• Bulletin of the Korean Chemical Society
• /
• 제4권3호
• /
• pp.139-143
• /
• 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.

• 한국임상수의학회지
• /
• 제5권1호
• /
• pp.23-30
• /
• 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.

• Journal of Preventive Medicine and Public Health
• /
• 제27권3호
• /
• pp.547-556
• /
• 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.

• Clinical and Experimental Pediatrics
• /
• 제55권4호
• /
• pp.136-142
• /
• 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.

• 한국축산식품학회:학술대회논문집
• /
• 한국축산식품학회 2002년도 정기총회 및 제29차 춘계국제 학술발표대회
• /
• pp.169-169
• /
• 2002

• 대한약리학회지
• /
• 제32권2호
• /
• pp.259-267
• /
• 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.

• 미생물학회지
• /
• 제40권2호
• /
• pp.147-153
• /
• 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.

• 미생물학회지
• /
• 제40권2호
• /
• pp.87-93
• /
• 2004

세포질과 핵단백질의 serine과 threonine 잔기에 O-linked N-acetyl-$\beta$-glucosamine (O-GlcNAc)의 첨가는고등 진핵 세포에서 흔히 일어나는 번역 후 단백질의 변형 중 하나로서 단백질의 인산화와 유사한 세포 내 신호전달에 관여하는 것으로 보인다. O-GlcNAc의 첨가와 제거는 O-GlcNAc transferase (OGT)와 O-linked N-acetyl-$\beta$-D-glucos-aminidase (O-GlcNAcase) 효소에 의해 각각 촉매된다. 두가지 종류의 사람 유래 O-GlcNAcase 유전자(O-GlcNAcase, v-O-GlcNAcase)를cloning하고 세 가지의 융합단백질로 대장균에서 생산을 시도하였다. O-GlcNAcase의 기질 유사체 인 ${\rho}$-nitrophenyl-N-acetyl-$\beta$-D-g1ucosaminide (${\rho}$NP-$\beta$-D-GlcNAc)를 기질로 사용하여 효소활성을 측정 한 결과 v-O-GlcNAcase는 활성을 나타내지 않았다. 여러 종류의 amino sugar 기질 유사체를 사용하여 O-GlcNAcase의 활성을 측정하였으나 오직 ${\rho}$NP-$\beta$-D-GlcNAc만이 활성을 보였다. Blast검색으로 분석한 결과 아미노 말단의 hyaluronidase-like domain (hyaluronidase-유사 영역)과 카르복시 말단의 N-acetyltransferase 영역 두 곳의 conserved domains 존재하였다. 효소촉매에 중요한 영역을 밝히기 위해 여러 deletion mutants(결손 변이체)를 제작한 후 효소활성을 측정하고 Western blot으로 분석하였다. Hyaluronidas-유사 영역, 유전자 내부와 N-acetyltransferase 영역을 제거할 경우 효소활성이 사라졌으나 아미노 말단의 55개 아미노산과 카르복시 말단의 truncation은 활성을 일부분 유지하였다. 위의 사실에 기초하여 hyaluronidas-유사 영역은 효소활성에 중요하고 카르복시 말단의 N-acetyltransferase 영역은 조절기능으로 작용하는 것으로 추정된다.

• Toxicological Research
• /
• 제14권3호
• /
• pp.365-370
• /
• 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.

• 한국유기농업학회지
• /
• 제25권2호
• /
• pp.311-328
• /
• 2017

두둑과 고랑을 재활용한 한국형 무경운 농업에서 유기물 투입과 관수 효과를 구명하고자 무경운 토양에서 시험을 수행하였다. 1. 토양 미생물상 1회 전량관수 조건에서 대두박 투입 처리구의 토양 세균과 곰팡이 수는 대두박 무 투입구에 비하여 많았다. 그리고 유기질비료 투입량이 표준시비량 66%까지 증가되면 세균과 곰팡이 수는 증가되었으나, 그 이상에서는 세균과 곰팡이 수가 감소되는 경향이었다. 곰팡이/세균 비율은 관수 방법과 관계없이 대두박 투입 처리에서 0.6과 1.1로, 무투입 처리의 0.2와 0.5보다 2배 이상 높았다. 1회 전량 관수 조건에서 유기질 비료 시비량이 증가되면 대두박을 투입한 처리는 방선균 수는 감소되는 경향이었으나, 대두박 무투입에서는 증가되었다. 2회 분할 관수는 1회 전량관수에 비하여 대두박 무 투입 조건에서 세균과 곰팡이 수가 증가되었으나, 대두박 투입조건에서는 방선균 수가 증가되었다. 2. 토양 효소 유기질 비료의 시비량이 증가되면 토양 내 Chitinase 활성은 대두박 투입 토양에서 감소되고, 대두박 무 투입에서는 증가되는 경향이었다. 그러나 대두박을 투입에 관계없이 2회 분할 관수는 1회 전량관수에 비하여 Chitinase 활성이 증가되었다. 1회 전량관수 조건에서 대두박 투입 처리구의 ${\beta}$-Glucosidase 활성은 무투입에 비하여 높았으며, 유기질 비료 투입량이 증가되면 표준시비량의 66%까지는 ${\beta}$-Glucosidase 활성이 증가되었으나, 표준시비량에서는 감소되었다. 대두박 무투입 조건에서 2회 분할관수 토양 내 ${\beta}$-Glucosidase 활성은 1회 전량관수에 비하여 현저하게 증가되었다. 1회 전량관수 조건에서 대두박을 투입한 처리의 N-acetyl-${\beta}$-D-glucosaminidase의 활성은 무투입구에 비하여 높았다. 대두박 투입 처리에서 유기질 비료 투입량이 표준시비량의 66%까지 증가되면 N-acetyl-${\beta}$-D-glucosaminidase의 활성은 증가되었으나, 표준시비량에서는 유의적인 차이가 없었다. 대두박 무투입 조건에서 2회 분할관수는 1회 전량관수에 비하여 N-acetyl-${\beta}$-D-glucosaminidase의 활성은 증가되었다. 대두박 무투입 조건에서 유기질 비료 시비량이 표준량의 66% 수준에서는 토양 내 산성인산가수분해효소(Acid phosphatase)의 활성 높았다. 대두박 투입 조건에서는 유기질 비료 시비량이 증가되면 산성인산가수분해효소(Acid phosphatase)의 활성은 증가되는 경향이었다. 3. 토양 AMF 대두박 무투입 조건에서 유기질 비료의 투입량이 표준시비량의 66%까지 증가되면 토양의 내생균근균의(AMF) 포자수는 증가되었으나, 유기질 비료 투입량이 표준시비량에서는 근균의 포자수는 감소되었다. 그러나 대두박 투입에서 근균의 포자수는 유기질 비료 투입량에 따른 유의적인 차이가 없었다. 그리고 내생 근균의 고추 뿌리에 정착률은 대두박 투입량에 따른 유의적인 차이가 없었으며, 2회 분할 관수도 같은 경향이었다.