c e n z enzymes h y cholesterol e m dehydrogenase se
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O R I G I N Nocardia sp.
� S P E C I F I C A T I O N SAppearance Freeze dried powderActivity ≥5 U/mg powder
� A P P L I C A T I O NUsed in the formulation of Cholesterol testing reagents or in biosensor applications.
� U N I T D E F I N I T I O NOne unit of activity is defined as the amount of enzyme that will catalyse the productionof 1.0 micromole of NADH per minute at 25°C under standard assay method conditions.
CholesterolDehydrogenase
E N Z Y M E S
CAT# C
HDE-70-1241
C A T # C H D E - 7 0 - 1 2 4 1
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� A S S AY P R I N C I P L ECholesterol Dehydrogenase (CDH) catalyses the following reaction:
Cholesterol + NAD+ Cholest-4-en-3-one + NADH + H+
The appearance of NADH is measured at 340nm by spectrophotometry.
CDH
Molecular Weight: 37kDa (SDS-PAGE)
Isoelectric Point: 4.5
Kmvalues: Cholesterol; 1.5 x 10-4 MNAD+; 2.3 x 10-4 M
Optimum pH (Fig. 1): pH 10.0
Optimum Temperature (Fig. 2): 30°C
pH Stability (Fig. 3): 6.5 to 7.5 (37°C for 15 minutes)
Thermal Stability (Fig. 4): Stable at 35°C and below (pH 7.0 for 5 minutes)
Inhibitor: Ag+
Activator: Triton X-100
� C H A R A C T E R I S T I C S
CAT# C
HDE-70-1241
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ME
SRELATIVE ACTIVITY (%)
pH
120
100
80
60
40
20
0
7 8 9 10 11
F IGURE 1 : OP T IMUM pH
F IGURE 4 : THERMAL S TAB I L I TY(pH 7 . 0 FOR 15 M INS . )
F IGURE 2 : OP T IMUM T EMPERATURE
F IGURE 3 : pH S TAB I L I TY ( 37 °C FOR 15 M INS . )
pH
6 7 8 9 10 11
RELATIVE ACTIVITY (%)
TEMPERATURE (°C)
TEMPERATURE (°C)
120
100
80
60
40
20
0
20 30 40 50 60
RESIDUAL ACTIVITY (%)
120
100
80
60
40
20
0
RESIDUAL ACTIVITY (%)
120
100
80
60
40
20
0
10 20 30 40 50 60 70
SUBSTRATE RELATIVE ACTIVITY (%)
Cholesterol 100%
β-Sitosterol 52%
Ergosterol 50%
Stigmasterol 30%
Pregnenolone 14%
Lanosterol 0%
SUBSTRATE RELATIVE ACTIVITY (%)
Dehydroisoandrosterone 0%
Cholic acid 0%
Testosterone 0%
NAD 100%
NADP 0%
TABLE 1 : SUBS TRATE SPEC I F IC I TY OF CHOLES T EROL DEHYDROGENASE
CHEMICAL CONCENTRATION RELATIVE ACTIVITY(mM) (%)
None — 100%
MnCl2 1.0 52%
MgCl2 1.0 50%
CaCl2 1.0 14%
FeCl2 1.0 0%
CHEMICAL CONCENTRATION RELATIVE ACTIVITY(mM) (%)
LiCl2 1.0 96%
NiCl2 1.0 96%
ZnCl2 1.0 48%
AgNO3 1.0 0%
TABLE 2 : E F F ECT OF VAR IOUS CHEMICALS ON CHOLES T EROL DEHYDROGENASE
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CAT# C
HDE-70-1241
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