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Inactivation Kinetics of β-N-Acetyl-D-Glucosaminidase from Prawn (Penaeus vannamei) by Formaldehyde 被引量:2
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作者 谢晓兰 石艳 +1 位作者 黄乾生 陈清西 《Marine Science Bulletin》 CAS 2006年第1期34-45,共12页
β-N-Acetyl-D-glucosaminidase (NAGase, EC.3.2.1.52) is chitinolytic enzymes and disintegrate dimmer and trimer a composition of oligomers of N-acetyl-β-D-glucosamine (NAG) into monomer. Prawn (P. vannamei) NAG... β-N-Acetyl-D-glucosaminidase (NAGase, EC.3.2.1.52) is chitinolytic enzymes and disintegrate dimmer and trimer a composition of oligomers of N-acetyl-β-D-glucosamine (NAG) into monomer. Prawn (P. vannamei) NAGase is involved in digestion and molting processes. Some pollutants in seawater affect the enzyme activity causing loss of the biological function of the enzyme, which affects the exuviating shell and threatens the survival of the animal. The effect of formaldehyde on prawn (P. vannamei) β-N-acetyl-D-glucosaminidase activity for the hydrolysis of pNP-NAG has been studied. The results show that formaldehyde, at appropriate concentrations, can lead to reversible inactivation of the enzyme, and the IC50 is estimated to be 1.05mol· L^-1. The inactivation mechanism obtained from Lineweaver-Burk plots shows that the inactivation of the enzyme by formaldehyde belongs to the competitive type. The inactivation kinetics of the enzyme by formaldehyde has been studied using the progress-of-substrate-reaction method described by Tsou, and the rate constants have been determined. The results show that k+0 is much larger than k-0, indicating the free enzyme molecule is fragile in the formaldehyde solution. 展开更多
关键词 P. vannamei β-n-acetyl-D-glucosaminidase inactivation kinetics FORMALDEHYDE
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THE RELATIONSHIP BETWEEN THE N-GLYCOSYLATION OF ACETYLGLUCOSAMINYLTRANSFERASE V AND ITS ACTIVITY
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作者 张志兵 郭华北 +3 位作者 王琛 徐丰 陈惠黎 金由辛 《Chinese Medical Sciences Journal》 CAS CSCD 2000年第2期79-82,共4页
Objective. The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V(Glc NAcT-V) and its activity and to know which site among the 6 N-glycosylation sites... Objective. The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V(Glc NAcT-V) and its activity and to know which site among the 6 N-glycosylation sites in the GlcNAcT-V gene is the most important. Methods.Wild type of GlcNAcTV was transfected into COS7 cells and its activity was measured 48 h later. The first site (Asn 110) was mutated with sitedirected mutagenesis and transfected into COS7 cells. Results. It was found that after the cells were added tunicamycin(TM, 1 μ g/ml), the activity was 117% of the wild type. The activity of the cells with mutating GlcNAcTV was about 120% of the wild type RTPCR showed that there was no significant change in mRNA expression among the three groups. Conclusion.The Nglycosylation is important for its activity. Our results suggest that the Nlinked carbohydrates on GlcNAcTV are required for the posttranscriptional activity of the enzyme. 展开更多
关键词 GlcnAcT-V glycosylation site MUTAGEnESIS
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土壤氮素矿化与土壤供氮能力——Ⅳ.土壤有机氮组分及其矿化 被引量:14
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作者 付会芳 李生秀 《西北农林科技大学学报(自然科学版)》 CSCD 1992年第S1期63-67,共5页
利用Bremner法,对盆土不同形态有机氮进行了测定。结果表明,各组分氮与土壤有机质.全氮高度相关,占全氮的百分率大小顺序为:氨基酸N>NH_4^+-N>氨基糖N>非鉴别N。在盆栽和培养两种条件下,各组分氮的变化差异比较明显,对矿化氮... 利用Bremner法,对盆土不同形态有机氮进行了测定。结果表明,各组分氮与土壤有机质.全氮高度相关,占全氮的百分率大小顺序为:氨基酸N>NH_4^+-N>氨基糖N>非鉴别N。在盆栽和培养两种条件下,各组分氮的变化差异比较明显,对矿化氮的贡献大小顺序与占全氮百分率大小顺序相同。 展开更多
关键词 有机氮组分 氨基n 氨基糖n nH4+-n 非鉴别n
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Multiple organ dysfunction syndrome due to ingestion of fish gall bladder 被引量:2
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作者 邓跃林 肖桂林 +6 位作者 金益强 罗学宏 孟新科 李洁 敖翔 肖奇明 周利平 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第7期1020-1022,共3页
OBJECTIVE: To investigate changes in renal function, urine N-acetyl-beta-D-glucosaminidase enzyme (N-AG),liver function, myocardial enzymes, the pathology of renal damage and the mechanism of acute renal failure (ARF)... OBJECTIVE: To investigate changes in renal function, urine N-acetyl-beta-D-glucosaminidase enzyme (N-AG),liver function, myocardial enzymes, the pathology of renal damage and the mechanism of acute renal failure (ARF) associated with fish gall bladder poisoning. METHODS: Eleven patients with acute fish gall bladder poisoning were consecutively admitted to our hospital from September 1997 to October 1999. Renal function, urine N-AG enzyme, liver function, and myocardial enzymes were assayed before and after treatment. One patient consented to a kidney biopsy and the pathology of renal damage was observed under light and electron microscopes. RESULTS: All patients had multiple organ dysfunction syndrome (MODS) and 11 patients suffered from ARF. Ten patients had liver dysfunction, ten patients had poisonous myocarditis, and 8 patients had gastrointestinal dysfunction. Renal function, urine N-AG enzyme, liver function, and myocardial enzymes were significantly improved after treatment compared with those of before treatment (P 展开更多
关键词 FISHES GALLBLADDER ADULT Aged AnIMALS FEMALE Food Poisoning Humans Male Middle Aged Multiple Organ Failure Research Support non-U.S. Gov't
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Endo-H在毕赤酵母中的表达、纯化及其在N-糖基化分析中的应用 被引量:5
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作者 支艳艳 唱韶红 +3 位作者 巩新 宋西勇 吴军 刘波 《军事医学》 CAS CSCD 北大核心 2014年第3期193-197,共5页
目的通过巴斯德毕赤酵母(Pichia pastoris)表达制备内切β-N-乙酰氨基葡糖苷酶-H(endo-beta-N-acetylglucosaminidase H,Endo-H),并用于蛋白质的N-糖基化分析。方法根据GenBank中公布的褶皱链霉菌(Streptomyces plicatus)Endo-H的cDNA序... 目的通过巴斯德毕赤酵母(Pichia pastoris)表达制备内切β-N-乙酰氨基葡糖苷酶-H(endo-beta-N-acetylglucosaminidase H,Endo-H),并用于蛋白质的N-糖基化分析。方法根据GenBank中公布的褶皱链霉菌(Streptomyces plicatus)Endo-H的cDNA序列,设计并合成Endo-H全基因,将其克隆至P.pastoris表达载体pPIC9中,重组质粒转化P.pastoris JC308宿主菌,工程菌经甲醇诱导,分泌表达Endo-H,目的蛋白经疏水层析、凝胶过滤两步纯化后,纯度可>95%。成品用于基于DNA测序的荧光辅助糖电泳(DNA sequencer assisted fluorophore-assisted carbohydrate electrophoresis,DSA-FACE)分析核糖核酸酶B(ribonuclease B,RNaseB)的糖基结构,比较了Endo-H与商品化N-糖酰胺酶F(peptide-N-asparigine amidase F,PNGase F)的糖基切割功能。结果利用P.pastoris表达制备EndoH,可切割天然或变性状态下的β-1,4-糖苷键连接的甘露糖型结构糖链,不能切割复杂型糖链糖蛋白;经DSAFACE分析,结果显示Endo-H酶切RNaseB后其糖链为Man5GlcNAc-Man9GlcNAc,PNGaseF酶切RNaseB的糖链为Man5GlcNAc2-Man9GlcNAc2,两者相差一个GlcNAc。结论通过P.pastoris制备的Endo-H具有天然生物活性,可用于蛋白质的N-糖基化结构分析。 展开更多
关键词 毕赤酵母 n-糖基化
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