glycosidic bond. When this bond is cleaved in a hydrolysis reaction, an equimolar mixture of glucose and fructose is produced.

Sucrose + H2 glucose + fructose

Sucrose, a substrate, is considered as the only non-reducing common disaccharide. ! test for sugar detection using ", #dinitrosalicylic acid $ill result in negative readings for sucrose. %ut it can still be applied if sucrose is hydroly&ed first in an acidic solution to produce gluctose and fructose. 'he ob(ectives of the experiment $ere to extract invertase from %a)er*s yeast and to determine the effects of the changes in pH and temperature on reaction rates of an en&yme-cataly&ed reaction. Materials and Methods 'he first procedure done $as the extraction of invertase from the ba)er*s yeast. +.2#g of %a)er*s yeast $as $eighed and dissolved in distilled $ater to ma)e a 2#+ m, solution. 'he solution $as then allo$ed to stand for 2+ minutes at room temperature. 'he supernatant $as collected and transferred into the -rlenmeyer flas) $hich served as the en&yme stoc) solution. 'he follo$ing method $as the preparation of denatured invertase stoc) solution. .++ m, of the en&yme stoc) solution $as incubated in a boiling $ater bath for .+ minutes. 'hen it $as cooled and the supernatant $as collected $hich served as the denatured en&yme stoc) solution. 'able.. /reparation of Standard Solution 'he !ssay using 0initrosalicylic 1olorimetric 2ethod $as performed by first preparing the standard solution using ..#+ m, of .3 glucose solution mixed $ith ..#+ m, of distilled $ater. 'he " m, standard solution $as then mixed $ith " m, 04S reagent. 'he test tubes $ere immersed in 5#1 $ater bath for .+ minutes to develop

the characteristic red-bro$n color. 'he test tubes $ere then cooled to room temperature and the absorbance at #6+ nm $as measured. !fter $hich $as the construction of the hydroly&ed-glucose standard curve done by plotting !#6+ against concentration 7mg8m,9.

:igure..2easurement of the absorbance using Spectrophotometer 'able2. /reparation for the effect of pH on invertase activity

Tube No. pH

1 2

2 "

3 #

4 ;

5 ;.#

6 ;. #

7 <

8 5

Tube No. mL 1% lu!ose standar d solution mL distilled "ater

+."+

+.# +

+.<+

..2+

..#+

..<+

".++

2.;+

2.# +

2.2+

..<+

..#+

..2+

'he effect of pH on invertase activity $as accomplished using t$o test tubes prepared $ith the mixture of . m, buffer solution, +.#+ m, of en&yme stoc) solution, and ..#+ m, of sucrose solution. 'he third test tube $as prepared $ith the mixture of . m, buffer solution, +.#+ m, of denatured en&yme stoc) solution, and ..#+ m, of sucrose solution. !ll test tubes $ere then incubated in =+1 $ater bath for # minutes and $ere added $ith " m, of 04S reagent. 'he three test tubes $ere again immersed in 5#1 $ater bath for .+ minutes to develop