Enzymes

 Enzymes are proteins that help speed up metabolism, or

the chemical reactions in our bodies.
 They build some substances and break others down. All living
things have enzymes. Our bodies naturally produce enzymes.
But enzymes are also in manufactured products and food

 Catalytic action
 What is catalytic action of enzyme?

 Enzyme catalysis is the increase in the rate of a process by a biological

molecule, an "enzyme".
 Most enzymes are proteins, and most such processes are chemical reactions.
Within the enzyme, generally catalysis occurs at a localized site, called the active
site.

 Groups of enzymes
There are seven groups of enzymes
1. EC-1: Oxidoreductase

2. EC-2: Transferase

3. EC-3: Hydrolase

4. EC-4: Lyase

5. EC-5: Isomerase

6. EC-6: Ligase

7. EC-7: Translocases
 enzymes with absolute specificity
 Absolute specificity means that one and only one substrate will fit with a particular
enzyme

 enzyme. The enzyme glutamate dehydrogase (1BGV. pdb) will catalyze only the

removal of the nitrogen group from glutamate- NOT any other amino acid. It has absolute
specificity

Absolute specific enzymes will only catalyze one reaction with its specific
substrate.

For example, lactase is an enzyme specific for the degradation of lactose into two sugar
monosaccharides, glucose and galactose.

Condition affecting denaturation

 There are two main causes for enzyme denaturation
 Temperature and PH.

 Enzymes function best at the optimal temperature of an organism. In

the human body, this temperature is 37°C. With slight increases in
temperature, enzyme activity and reaction rates increase, but only to a
certain point.

Food enzymes
 Are enzymes that are safe for consumption and are used by the
food industry during food production to help improve the safety
and quality of foods and the efficiency of the process
 Action of enzymes
 Enzymes are proteins produced by all living organisms.
 They are biological catalysts which are responsible for all chemical
reactions in nature.
 When your body wants to transform food such as starch in bread or
pasta into energy enzymes are used to convert the starch to simple
sugars which can be used by your cells.

Source of enzymes
 Natural Sources of Digestive Enzymes. Fruits, vegetables, and other
foods have natural digestive enzymes.
 Eating them can improve your digestion. Honey, especially the raw
kind, has amylase and protease.

Amylases
Amylase is a digestive enzyme predominantly secreted by
the pancreas and salivary glands and found in other
tissues in very small levels.
 Amylase was first described in the early 1800s and is
considered one of the first enzymes in history to be
scientifically investigated
Types of amylase
 There are three main classes of amylase enzymes; 
 Alpha-, beta- and gamma-amylase, and each act on different
parts of the carbohydrate molecule.
 Alpha-amylase can be found in humans, animals, plants, and
microbes.
 Beta-amylase is found in microbes and plants.
 Gamma-amylase is found in animals and plants.
 What are amylases used for?
Amylases are one of the main enzymes used in industry. Such
enzymes hydrolyze the starch molecules into polymers
composed of glucose units.
Amylases have potential application in a wide number of
industrial processes such as food, fermentation and
pharmaceutical industries.

What are amylases products?

The products of amylase digestion are maltose and
maltotriose (2- and 3-α-1,4–linked molecules,
respectively) and α-dextrins containing 1,6-glycosidic
linkages, because 1,6-glycosidic linkages in starch cannot be
hydrolyzed by amylase.

Cellulases
 Cellulases are enzymes that break down the cellulose found
in plant cell walls into simple sugars that can serve as the
raw materials for biofuels, as well as many of the biobased
chemicals, plastics, and other materials discussed above.

There are three types of cellulases, 

 Endoglucanases, Exocellulases, and Processive
Endoglucanases,
which have different modes of action and different structures.
In common with other enzymes that hydrolyze insoluble
substrates, most cellulases contain a substrate-binding domain
and a catalytic domain (CD).

Glucose isomers
Glucose isomerase (GI) (D-xylose ketol-isomerase; EC. 5.3.
1.5) catalyzes the reversible isomerization of D-glucose
and D-xylose to D-fructose and D-xylulose, respectively.
The enzyme has the largest market in the food industry
because of its application in the production of high-fructose
corn syrup (HFCS).

Glucose invertases
Invertase is an enzyme that catalyzes the hydrolysis
(breakdown) of sucrose (table sugar) into fructose and
glucose. Alternative names for invertase include EC 3.2.

What is invertase used for?

 Invertase is used for the inversion of sucrose in the

preparation of invert sugar and high fructose syrup (HFS).

It is one of the most widely used enzymes in food industry

where fructose is preferred than sucrose especially in the
preparation of jams and candies, because it is sweeter and does
not crystallize easily.
lactase
lactase, also called lactase-phlorizin hydrolase, enzyme found
in the small intestine of mammals that catalyzes the
breakdown of lactose (milk sugar) into the simple sugars
glucose and galactose. In humans, lactase is particularly
abundant during infancy.
 What is pectin enzyme?
 Pectic enzyme is a protein that breaks down pectin in the
fruit. Pectin is the gelatinous material in fruit. It's the stuff that
holds the fruit's fiber together.
 It is also the stuff that causes the resulting fruit juice to have
the appearance of being cloudy. This is known as a pectin haze.

Why is pectinase important?

 Use of pectinases, breaking down the pectin of the fruit and
cell wall degrading enzymes opening more of the plant cells
during fruit juice production, results in significantly higher
yields of enzymes from each ton of fruit processed

Pectin esterase
Pectin esterase (PE):is one of the most heat-resistant enzymes
present in soursop fruit which could lead to gelation and
precipitation of pectin in puree and juice with subsequent loss
of cloud

Polygalacturonase
 Polygalacturonase is a pectinase: an enzyme that degrades
pectin. PGs hydrolyze the O-glycosyl bonds in pectin's
polygalacturonan network, resulting in alpha-1,4-
polygalacturonic residues. The rate of hydrolysis is dependent
on polysaccharide chain length.

Lyase
An enzyme (such as a decarboxylase) that forms double
bonds by removing groups from a substrate other than by
hydrolysis or that adds groups to double bonds.

example of a lyase
 A few examples of lyase include 
phenylalanine ammonia-lyase, citrate lyase, isocitrate lyase, hydroxynitrile,
pectate lyase, argininosuccinate lyase, pyruvate formate lyase, alginate
lyase, and pectin lyase.

protease
 A protease (also called a peptidase or proteinase) is an
enzyme that catalyzes (increases reaction rate or "speeds
up") proteolysis, breaking down proteins into smaller
polypeptides or single amino acids, and spurring the
formation of new protein products.

Types of Protease
 Based on the mechanism of catalysis, proteases are classified
into six distinct classes, aspartic, glutamic, and
metalloproteases, cysteine, serine, and threonine
proteases, although glutamic proteases have not been found in
mammals so far.

serine
Serine is an amino acid. An amino acid is a building block for
protein. Serine comes in two forms: L-serine and D-serine. L-
serine can be consumed in the diet. It can also be made in the
body.
 What is serine used for?
Serine is critical for the production of the body's proteins,
enzymes and muscle tissue.
Serine is needed for the proper metabolism of fats and fatty
acids. It also helps in the production of antibodies.
Serine is used as a natural moisturizing agent in some
cosmetics and skin care products.

Sulfhydryl
 Sulfhydryl SH, a sulfur atom (S) bonded to a hydrogen (H)
atom is a sulfhydryl group. A sulfhydryl compound contains
one or more sulfhydryl groups. Examples include vitamin B-1
and the amino acid cysteine.
A sulfhydryl group (also called “thiol group”) consists of a sulfur
atom with two lone pairs, bonded to hydrogen. The sulfhydryl
group is ubiquitous in our body and mostly found in the
oxidized form as disulfide linkages. The disulfide linkages
contribute to the tertiary and quaternary structures of proteins.

Acid protease
Acid protease A protein-digesting enzyme (see protease) that
exhibits maximum activity and stability in acid conditions (pH
2.0–5.0) and is inactivated at pH values above 6.0. Acid
proteases have a low isoelectric point and are low in basic
amino acids. Two types are widely used in the food and
beverage industries: those from Aspergillus, which resemble
pepsin; and those from Mucor, which resemble rennin
 Peroxidase
Peroxidases are a group of enzymes that catalyze the
oxidation of a substrate by hydrogen peroxide or an
organic peroxide. Most peroxidases are ferric heme proteins;
one notable exception being the glutathione peroxidase, which
is a selenium-containing enzyme. They are present in virtually
all living species.
Peroxidase are enzymes that catalyze oxidation-reduction
reaction by mechanism of free radical that transform
several compounds into oxidized or polymerized products.
The prosthetic group of peroxidase is composed of a protein-
bound heme, usually through a histidine residue that acts as a
proximal ligand.

Catalases
Catalase is a common enzyme found in nearly all living
organisms exposed to oxygen (such as bacteria, plants, and
animals) which catalyzes the decomposition of hydrogen
peroxide to water and oxygen.
It is a very important enzyme in protecting the cell from
oxidative damage by reactive oxygen species (ROS).
Catalase is a key enzyme which uses hydrogen peroxide, a
nonradical ROS, as its substrate.
 This enzyme is responsible for neutralization through
decomposition of hydrogen peroxide, thereby maintaining
an optimum level of the molecule in the cell which is also
essential for cellular signaling processes.
 example of catalase
 Examples include peroxides, superoxide, singlet oxygen,
and hydroxyl radical. In the presence of environmental
stress, they could increase dramatically and lead to oxidative
stress

Glucose oxidase
 Glucose oxidase is a subset of oxidoreductase enzymes that
catalyzes the transfer of electrons from an oxidant to a
reductant.
Glucose oxidases use oxygen as an external electron acceptor
that releases hydrogen peroxide (H2 O2). Glucose oxidase has
many applications in commercial processes, including
improving the color and taste, increasing the persistence of
food materials, removing the glucose from the dried egg, and
eliminating the oxygen from different juices and beverages.
Moreover, glucose oxidase, along with catalase, is used in
glucose testing kits (especially in biosensors) to detect and
measure the presence of glucose in industrial and biological
solutions (e.g., blood and urine specimens).
Hence, glucose oxidase is a valuable enzyme in the industry and
medical diagnostics. Therefore, evaluating the structure and
function of glucose oxidase is crucial for modifying as well as
improving its catalytic properties.
 Finding different sources of glucose oxidase is an effective way
to find the type of enzyme with the desired catalysis. Besides,
the recombinant production of glucose oxidase is the best
 approach to produce sufficient amounts of glucose oxidase for
various uses. Accordingly, the study of various aspects of
glucose oxidase in biotechnology and bioprocessing is crucial.

Lipases
Lipase is an enzyme the body uses to break down fats
in food so they can be absorbed in the intestines.
Lipase is produced in the pancreas, mouth, and stomach.
Lipases (triacylglycerol hydrolases E.C. 3.1.1.3) are enzymes
that catalyze the hydrolysis of triacylglycerols (TAGs) to
glycerol and fatty acids (FAs).
Lipases, together with amylases and proteases, constitute the
three major known digestive enzymes. Plants, animals, and
microorganisms produce lipases.

Examples of these lipases are 

Candida rugosa, Corynebacterium acnes,

Staphylococcus aureus. Additionally, lipases showing fatty
acid specificity is much less common compared to other
groups of lipases.
Geotrichum candidum is the most common fatty acid specific
lipase that shows preferences toward oleic acid.

Phosphatases
A phosphatase is an enzyme that removes a phosphate
group from a protein. Together, these two families of
 enzymes act to modulate the activities of the proteins in a
cell, often in response to external stimuli.

 Based on the optimum pH for the activity, phosphatases

are of two kinds: acid and alkaline. Acid phosphatases
show maximum activity at acidic pH around 6 whereas
alkaline phosphatases show maximum activity at alkaline
pH around 11. The p-nitrophenyl phosphate acts as a
substrate for both acid and alkaline phosphatases.

Lipoxygenase
Lipoxygenases are non-heme iron enzymes that catalyze
the deoxygenation of polyunsaturated fatty acids,
yielding hydroperoxides.
: a crystallizable enzyme that catalyzes the oxidation
primarily of unsaturated fatty acids or unsaturated fats by
oxygen and secondarily of carotenoids to colorless
substances and that occurs especially in soybeans and
cereals