蔗糖磷酸化酶 (Spase)

CAS: 9074-06-0 货号: rp221981 EC号: 851-947-9 PubChem CID: 160315322
有货
级别和纯度: 生物活性 ? 生物活性级(Bioactive)—— 经验证在功能检测中保持生物活性。当分子须具有功能活性而不仅是纯度时使用。 重组 ? 重组(Recombinant)—— 通过重组表达生产,序列确定、批次稳定。适用于来源明确、无动物源的可重现蛋白。 ActiBioPure™ ? ActiBioPure™ —— 阿拉丁面向生物活性和重组产品的优质系列。当同时需要高纯度和保持生物活性时使用。 EnzymoPure™ ? EnzymoPure™ —— 阿拉丁的高质量酶解决方案系列。当酶纯度和明确活性决定检测或工艺性能时使用。 expressed in E.coli,≥50 U/mg powder
Accession #
Q59495
表达系统
E. coli
生物活性
≥50 U/mg powder
★
规格
库存
价格
数量
100U
rp221981-100U
现货 Stock Image
¥1,399.90
250U
rp221981-250U
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¥2,499.90
1KU
rp221981-1KU
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¥7,599.90
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为什么选择此级别

ActiBioPure™,EnzymoPure™,生物活性,重组 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。

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储存与运输

-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。

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质量文档

SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。

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文献证明

在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。

概述


The enzyme is useful for the determination of inorganic phosphate in clinical analysis. 

Reaction: 
Sucrose + Orthophosphate →D-Fructose + α-D-Glucose 1-phosphate

Properties
Molecular weight: ca. 56 kDa (gel filtration) 
Structure: monomer of 56 kDa (SDS-PAGE) 
Isoelectric point: 4.6 
Michaelis constant: 3.9×10^-2M (sucrose); 6.2×10^-3M (phosphate) 
pH Optimum: 7.5           Fig.1
pH Stability: 5.0–8.0           Fig.2
Optimum temperature :40℃           Fig.3
Thermal stability below: 45℃           Fig.4
Stability (liquid form): stable at 37℃ for at least two weeks           Fig.5
Stability (powder form): stable at 30℃ for at least two weeks           Fig.6
Inhibitors: glucose, glucono-1,5-lactone 
Specificity: sucrose (100), maltose (0), starch (0) 
Stabilizer: Sucrose 
Assay Procedure
Principle

 

The appearance of NADPH is measured spectrophotometrically at 340 nm.
Reagents
A. ‌Triethanolamine (TEA) buffer, 0.1 M; pH 7.6‌: Dissolve 1.86 g of triethanolamine hydrochloride in 90 ml of distilled water, adjust to pH 7.6 with 5 N NaOH, and dilute with distilled water to 100 ml.
B. ‌Potassium phosphate buffer‌, (a) ‌0.1 M; pH 6.8‌, (b) ‌0.05 M; pH 6.8‌: (a) Mix 0.1 M KH₂PO₄ and 0.1 M K₂HPO₄ to make a pH 6.8 solution. (b) ‌Dilute 0.1 M potassium phosphate buffer (Reagent B (a)) with the same volume of distilled water.
C. ‌Sucrose solution, 0.32 M‌: 11.0 g of sucrose/100 ml of distilled water.
D. ‌EDTA solution, 9.9 mM‌: 37 mg of EDTA·Na₂·2H₂O/10 ml of potassium phosphate buffer (Reagent B (b)).
E. ‌NADP⁺ solution, 12 mM‌: 9.2 mg of NADP⁺·Na/1.0 ml of distilled water.
F. ‌d-Glucose-1,6-diphosphate (G-1,6-P₂) solution, 0.1 mM‌: 1.0 mg of G-1,6-P₂ cyclohexylammonium salt/10 ml of distilled water.
G. ‌MgCl₂ solution, 1.0 M‌: 4.06 g of MgCl₂·6H₂O/20 ml of distilled water.
H. ‌α-Phosphoglucomutase (α-PGM) suspension, 2000 U/ml‌: Crystalline suspension, 10 mg/ml (200 U/mg).
I. ‌Glucose-6-phosphate dehydrogenase (G6PDH) solution‌: 2000 U/ml in potassium phosphate buffer (Reagent B (b)).
Sample: Dissolve the lyophilized enzyme to a volume activity of 0.8–1.5 U/ml in ice-cold TEA buffer (Reagent A) immediately before measurement.
‌Preparation
1. Pipette the following reagents into a cuvette (light path: 1cm).

1.5 ml   Potassium phosphate bufer   (Reagent B (a))
1.5 ml   Sucrose solution   (Reagent C)
0.03 ml   EDTA solution   (Reagent D)
0.1 ml   NADP⁺ solution   (Reagent E)
0.1 ml   G-1,6-P₂ solution   (Reagent F)
0.05 ml   MgCl₂ solution   (Reagent G)
0.01 ml   α-PGM suspension   (Reagent H)
0.01 ml   G6PDH solution   (Reagent I )
2. Equilibrate at 25°C for about 5 min.
3. Add ‌0.02 ml‌ of sample and mix.
4. Record the increase of absorbance at ‌340 nm‌ in a spectrophotometer thermostated at ‌25°C‌, and calculate the ‌ΔA per min‌ using the linear portion of the curve (denoted as ‌ΔAs‌).
The blank solution is prepared by adding ‌TEA buffer (Reagent A)‌ instead of sampleΔA0‌.
Calculation
Activity can be calculated by using the following formula:


6.2‌: Millimolar extinction coefficient of NADPH at 340 nm‌ (cm²/μmol).
df‌: ‌Dilution factor‌:
‌C‌: Content of sucrose phosphorylase preparation in sample‌ (mg/ml).

EXPERIMENTAL DATA




规格

产品名称
蔗糖磷酸化酶 (Spase)
别名
蔗糖转葡糖基酶
英文别名
sucrose phosphorylase | disaccharide glucosyltransferase | sucrose glucosyltransferase | SP | Sucrose : orthophosphate α-D-glucosyltransferase | EC 2.4.1.7
规格或纯度
生物活性, 重组, ActiBioPure™, EnzymoPure™, expressed in E.coli,≥50 U/mg powder
生物活性
≥50 U/mg powder
表达系统
E. coli
预测分子量
56 kDa
CAS编号和信息
9074-06-0
酶学委员会编号
2.4.1.7
分子类型
酶
产品线
生物产品
储存与运输
物理形态
冻干(Lyophilized)
浓度
expressed in E.coli,≥50 U/mg powder
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
稳定性与储存
长期储存-20℃(2年);避免反复冻融。
单位定义
One unit (U) is defined as the amount of enzyme which produces 1 μmol of NADPH per min at 25°C and pH 6.8 under the conditions described below.

技术文档

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高级数据

三维结构
交互式化学结构模型





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批号(Lot Number) 证书类型 货号
ZJ25F0926780 分析证书 rp221981
ZJ25F0926779 分析证书 rp221981
ZJ25F0926778 分析证书 rp221981
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