EXTRACTION OF INVERTASE

FROM
YEAST
0.25 g Bakers yeast + d. H2O to make
250 mL

Allow to
Red-brown
+ 3 drops conc. HCl to each tube
(mix
color
well)
Collect supernatant ONLY IF FROTHING
Let it cool

Stand for 20 mins

90oC water bath 5

mins
Denatured enzyme
stock solution
Measure absorbance at 540 nm

Collect supernatant if sedimentation

Enzyme stock

SUCROSE ASSAY
USING
DNS
+ 0.15
mL 0.5
M
CALORIMETERIC METHOD
KOH
Prepare a series of test tubes

Construct a

+ 2.80 mL 0.1 M buffer pH=5 (mix

well)

PREPARATION OF DENATURED INVERTASE

STOCK SOLUTION

100 mL enzyme stock solution

Boiling water bath 10

TUBE
NUMBER

BLANK
1
2
3
4
5
6

ML OF
SUCROSE

ML
DISTILLED

0
+ 3 mL of DNS
0.25
reagent
0.50

1.50
1.25
1.00
0.75
0.75
1.00
0.50
1.25
0.25
95oC water
1.50 bath 10
0
mins

COVER ALL TUBES WITH MARBLES