ENZYMES

DEFINING TERMS
Enzymes are protein molecules that act as biological catalysts on reactions. Almost all
processes in a biological cell need enzymes to occur at significant rates.

A catalyst is any substance which makes a chemical reaction go faster, without itself being
changed. A catalyst can be used over and over again in a chemical reaction: it does not get used
up.

Substrate is a molecule upon which an enzyme act upon.

Metabolic pathways are series of chemical reactions occurring within a cell. In each pathway, a
principal chemical is modified by chemical reactions. Enzymes catalyze these reactions, and
often require dietary minerals, vitamins, and other cofactors in order to function properly.

Metabolism is a step-by-step modification of the initial molecule to shape it into another

product.

CHARACTERISTICS OF ENZYMES
1. Enzymes are specific in nature
Enzymes have a specialty in that they possess an 'active site'. This active site allows other
molecules to bind to the enzyme, and these molecules are known as the 'substrate', and the shape
of the active site allows them to fit perfectly forming the enzyme-substrate complex. The
substrate then unbinds in the form of its product.

Enzymes are said to be specific in their nature, as the shape of the active site will only allow one
molecule to fit. It may either catalyze the breaking up or joining of molecules, forming one or
more products. When the reaction is complete, these products leave the active site, and as said
before - the enzyme is unchanged and can thus receive another substrate molecule.

2. Enzyme Concentration
Increasing enzyme concentration increases the rate of the reaction so long as there is sufficient
substrate, there is more enzyme-substrate complexes being formed and more product being
formed thus faster rate of reaction. The rate of reaction increases linearly with enzyme
concentration.

3. Substrate Concentration
Increasing the substrate concentration increases the rate of reaction, to a certain point known as
the saturation point, which is where every enzyme active site is occupied and so substrate simply
ends up 'queuing up' and waiting for an active site.

4. Temperature
The speed at which molecules move around freely is determined by the temperature giving them
more or less energy to do so - at high temperatures the molecules, the enzymes and the
substrates, move around fastest, and thus there are the most collisions per second, causing
enzyme-substrate complexes to be formed more frequently.

The bonds in the enzyme molecule begin to shake so violently that they begin to break, which
causes the enzyme to lose its shape, which changes the shape of the active site (so the substrate
will no longer fit) and the enzyme is said to be denatured.

The reason the rate of reaction does not immediately drop to 0 is that the enzyme slowly loses
shape as the temperature increases, and so the substrate fits less well and eventually not at all,
and so the catalysis does not occur. The process of denaturing is usually irreversible.

In humans 40ºc is the optimum temperature for enzyme reactions- the temperature at which an
enzyme catalyzes a reaction at the maximum rate. Our bodies are kept at 37ºc to make sure they
never go above 40ºc as enzymes would start to denature at even the slightest upward variation -
which would be extremely dangerous as nearly all reactions within the body rely on enzymes.

A very low temperature on the other hand lowers enzyme activity since low temperatures
inactivates the enzymes.

5. The effect of pH on enzyme activity

pH is a measure of hydrogen ions in a solution, and these affect enzyme activity since the
hydrogen ions can react with the enzyme and change the enzymes shape, deforming the active
site. Depending on where it is normally located in the body, an enzyme will have pH values at
which its structure is most stable. As conditions deviate from this point, the enzyme’s ability to
help along reactions decreases.

Most enzymes work best near a pH of 7, but some enzymes operate most effectively in a
particularly acidic environment, such as the stomach; a neutral environment impairs their
function. As with temperature, too high or low pH for the enzyme will denature the enzyme.

6. Inhibition
There are two types of enzyme inhibition – (substances that inhibit the enzymes’ function).

a. Competitive
b. Non-competitive

They can be harmful in that they can stop a reaction happening, or helpful in stopping a reaction
from running wild - perhaps the end-product of a chain of reactions will be an enzyme inhibitor
to prevent the reaction continuing indefinitely.

a. Competitive
Competitive enzyme inhibitors are named as such because they compete with the substrate for
the enzyme molecule's active sites. They achieve this by being a similar shape and fitting into the
active site, temporarily blocking substrate from entering. This makes the reaction slower, since
there is less chance that genuine substrate will collide with an enzyme and form product.
Competitive inhibitors effect on the enzyme is always reversible.

b. Non-Competitive
Non-competitive enzymes are ones that do not compete, in that that either bind permanently to
the active site or bind elsewhere, deforming the active site. Eventually, they will destroy all
available enzymes, stopping the reaction short even if there is remaining substrate, since all
active sites are either blocked or deformed. Unless the inhibitor only binds to somewhere else on
the enzyme very briefly, the enzyme will be irreversibly unusable for normal enzyme-substrate
complexes to form.

Example: The antibiotic penicillin acts by permanently filling an enzyme required for bacterial
cell wall synthesis.
CLASSIFICATION OF ENZYMES
Enzymes are generally classified on the basis of the type of reactions that they catalyze. 6 groups
of enzymes can be recognized on this basis. The following table lists the 6 groups of enzymes
along with examples.
THE ROLE OF ENZYMES IN FOOD PROCESSING AND PRESERVATION
Enzymes Used in Food Processing

Other enzymes used in food processing

Enzyme Used in Fruit Juice Manufacturing

Enzymes are, widely used in fruit Juice and brewing industries to achieve specific objectives,
which cannot be otherwise, achieved. In addition, brewing industry employs varied but specific
strains of yeast for fermentation of the brew.

The cloudiness of fruit juices and wines is mainly due to pectin, which exhibit are usually
associated with other plant polymers and even cell debris. The pectin is digested by pectolytic
enzymes mixtures prepared. The four enzymes present in the mixture act synergistically to
accomplish, a task, which cannot be achieved by mechanical means. Treatment of fruit pulp with
pectolytic enzyme mixture gives the following benefits:

a. Elimination of juice/wine cloudiness,

b. reduced solution viscosity,
c. Increased juice yields, e.g., a 15% increase in case of white grapes,
d. Shorter fermentation period in case of wine making.

Cheese Making
The milk is warmed and a mixture of two enzymes (chymosin and pepsin) known as rennet,
which is obtained from the fourth stomach of the milk-fed calf, is added. This coagulates the
milk to form 'curds and whey'. The whey is a cloudy liquid which contains some protein and
sugars (including 'milk sugar', lactose), while the curds are precipitated protein which is pressed
and subsequently packed in various sized containers for maturing.

Many modern cheeses are produced using chymosin from fungi or bacteria, and sold as
vegetarian cheese.

Liquid-Centered sweets and Chocolates

Have you ever wondered how the liquid center is placed into chocolates? A mixture of equal
parts glucose and fructose is very soluble in water, whereas sucrose is less soluble; and an
equivalent amount of sucrose in water forms a paste-like solid. If the enzyme invertase (from
yeast) is injected into the chocolate which contains sucrose paste, the sucrose is converted to
glucose and fructose.

High Fructose Corn Syrup

Most of us are aware that, traditionally, sugar was obtained from sugar cane or sugar beet.
However, most of the sugar sweeteners used in processed food nowadays, such as jams, ketchups
and soft drinks, comes from sweetcorn. It is also in many so-called health foods.

The process for making the sweetener, known as 'high fructose corn syrup' High-fructose corn
syrup (HFCS) is produced by processing corn starch to yield glucose, and then processing the
glucose to produce a high percentage of fructose. The process is somewhat complicated,
involving the different enzymes amylase and isomerases.