EXTRACTION OF INVERTASE FROM YEAST+ 3 drops conc. HCl to each tube (mix well)0.25 g Bakers yeast + d. H2O to make 250 mL

90oC water bath 5 mins

Stand for 20 mins

+ 0.15 mL 0.5 M KOHCollect supernatant if sedimentation occurs

Enzyme stock solution

+ 2.80 mL 0.1 M buffer pH=5 (mix well)

PREPARATION OF DENATURED INVERTASE STOCK SOLUTION+ 3 mL of DNS reagent

100 mL enzyme stock solution

95oC water bath 10 mins

Boiling water bath 10 mins

Allow to coolRed-brown color

Let it coolCollect supernatant ONLY IF FROTHING OCCURS

Measure absorbance at 540 nmDenatured enzyme stock solution

SUCROSE ASSAY USING DNS CALORIMETERIC METHODConstruct a curve

Prepare a series of test tubes

TUBE NUMBERML OF SUCROSEML DISTILLED

BLANK01.50

10.251.25

20.501.00

30.750.75

41.000.50

51.250.25

61.500

COVER ALL TUBES WITH MARBLES