Extraction Of Invertase From Yeast 443x10

EXTRACTION OF INVERTASE FROM YEAST 0.25 g Baker’s 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