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Let’s start with t

this case, the star


addition of acid o
formation of sim
The partial hydro
in which starch g
hot aqueous solu
bonds holding th
Let’s start with the basics. Hydrolysis is simply the breaking down of a compound—in
this case, the starch in grain—through reaction with water and is accomplished by the
addition of acid or enzymes. Total hydrolysis of the starch molecule results in the
formation of simple sugars known as glucose. In ethanol production, this occurs in stages.
The partial hydrolysis of starch known as gelatinization and liquefaction is the condition
in which starch granules are forced to swell and burst (lose their crystalline structure) in a
hot aqueous solution while an enzyme catalyzes the cleaving of some of the hydrogen
bonds holding the glucose polymer (starch) together, producing less viscous dextrin
molecules.
Starch is generally insoluble in water at room temperature. Because of this, starch in
nature is stored in cells as small granules which can be seen under a microscope. Starch
granules are quite resistant to penetration by both water and hydrolytic enzymes due to
the formation of hydrogen bonds within the same molecule and with other neighboring
molecules. However, these inter- and intra-hydrogen bonds can become weak as the
temperature of the suspension is raised. When an aqueous suspension of starch is heated,
the hydrogen bonds weaken, water is absorbed, and the starch granules swell. This
process is commonly called gelatinization because the solution formed has a gelatinous,
highly viscous consistency.

Starch
Polysaccharides are carbohydrate polymers consisting of tens to hundreds to several
thousand monosaccharide units. All of the common polysaccharides contain glucose as
the monosaccharide unit. Polysaccharides are synthesized by plants, animals, and humans
to be stored for food, structural support, or metabolized for energy.

Starch:

Plants store glucose as the polysaccharide starch. The cereal grains (wheat, rice, corn,
oats, barley) as well as tubers such as potatoes are rich in starch.

Starch can be separated into two fractions--amylose and amylopectin. Natural starches
are mixtures of amylose (10-20%) and amylopectin (80-90%).

Amylose forms a colloidal dispersion in hot water whereas amylopectin is completely


insoluble. The structure of amylose consists of long polymer chains of glucose units
connected by an alpha acetal linkage. The graphic on the left shows a very small portion
of an amylose chain. All of the monomer units are alpha -D-glucose, and all the alpha
acetal links connect C # 1 of one glucose to C # 4 of the next glucose.
Acetal Functional Group:

Carbon # 1 is called the anomeric carbon and is the center of an acetal functional group.
A carbon that has two ether oxygens attached is an acetal.

The Alpha position is defined as the ether oxygen being on the opposite side of the ring
as the C # 6. In the chair structure this results in a downward projection. This is the same
definition as the -OH in a hemiacetal.

Starch Coil or Spiral Structure:

As a result of the bond angles in the alpha acetal linkage, amylose actually forms a spiral
much like a coiled spring.

Amylose is responsible for the formation of a deep blue


color in the presence of iodine. The iodine molecule slips inside of the amylose coil.
Amylopectin:

The graphic on the left shows very small portion of an amylopectin-type structure
showing two branch points [drawn closer together than they should be]. The acetal
linkages are alpha connecting C # 1 of one glucose to C # 4 of the next glucose.

The branches are formed by linking C # 1 to a C # 6 through an acetal linkages.


Amylopectin has 12-20 glucose units between the branches. Natural starches are mixtures
of amylose and amylopectin.

In glycogen, the branches occur at intervals of 8-10 glucose units, while in amylopectin
the branches are separated by 10-12 glucose units.
Compare Cellulose and Starch Structures:

Cellulose: Beta glucose is the monomer unit in cellulose. As a result of the bond angles
in the beta acetal linkage, cellulose is mostly a linear chain.

Starch: Alpha glucose is the monomer unit in starch. As a result of the bond angles in the
alpha acetal linkage, starch-amylose actually forms a spiral much like a coiled spring.
Cellulose:

The major component in the rigid cell walls in plants is cellulose. Cellulose is a linear
polysaccharide polymer with many glucose monosaccharide units. The acetal linkage is
beta which makes it different from starch. This peculiar difference in acetal linkages
results in a major difference in digestibility in humans. Humans are unable to digest
cellulose because the appropriate enzymes to breakdown the beta acetal linkages are
lacking. (More on enzyme digestion in a later chapter.) Undigestible cellulose is the fiber
which aids in the smooth working of the intestinal tract.

Animals such as cows, horses, sheep, goats, and termites have symbiotic bacteria in the
intestinal tract. These symbiotic bacteria possess the necessary enzymes to digest
cellulose in the GI tract. They have the required enzymes for the breakdown or hydrolysis
of the cellulose; the animals do not, not even termites, have the correct enzymes. No
vertebrate can digest cellulose directly.

Even though we cannot digest cellulose, we find many uses for it including: Wood for
building; paper products; cotton, linen, and rayon for clothes; nitrocellulose for
explosives; cellulose acetate for films.
Because the bacterial alpha-amylase to be used in this experiment randomly attacks only
the alpha-1,4 bonds, it belongs to the liquefying category. The hydrolysis reaction
catalyzed by this class of enzymes is usually carried out only to the extent that, for
example, the starch is rendered soluble enough to allow easy removal from starch-sized
fabrics in the textile industry. The paper industry also uses liquefying amylases on the
starch used in paper coating where breakage into the smallest glucose subunits is actually
undesirable. (One cannot bind cellulose fibers together with sugar!)
Let’s start with t
this case, the star
addition of acid o
formation of sim
The partial hydro
in which starch g
hot aqueous solu
bonds holding th
It is a protein with catalytic activity. An enzyme is a biological catalyst able to accelerate several folds a
chemical reaction.
All enzymes are proteins, but not all proteins are enzymes.
Enzymes are produced by cells, but enzymes are not capable of reproduction.
Enzymes are, therefore, not "alive", but are biologically active under certain conditions of pH, temperature,
medium composition, etc.
Enzymes are active under relatively mild reaction conditions.
Enzymes are not infectious to individuals or polluting the environment.

Contrary to inorganic catalysts (i.e., heavy metals) enzymes are extremely specific, which means that each
enzyme can break or form a single type of chemical bond in a given chemical compound or structure.
Furthermore, enzyme-catalyzed reactions take place under mild conditions of temperature (30-70°C), pH
(between pH 4.5 and 9), pressure (atmospheric), etc., and are to be viewed as eco-compatible, as they do
not generate polluting residues or toxic by-products.
There are instances where enzymes can be employed to catalyze some reactions even at extreme
temperatures (beyond 100°C, as is the case for high-temperature amylases ) or of pH (up to pH 12, such as
with alkaline proteases used in leather applications).

There are numerous applications of enzymes, in the fields of industrial chemistry and food processing,
amongst which the following should be mentioned:

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