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脲酶 来源于刀豆

35篇引用文献 COA COA
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货号 (SKU) 包装规格 是否现货 价格 数量
U128713-250mg
 250mg 现货 Stock Image
U128713-1g
 1g 现货 Stock Image
U128713-5g
 5g 现货 Stock Image
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基本描述

产品名称 脲酶 来源于刀豆
别名 尿素酶
英文别名 Urea amidohydrolase
规格或纯度 EnzymoPure™, ≥45 units/mg dry weight
生化机理 尿素酶催化尿素水解为二氧化碳和氨。尿素酶参与氮代谢和尿素降解。鱼腥草中的尿素酶每个亚基结合 2 个镍离子。
产品介绍


Jespersen (1975) reports that ammonium carbamate is produced in citrate and Tris buffer.
Urease occurs in many bacteria, several species of yeast and a number of higher plants. Varner (1960) has reviewed it. Two of the best sources are: Jack beans (Canavlia ensiformis) from which it has been crystallized and thoroughly studied, and Bacillus pasteurii.
The enzyme is important in assaying for urea. See Guilbault and Montalvo (1970). Its immobilization has been reported: et al. (1974), James and Pring (1975), Messing (1974), Nakamoto et al. (1975), Sundaram (1973) and Tran-Minh and Broun (1975).
Characteristics of Urease from Jack Bean:
Km: 1.3mM in Tris⋅HCl (Cesareo and Langton, 1992).
Specificity
Urease is specific for urea and hydroxyurea (Fishbein and Carbone 1965). See also Sundaram and Laidler (1970).
Composition
Monomeric (a)urease can polymerize to form six unit polymers of about three million daltons. (Fishbein et al. 1970; Fishbein and Nagarajan 1972a). Andrews and Reithel (1970) report on the sulfhydryl groups. Contaxis and Reithel (1971) indicate the molecule can be split in half with no loss of activity. See also Contaxis and Reithel (1972), Fishbein and Nagarajan (1972b), Lynn (1970), and Bailey and Boulter (1969).
Optimal pH：7.4 
Stabilizers
EDTA in concentrations of 1 X 10-3 M. 50% glycerol solutions protect urease crystalline suspension for several months at 4°C.
Inhibitors
Heavy metals. NH4+ ions formed. See also Fishbein and Carbone (1965). Sodium and potassium ions are inhibitors (Cesareo and Langton, 1992).
Urease Assay
Method
Worthington has adopted an assay method where the hydrolysis of urea is measured by coupling ammonia production to a glutamate dehydrogenase reaction.
formulas
One unit results in the oxidation of one micromole of NADH per minute at 25°C and pH 7.6 under the specified conditions. In addition to increased sensitivity, the assay method possesses the advantage that it can be manipulated to permit quantitation of urea.
Reagents
0.1 M Potassium phosphate buffer, pH 7.6
0.023 M Adenosine-5'-diphosphate (ADP) in phosphate buffer
0.0072 M NADH in phosphate buffer
0.026 M a-Ketoglutarate in phosphate buffer
1.8 M Urea in phosphate buffer
Glutamate Dehydrogenase: Dilute to approximately 500 units/ml in 50% glycerol or phosphate buffer. Store cold during use.
Enzyme
Dissolve enzyme at one mg/ml in 0.1 M phosphate buffer, pH 7.6. Immediately prior to use, dilute further in buffer to obtain a rate of 0.02-0.04 ΔA/minute.
Procedure
Adjust spectrophotometer to 340 nm and 25°C. Pipette into each cuvette as follows:

0.10 M Phosphate buffer, pH 7.6 2.4 ml
0.023 M ADP 0.1 ml
0.0072 M NADH 0.1 ml
0.026 M α-Ketoglutarate 0.1 ml
1.8 M Urea 0.1 ml
GLDH (500 units/ml) 0.1 ml

Incubate in spectrophotometer at 25°C for 5-10 minutes to achieve temperature equilibration and establish blank rate, if any. A slight change in absorbance may be observed due to trace ammonia in reagents. Upon obtaining a zero change in absorbance, add 0.1 ml appropriately diluted enzyme. Record decrease in A340 for 8-10 minutes. Determine ΔA340/minute from the linear portion of the curve. A slight lag may occur.

Calculation 

储存与运输

浓度 ≥45 units/mg dry weight
储存温度 -20°C储存
运输条件 超低温冰袋运输
CAS编号和信息 9002-13-5
酶学委员会编号 3.5.1.5
单位定义 One Unit oxidizes one micromole of NADH per minute at 25°C, pH 7.6. The hydrolysis of urea is measured by coupling ammonia production to a glutamate dehydrogenase reaction.

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批号(Lot Number) 证书类型 货号
C2511609 分析证书 U128713
C2511610 分析证书 U128713
C2511619 分析证书 U128713
B2510038 分析证书 U128713
J2409536 分析证书 U128713
J2409537 分析证书 U128713
J2409538 分析证书 U128713
G2317037 分析证书 U128713
G2317079 分析证书 U128713
G2317096 分析证书 U128713
G2426152 分析证书 U128713
H2209752 分析证书 U128713
B2318515 分析证书 U128713

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9. Lingxia Qin, Xinxin Ren, Kaiyue Hu, Di Wu, Zhiyong Guo, Sui Wang, Linwen Jiang, Yufang Hu.  (2022)  Supramolecular Host-Guest Interaction-Driven Electrochemical Recognition for Pyrophosphate and Alkaline Phosphatase Analysis.  CHEMBIOCHEM,  23  (20): (e202200413).  [PMID:35997506] [10.1002/cbic.202200413]
10. Deng Hao-Hua, Yang Hui-Jing, Huang Kai-Yuan, Zheng Yi-Jing, Xu Ying-Ying, Peng Hua-Ping, Liu Yin-Huan, Chen Wei, Hong Guo-Lin.  (2022)  Antenna effect of pyridoxal phosphate on the fluorescence of mitoxantrone-silicon nanoparticles and its application in alkaline phosphatase assay.  ANALYTICAL AND BIOANALYTICAL CHEMISTRY,  414  (17): (4877-4884).  [PMID:35576012] [10.1007/s00216-022-04110-7]
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