SAINT MARY’S UNIVERSITY

BAYOMBONG, NUEVA VIZCAYA 3700

SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCES
DEPARTMENT
ACTIVITY 9
OPTIMUM PH AND TEMPERATURE OF HYDROLYTIC ENZYMES

Enzyme digestion is necessary for the breakdown of molecules into absorbable forms
and for bioavailability. The enzyme papain is one hydrolytic enzyme that digests many protein
substrates. There are requirements for enzymes to work at an optimum. Examples of these are
pH and temperature.

OBJECTIVES
1. To determine the optimum temperature for papain enzyme to work on proteins
2. To determine the pH requirement of papain enzyme.

MATERIALS
Unripe papaya (diced in small pieces) 0.1 N NaOH
Gelatin powder 0.1 N HCl

PROCEDURES
Slice unripe papaya into small pieces and blend it. Divide these into 5 portions to be
used for the succeeding procedures

I. Effect of temperature
Into 3 small beakers, put blended papaya to make 3 set-ups. Set-up 1 will be at room
temperature, set-up 2 is boiling temperature (cooked papaya) and set-up 3 is 70 ®C heating.
Drain the excess water from all the set-ups. Then measure 3.75 g gelatin and cook in 250 mL
water, stirring constantly. Let it cool but not until it starts to solidify. Pour 50 mL of the cooked
gelatin into each of the three set-ups and wait until it solidifies. Let it stand for 30 more
minutes. Observe which set-up shows more hydrolysis on gelatin. Record your results.

Set-up Observation
1 (Room Temp)
2 (cooked)
3 (70 ®C)

II. Factors the affect enzyme activity

To the observed optimum temperature, make 2 set-ups. To one set-up, add 10 drops 0.1
N NaOH, to the other set-up add 10 drops 0.1 N HCl. Then proceed as the above
procedures. Record your results.
 SAINT MARY’S UNIVERSITY
BAYOMBONG, NUEVA VIZCAYA 3700
SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCES
DEPARTMENT
Set-up Observation
1 (Basic)
2 (Acidic)

QUESTIONS
1. What are other applications of the enzyme papain?
2. Will papain enzyme work as described in commercials for papaya soap? Why or why
not?
GENERALIZATION: