β-Hydroxybutyrate dehydrogenase (β-HBDH), CAS No.9028-38-0

CAS: 9028-38-0 Cat. No.: rp216193 Molecular Weight: 27 kDa EC Number: 232-830-6

GRADE & PURITY Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™, ≥90%(SDS-PAGE), ≥300U/mg enzyme powder

Accession #

Q02338

Expression system

E. coli

Bioactivity

≥300U/mg enzyme powder

Overview

Application
It is used for the development and mass formulation of β-hydroxybutyrate reagents.
Enzymatic properties

Source: Microorganism
Enzymology Committee Number: EC1.1.1.30
Molecular weight: 27 kDa (SDS-PAGE)
Isoelectric point: 6.1
Km value: 1.46×10-2M (D-3-Hydroxybutyrate), 9.5×10-5M (NAD+)
Inhibitor: Hg²⁺，Ag⁺，SDS 
Optimal pH: 8.5                                                                                                 Figure 1
Optimum temperature: 60℃                                                                             Figure 2
pH stability: 5.0-10.0 (25℃,16h)                                                                       Figure 3
Thermal stability: Stable below 50℃ (pH8.5, 30min)                                        Figure 4
Stability: -25 ~ -15℃ standing store for 12 months. More than 90% activity    Figure 5

 

Assay method for activity
1. Principle

The amount of NADH produced by the reaction can be measured by spectrophotometer at 340nm.
2. Definition of enzyme activity
Unit enzyme activity is defined as the amount of enzyme required to produce 1μmol of NADH per minute under the following reaction conditions.
3. Reagent preparation
Reagent I: 100mM pH8.5 Tris-HCl buffer.
Reagent II: 158mM sodium 3-hydroxybutyrate solution (dissolved with reagent I).
Reagent III: 27.9mM NAD+ (dissolved with reagent I).
Reagent IV: 100mM Tris-HCl, pH8.5, containing 0.1%BSA.
Sample to be tested: Dilute the enzyme solution with reagent IV to 0.1-0.5 U/mL.
4. Operation procedure
4.1 Add 3mL reagent I, 0.5mL reagent II and 0.2mL reagent III into a 3mL colorimetric dish and preheat it at 37°C for 5min.
4.2 Add 100μL sample to be tested and mix well.
4.3 Reaction at 340nm at 37°C, record absorbance change within 1min (∆As)
* Replace enzyme liquid with enzyme diluent, other steps are the same, the absorbance of the resulting solution is blank absorbance (∆Ab)
∆A=∆As-∆Ab

5. Vitality computing
The formula can be captured or edited in this position
Vt: total volume of reaction liquid (3.1mL);
Vs: enzyme liquid volume (0.1mL);
1.0: optical path length (cm);
df: dilution ratio;
C: Enzyme concentration (mg/mL);
6.22: Under standard reaction conditions, the millimolar absorption coefficient (cm2/μmol) of the color-producing group at 340nm.

Specifications

Product Name

β-Hydroxybutyrate dehydrogenase (β-HBDH), CAS No.9028-38-0

Grade

ActiBioPure™, Bioactive, High Performance, Recombinant, EnzymoPure™

Specifications & Purity

Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™, ≥90%(SDS-PAGE), ≥300U/mg enzyme powder

Bioactivity

≥300U/mg enzyme powder

Accession #

Q02338

Predicted molecular weight

27 kDa (SDS-PAGE)

CAS

9028-38-0

Molecule Type

Enzyme

Storage and Shipping

Concentration

≥300U/mg enzyme powder

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20℃ long term (24 months). Upon reconstitution, it is recommended to aliquot. Avoid freeze/thaw cycle.

Unit definition

The amount of enzyme required to produce 1 μmol of NADH per minute under the following reaction conditions.