Final%20Lab - Enzyme Activity Lab Partners Vee...

Enzyme Activity Lab
Partners: Vee Methavarayuth, Katherine Ky, Rowena Gasmen,
Shabana Shah & Carmela Fister
Jennifer Tai
G00398931
Instructor: Vincent Hermoso
June 17, 2008

Introduction:
Enzymes are organic catalysts that are able to speed up chemical reactions
without being consumed by the reactions. The presence of enzymes is the reason why
many chemical transformations do not take place at an extremely slow rate. They are
entities that are reusable. "A catalyst is a chemical agent that speeds up a reaction without
being consumed by the reaction; an enzyme is a catalytic protein (Campbell and Reece).”
Most of these organic catalysts are proteins that posses a specific and unique three-
dimensional structure that is critical to its function. “Enzymes possess active sites where
the actual chemical transformations occur. Substrates are chemicals that undergo
chemical change and which fit into the active site (Fox and Christensen).” The enzymes
and substrates are all very specific in each functions and reactions. With the enzymes not
being consumed within a reaction, it allows the enzyme to "be reused repeatedly on the
same small molecules to build a long chain of repeating subunits" (J.R. Wanamaker). The
enzyme that we used in our experiment is amylase. Amylase is one of the predominant
enzymes found in germinating seeds. It catalyzes the degradation of starch to maltose
which is then subsequently degraded by maltase to glucose. These experiments that we
performed are used for the understanding of the relation between the enzyme of Amylase
and its substrate concentrations in terms of producing a product. In this particular
experiment – we used Porcine Amylose – and we also want to understand how the pH of
the reaction system can alter the efficiency of the Porcine Amylose, and to understand
how the temperature of the reaction system can alter the efficiency of Porcine Amylose.
Hypothesis:
Experiment B : We hypothesized that as time increases, all the substrates will
eventually decrease, following the manual's test that "as absorbance decreases, this
indicates that starch concentration is decreased."
Experiment C: We hypothesized that as time increases, the absorbance will also
decrease from it’s initial measurement; because as time increases, the activity of the
porcine amylase under various pH levels will decrease.
Experiment D: We hypothesized that in various temperatures – as time increases,
absorbance will also decrease, following the manual’s test that “as Absorbance decreases,
this indicates that starch concentration is decreasing”.

Materials:
The materials that we used in each of the experiments are:
Experiment B : Flasks, pipettes, vortexer, spectrophotometer, cuvette, Lugol’s
iodine, starch solution, water. First, we measured 28 mls of enzyme, and diluted the