• Title/Summary/Keyword: $\beta$-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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    • 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.

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 (Testosterone과 dibutyryl cyclic AMP가 거세한 흰쥐 부정소의 $\beta$ -glucosidase와 몇가지 glycosidase 활성에 미치는 영향 및 부정소 상피세포의 여러 유형에 관한 연구)

  • 최임순;정경순
    • The Korean Journal of Zoology
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    • v.32 no.3
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    • pp.290-303
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    • 1989
  • The activities of $\beta$-glucosidase, $\beta$-glucuronidase and N-acetyl-$\beta$-glucosaminidase were measured to investigate the relationships of them to sexual maturity. Peritoneal injections of testosterone and dibutyryl cAMP to rats were carried out. As a result, the activities of $\beta$-glucosidase and N-acetyl-$\beta$-glucosaminidase were significantly decreased from the third day and that of P -glucurondiase on the seventh day in the castrated groups. In addition, ihe activities of these three enzymes were significantly increased in the testosterone treated groups for 7 days. In case of dbcAMP injection, the activities of these three enzymes were similar to those of castrated groups or had a tendency to be decreased. On electron microscopic examination, principal cells, basal cells and narrow cells were observed in all regions of epididymis. Principal cells were general forms of columnar epithelial cells. Narrow cells had a number of small vesicles and light cells showed low electron density in comparison to other epithelial cells in cauda epididymis. Halo cells were migrating leucocytes btween epithelial cells.

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

  • Ji, Geun-Eog;Lee, Se-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.25 no.2
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    • pp.191-195
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    • 1993
  • The intestinal microflora of humans is an extraordinarily complex mixture of microorganisms, the majority of which are anaerobic bacteria. Amongst them, most prevalent bacteria are Bacteroides, Eubacterium, Peptococcus, Bifidobacteria. We isolated a Bacteroides fragilis strain from a Korean adult and examined various glycosidase activities of this strain. The activities of $N-acetyl-{\beta}-glucosaminidase,\;{\alpha}-fucosidase$, ${\beta}-glucuronidase$, chitobiase and PNPCase were stronger in Bacteroides fragilis Roid 8 than in other intestinal anaerobic bacteria. $N-acetyl-{\beta}-glucosaminidase$ was strongest, followed by ${\alpha}-fucosidase$, ${\beta}-glucuronidase$ and PNPCase. The activities of ${\beta}-galactosidase$, ${\beta}-xylosidase,\;{\alpha}-arabinofuranosidase$ were not present or very low. The activities of ${\alpha}-glucosidase$, ${\beta}-glucosidase$ and ${\alpha}-galactosidase$ were present but at a lower level than in Bifidobacterium. The effect of the carbon sources on the production of $N-acetyl-{\beta}-glucosaminidase$, ${\alpha}-fucosidase$, ${\beta}-glucuronidase$ and PNPCase of Bacteroides fragilis Roid 8 was investigated. :.actose and glucose lowered the production of the varous glycosidase enzymes studied in this work. In addition, we investigated the optimum temperature and pH of each glycosidase from Bacteroides fragilis Roid-8 using crude enzyme preparations.

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

  • Kang Byong-Kyu;Yang Gun-Muug
    • Journal of Veterinary Clinics
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    • v.5 no.1
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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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A Study on Urinary N-acetyl-$\beta$-D-glucosaminidase Activities of Office Workers in a Certain Industrial Complex Area (모 공단지역 사무직 근로자들의 요중 N-acetyl-$\beta$-D-glucosaminidase 역가에 관한 연구)

  • Kim, Hwa-Sung;Lee, Gap-Soo;Lee, Sung-Soo;Ahn, Kyu-Dong;Lee, Byung-Kook
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.3 s.47
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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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    • v.55 no.4
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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.

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 (Glycerol이 흰쥐 신장에서의 Malondialdehyde 함량과 Superoxide Dismutase 활성도 및 요중 단백질 배설량과 $N-acetyl-{\beta}-D-glucosaminidase$ 활성도에 미치는 영향)

  • Shin, In-Chul;Koh, Hyun-Chul
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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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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Culture Conditions of E. coli Harboring Human O-Linked N-Acetyl-${\beta}$-Glucosaminidase Gene and Enzymatic Properties (사람의 O-linked-N-acetyl-${\beta}$-D-glucosaminidase 유전자를 함유한 대장균의 배양조건과 효소학적 특성)

  • 강대욱;조용권;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.147-153
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    • 2004
  • Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.

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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    • v.14 no.3
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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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