) urinary NAG, and urinary ${\beta}_2$-M. Methods: We evaluated 52 children with chronic kidney disease at the Chung-Ang University Hospital between January 2003 and August 2009. We investigated the 24-hour urinalysis and hematologic values in all 52 patients. Serum creatinine, creatinine clearance (Ccr), serum cystatin C, urinary ${\beta}_2$-M and urinary NAG were measured. Results: Out of 52 patients, there were 13 children with minimal change in disease, 3 children with focal segmental glomerulosclerosis, 17 children with immunoglobulin A nephropathy, 15 children with Henoch-Sch$\ddot{o}$nlein purpua nephritis, 3 children with poststreptococcal glomerulonephritis, and 1 child with thin glomerular basement membrane disease. In these patients, there were significant correlation between the Ccr and urinary NAG (r=-0.817; $P$ <0.01), and between the GFR (as determined by Schwartz method) and urinary NAG (r=-0.821; $P$ <0.01). In addition, there was a significant correlation between the GFR (as determined by Bokencamp method) and urinary NAG (r=-0.858; $P$ <0.01). Conclusion: In our study, there was a significant correlation between the GFR and urinary NAG, but there was no correlation between the GFR and urinary ${\beta}_2$-M, suggesting that the GFR can be predicted by urinary NAG in patients with chronic glomerular disease.

Glycerol이 흰쥐 신장에서의 Malondialdehyde 함량과 Superoxide Dismutase 활성도 및 요중 단백질 배설량과 $N-acetyl-{\beta}-D-glucosaminidase$ 활성도에 미치는 영향 (Effects of Glycerol on the Malondialdehyde Level and Superoxide Dismutase Activity in the Kidney and Urinary Protein Excretion and $N-acetyl-{\beta}-D-glucosaminidase$ Activity of the Rats)

  • 신인철;고현철
    • 대한약리학회지
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    • 제32권2호
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    • pp.259-267
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    • 1996
  • In an attempt to dofine the early biochemical determinants that participate in the pathogenesis of glycerol-induced nephrotoxicity, especially focusing on oxygen free radicals and $N-acetyl-{\beta}-D-glucosaminidase$ (NAG) activity, we studied 24-hours urine outflow, 24-hours urinary protein excretion and urinary NAG activity after the injection of glycerol and also we studied malondialdehyde(MDA) level and superoxide dismutase(SOD) activity in the kidney of rats at 24hr after the injection of glycerol. Sprague-Dawley albino rats weighing 240 to 260 gm were injected intramuscularly with a 50% solution of glycerol(2ml/kg, 4ml/kg and 8ml/kg). The group treated with glycerol showed significantly lower urine outflow level and urinary protein excretion level and higher urinary NAG activity after the injection as compared to those of control group. Also the group treated with glycerol showed significantly higher MDA level and lower SOD activity at 24hr after the injection as compared to those of control group. These results suggest that the excessive oxygen free radicals resulting from the depression of SOD activity is an important determinant in the pathogenesis of glycerol-induced nephrotoxicity and higher urinary NAG activity is an index of renal tubular cell damage in the glycerol-induced nephrotoxicity.

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사람의 O-linked-N-acetyl-${\beta}$-D-glucosaminidase 유전자를 함유한 대장균의 배양조건과 효소학적 특성 (Culture Conditions of E. coli Harboring Human O-Linked N-Acetyl-${\beta}$-Glucosaminidase Gene and Enzymatic Properties)

  • 강대욱;조용권;서현효
    • 미생물학회지
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    • 제40권2호
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    • pp.147-153
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    • 2004
  • 단백질의 serine이나 threonine의 수산기에 N-acetyl-${\beta}$-D-glucosamine (O-GlcNAc)으로 변형되는 당쇄는 대부분의 진핵세포에서 광범위하게 일어나는 번역 후 수식의 일종으로 여러 세포 내 현상에 관여하고 있다. O-GlcNAc의 변형정도는 O-GlcNAc transfearse (OGT)와 O-linked N-acetyl-${\beta}$-D-glucosaminidase (O-GlcNAcase)에 의해 조절된다. O-GlcNAcase의 효소활성 조절물질을 탐색하기 위한 시험관 내 검정계를 확립하기 위해 재조합 O-GlcNAcase의 생산을 시도하였다. O-GlcNAcase 발현을 최적화하기 위한 배양조건을 검토한 결과 유도 배양온도 $30^{\circ}C$, 유도제인 L-arabinose 0.02%, 유도 배양시간 5시간 등으로 나타났다. 위의 조건에서 대장균 형질전환체를 배양하면서 한 시간 간격으로 배양액을 채취하여 세포를 파괴하고 얻은 효소용액의 활성은 배양 후 3시간에서 5시간까지는 급격히 증가하였으나 그 이후는 거의 증가하지 않았다. Western blot으로 발현된 단백질 양을 조사한 결과는 활성 그래프와 비슷한 양상을 보였다. 이렇게 생산한 O-GlcNAcase의 최적 반응조건은 pH 6.5, 반응온도 $45^{\circ}C$, 기질의 농도 2 mM, pH 6.5로 나타났다.

Effects of the Administration of p-{N ,N-Bis(2-chloroethyl)amino}-4-phenyl acetyl-amino-2,6-piperidinedione (ck-15) on Rat Kidney

  • Park, Sun-Hee;Choi, Bo-Kil;Lim, Dong-Koo;,
    • Toxicological Research
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    • 제14권3호
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    • pp.365-370
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    • 1998
  • To evaluate the renal toxicity of the antitumor agent, p-{N,N,-Bis(2-chloroethyl)amino}-4-phenyl acetyl-amino-2,6-piperidinedione(CK-15), rats were treated with CK-15 (acute: 50mg/kg. i.p., single and subacute: 5mg/kg, i.p., daily for 7 days). The changes in the body weight, water consumption, kidney weights and urine volume after and during the treatment were observed. The concentrations of urinary creatinine and portein, the activities of N-acetyl-${\beta}$-D-glucosaminidase (NAG), alanine aminopeptidase (AAP), ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GT) and lactate dehydrogenase (LDH) in 24hr urine were also determined. The body weight, water consumption, and urine volume were decreased after the acute and subacute administration. However the weights of kidney were not changed after the treatments. The excretion of creatinine was significantly decreased 1 day after acute administration but, returned to the control value. In subactute administration, the excretion of creatinine was gradually decreased. However, the protein excretion did not changed in both treatment. Those indicate that CK-15 might decrease the metabolic rate of muscle. THe urinary activities of NAG, AAP, ${\gamma}$-GT, and LDH were significantly affected bythe drug treatment. The urinary activities of NAG, AAP and ${\gamma}$-GT were significantly increased 1 day after the acute administration and then returned to the control value. However, the urinary activities of LDH were not changed in acute treatment. In subacute treatment, although the urinary activities of NAG were not changed, those of AAP and ${\gamma}$-GT were significantly increased 2.3 times at 3 days during the subacute administration. Also the urinary activities of LDH were significantly increased at 7 day after the administration. These results indicate that the high and subacute administration might induce a damage in the kidney cells. Furthermore the present results suggest that the toxic effects of CK-15 might be due to the accumulation of the metabolites.

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