Optimal Prototypic Temperatures for Hydrolyzing Starch Performed by Human and Fungal Amylase

Abstract

Enzymes are constitutive to accelerating activation energies in cells

allowing the facilitation of bond fractures and or formations. To ascertain

how enzymes are utilized in conjunction with a polysaccharide substrate

such as starch, we monitored the progression of starch hydrolysis by agency

of human and fungal amylase over time under various temperature conditions.

The enzyme amylase was introduced to the starch polymer after both had

habituated for 5 minutes in their allotted gelid-to-incalescent water baths

to then be dispensed into spot plates primed with the indicator Iodine in

2-minute intervals up to 10 minutes to surmise qualitatively how much, if

any, starch hydrolysis eventuated. Catalysis increased in both of the

amylase (fungal and human) and starch solutions under their respective

optimal prototypic temperatures, which should be reflective of that

enzyme's natural environment.

Introduction

Enzymes are integral in altering a cell's activation energy in order to

actualize chemical reactions (Raven et al., 2011). Enzymes such as amylase,

which in humans, is produced in the salivary glands and pancreas have a

considerable ability to metabolize long strands of complex polymer units

into simple monomer units (Meisler and Ting, 1993; Simms et al., 2010). For

this reason, they are pivotal in glycogen and dietary starch digestion

(Meisler and Ting, 1993). Without enzymatic collaboration, metabolic

functions would be severely hindered.

Enzymes are most commonly encountered in a globular protein form and

associate with molecules--the substrates--that are subjected to the reaction

(Simms et al., 2010; Raven et al., 2011). The disposition of substrates to

hitch to invaginations on specialized loci--active sites on an

enzyme--engenders an enzyme-substrate complex (Raven et al., 2011). This

complex begets two distinct processes that allow starch to hydrolyze to its

glucose monomer unit by liquefaction and saccharification, with the former

emulsifying it first into an oligosaccharide and the latter further

breaking it down into a monosaccharide (Park et al., 2005; van der Maarel

et al., 2002).

The indicator, Iodine, is used to observe whether any catalytic reactions

such as hydrolysis are taking place as it has an affinity to the

polysaccharide by turning it blue/black in its presence and yellow in

absentia (Simms et al., 2010). The enzyme amylase will be introduced to the

starch to initiate this process. After the requisite time intervals,

qualitative measurements will be taken by visually discerning the

effectiveness of the amylases in their relegated temperatures by consulting

a color-coding schematic. The starch's polysaccharide units should

incrementally start to break down until it is no longer present, providing

that the enzyme is able to function effectively under the temperature

condition it is being subjected to. In this manner, we can surmise the

ranges of optimal temperature, which is just one aspect of optimal

conditions of which PH and temperature are key (Alva et al., 2007;

Copeland, 2000).

Methods

Initial preparation. A mise en place of 2 spot plates placed adjacently to

one another and atop napkins so that above the horizontal row of four

shallow wells the Temperatures (0o, 40o, 60o, and 95o Celsius), can be

written from left to right. On the napkin down the left vertical column of

six shallow wells the Time (0, 2, 4, 6, 8, and 10 min), can be written from

top to bottom. In addition, 4 test tubes were each labeled with a

corresponding temperature as listed above and their enzyme source (H for

Human and F for Fungal). Repeat the labeling process with an additional 4

test tubes, but write the letter S (for starch solution) to differentiate

them from those that will contain the amylase enzyme. To those S marked

tubes add 5mL of 1% starch solution. To the original tubes reserved for the

amylase, add the requisite amount of Human amylase (H), which was obtained

by harvesting 5mL of a group member's saliva into a weighing boat and

transferring 0.5mL of said sputum to each test tube and diluting it with

0.5mL of distilled water.

Timed procedure. Place all 8 test tubes (those containing only starch

solution and those containing only amylase) into their relevant

temperatures (ice bath or water baths). It should be noted that this is a

four-person job as all of the following procedures are performed

simultaneously.