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 Enzymes are proteins that act as catalysts in all living

organisms - microorganisms, plants, animals, and
humans.

 Very small quantities of enzymes can increase the rate

of reactions up to ten million times

 Enzymes operate within a narrow set of conditions,

such as temperature and pH (acidity), and are subject
to inhibition by various means.

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 Enzymes extracted from edible plants and the tissues of food
animals, as well as those produced by microorganisms (bacteria,
yeasts, and fungi), have been used for centuries in food
manufacturing.

 Rennet is an example of a natural enzyme mixture from the

stomach of calves or other domestic animals that has been used in
cheese making for centuries

 Rennet contains a protease enzyme that coagulates milk, causing it

to separate into solids (curds) and liquids (whey).

 For centuries enzymes produced by yeast have been used to

ferment grape juice in order to make wine.

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 • The Egyptians and Sumerians developed fermentation for use in brewing, bread-
2000 BC baking and cheese-making.

• Calves' stomachs and the enzyme, chymosin, were used for cheese-making
800 BC

• The components of yeast cells which cause fermentation were identified and the
1878 term "enzyme" was first used, derived from the Greek term meaning "in yeast".

• Enzymes were first shown to be proteins.

1926

• Enzyme preparations were developed to improve the digestibility and nutrient-

1980s availability of certain animal feeds.

• The first food application of a product of gene technology, alpha-amylase, took

1982 place.

• Recombinant chymosin was approved and introduced in Switzerland, marking an

1988 early approval of a product of gene technology for a food use.

• Two food processing aids obtained using gene technology: an enzyme for use in
1990 cheese-making in the US, and a yeast used in baking in the UK.

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 To date, there have been no reports of consumer allergies to
enzyme residues in food.
 The levels of enzyme residues appearing in foods are so low
that they are highly unlikely ever to cause allergies.
 Like all proteins, enzymes can cause allergic reactions when
people have been sensitised through exposure to large
quantities.
 For this reason, enzyme companies take a variety of
protective measures and some enzymes are produced as
liquids, granules, in capsules or as immobilised
preparations to limit worker exposure.

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 Since the early 1980s, companies which produce enzymes have been
using genetic engineering techniques to improve production
efficiency and quality and to develop new products.

 There are clear advantages here for both industry and consumers,
with major improvements in enzyme production giving better
products and processes.

 However, progress is being slowed down because the debate on

some other, more controversial applications of biotechnology - such
as genetic engineering in animals - is continuing throughout Europe.

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 At present, modern biotechnology can be used to
give a range of advances in enzymatic production
technology:
• Improved productivity and cost-effectiveness in existing
processes. By producing enzymes more efficiently, the amount
of raw materials, energy and water needed to make a product
can be reduced by as much as one-half by changing from a
traditional strain of microbe to a genetically modified one.

• Companies can tailor their enzymes more precisely to

customer demands for products with specific properties.

• Manufacturers can supply enzymes which otherwise could not

be produced in large enough quantities, giving the consumer
access to a wider variety of products. An example is the
amylase-based product which makes bread stay fresh for
longer.

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 ALCOHOL
 BAKING
 BREWING
 STARCH
 CHEESE
 COFFEE & TEA
 DIETICS
 FRUITS & VEGETABLES
 JUICES & WINES
 PROTEINS

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 The production of fermented alcoholic drinks from starch-based raw materials has been
practiced since long
 Raw material: maize (corn), rye, barley and wheat for whisky and other cereals for grain spirits
 starch is still the basic ingredient
 Composed of long chains of glucose molecules and these have to be broken down into smaller
molecules, which the yeast can transform into alcohol
 Enzymes can carry out this process in two stages:
• Liquefaction
• Saccharification
 Traditionally, enzymes have been provided by adding malt
 In many countries malt has been totally replaced from distilling operations by the use of
enzymes
 Advantages of using Enzymes :
• Small quantities of enzyme addition can replace large quantities of malt,
• Resulting easy handling and storage.
• Even raw material costs are reduced by nearly 30% when switching to commercial
enzymes

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 Starch Liquefaction: alpha amylases are used to break down the gelatinised starch to
shorter molecules (dextrins)

 Starch Saccharification: glucoamylase enzyme is used to break down the starch

molecules and the dextrins. This enzyme is capable of achieving the complete
degradation of the starch to fermentable sugars (glucose)

 Fermentation:

• Cereals, particularly maize have low soluble nitrogen.

• This results in poor yeast growth and longer fermentation time.

• By adding a small amount of protease enzyme helps in yeast growth and reduce
fermentation time

• During distillation process it may be necessary to reduce viscosity of the

fermentation broth which is facilitate d by adding beta glucanase / pentosanase
enzymes

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 Bread is the most common and traditional foods around the world
 But bread actually has close links with enzymes. For years, enzymes such
as malt and fungal alpha-amylase have been used in bread making.
 Due to the changes in the baking industry and the ever-increasing
demand for more natural products, enzymes have gained real importance
in bread-making.
 Amylases degrade starch and produce small dextrins for the yeast to act.
 Gluten is a combination of proteins, which form a large network during
dough formation. This network holds the gas in dough proofing and
baking.
 The strength of this network is very important for the quality of all bread
raised by yeast.
 Enzymes such as proteases, xylanases and lipases directly or indirectly
improve the strength of the gluten network and so improve the quality the
bread.

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 A small percentage of pentosans (non-starch polysaccharides) are present in flour.
 Pentosans have an important role in bread quality due to their water absorption
capability and interaction with gluten, which is vital for the formation of the loaf
structure.
 By hydrolysing the pentosans using some enzymes like hemicellulase, pentosanase
or xylanase, the dough becomes easier to handle and the resulting bread has a
bigger loaf volume and an improved crumb structure

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 Alpha amylases have significant effects on baked goods. If the content is low, this leads to low
dextrin production and poor gas production. This in turn results in inferior quality bread
with reduced size and poor crust colour
 To compensate for the deficiencies of the grain, it is necessary to add either sugar or alpha
amylase.
 The addition of enzymes offers certain advantages over sugar. At a flour mill, it is possible to
standardize the enzyme content of the flour so that a uniform commodity can be supplied.
 Enzymes bring about a gradual formation of sugar, which matches the needs of the yeast. When
the dough is placed in the oven, the steadily increasing temperature leads to an increase in the
enzymes' rate of reaction and more sugar is produced.
 Malt flour and malt extract can be used as enzyme supplements as malt is rich in alpha
amylases. However, it is better to use a fungal alpha amylase.
 The alpha-amylases degrade the damaged starch in wheat flour into small dextrins, thus
allowing yeast to work continuously during dough fermentation, proofing and the early stage of
baking. This results in improved bread volume and crumb texture
 In addition, the small oligosaccharides and sugars such as glucose and maltose produced by
these enzymes enhance the reactions for the browning of the crust and baked flavour

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 Requirements of the flour are different from those in bread-making; a
'soft flour' which produces a dough with pronounced plastic properties
is preferred.

 For this purpose, flour with relatively low protein content is desirable

 The gluten protein structure should not be too strong, otherwise the
dough will be too difficult to handle

 Unless flour with these properties is available, it is necessary to add an

agent to weaken the gluten.

 Protein-degrading enzymes soften the gluten without affecting the

other constituents of the dough.

 Several fungal and bacterial proteases can be used for this purpose.
Proteases can also be used when making bread with 'hard flour' i.e.
flour high in gluten protein.
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 In the traditional brewing process where the starch from barley is hydrolysed prior to
fermentation, the barley is allowed to germinate to an extent such that the α and β
amylases are produced for the hydrolytic steps not to be rate limiting

 For Low alcohol beers the mash may be held at 80 ̊ C for a short time denaturing
the heat labile β amyalse and leaving a higher than normal proportion of
unfermentable carbohydrates

 Enzymes that cleave α 1-6 bonds are used to produce low carbohydrate beers
suitable for diabetics

 Protein also plays a vital role in fermentation by providing soluble nitrogen

compounds that the yeast needs

 Since the protein proportion in some types of adjunct is very small, it may be
necessary to provide extra protein in the wort. This can be done by utilising the
protein in the malt more efficiently. By adding a protein splitting (protease) enzyme,
more protein can be solubilised and made available to the yeast.

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 Enzymatic starch conversion, depends on the enzymes used and
syrups with different compositions and physical properties of starch
 There are three basic steps in enzymatic starch conversion:
liquefaction, saccharification and isomerisation.
 Starch liquefaction:
• A starch suspension containing 30-40% dry matter is first gelatinised and
liquefied

• By using heat-stable bacterial alpha amylase, 'maltodextrin' is obtained which

contains mainly different oligosaccharides and dextrins.

• Maltodextrins are only slightly sweet and they usually undergo further
conversion.

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 Starch Saccharification:
• Saccharification is the second step in the process

• Depending of the desired end product, a glucoamylase or a fungal alpha amylase

is used further break down the Maltodextrins

• The glucoamylase can hydrolyse starch completely to glucose along with, a little
maltose and isomaltose

• A pullulanase is a de-branching enzyme that can also be used to aid

saccharification. Fungal alpha amylases can also be added in order to produce
syrups with a higher maltose content, which means high fermentability and a
relatively high degree of sweetness.

 Isomerisation:
• Further going one step ahead, a proportion of the glucose can be isomerised into
fructose, which is about twice as sweet as glucose. An immobilized glucose
isomerase is used

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 Enzymes increase processing capacity and improve economy in the fruit juice and wine
industries
 The most commonly used enzymes in these industries are pectinases which increase juice
yields and accelerate juice clarification.
 They produce clear and stable single-strength juices, juice concentrates and wines, from not
only core-fruits such as apples and pears, but also stone fruits, berries, grapes, citrus-fruits,
tropical fruits and vegetables like carrots, beets and green peppers

 Fruit and Vegetable Juice

 Specialty Enzymes are pectinases that contain hemicellulase enzyme activities
 These products not only increase juice yields, but also increase the color and health-promoting
antioxidants in fruit and vegetable juices extracted by pressing or decanter centrifuge
 By reducing fruit and vegetable mash viscosity and improving solid/liquid separation, they
increase color extraction and juice volume
 Pectinase and Amylase enzyme solutions speed up filtration and prevent storage or post-
packaging haze formation by depectinizing and reducing starch in raw juices. Pectin and starch
must be removed from freshly extracted juices prior to filtration, fining and concentration
 Pectinase and Amylase can reduce starch and pectin in raw fruits and juices, thus achieving
clear and stable juices and juice concentrates
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 Wine

 Many of the biochemical reactions involved in wine production are enzyme-catalyzed

 Winemakers often supplement naturally occurring grape enzymes with commercial enzymes
to increase production capacity of clear and stable wines with enhanced body, flavor and
bouquet
 When added to grapes or musts, Pectinase that contain hemicellulase enzyme products
increase free-run juice volume and extraction of color, fermentable sugars and flavor
components, as well as reduce pressing and fermentation time
 These pectinase or pectinase containing hemicellulase products can increase free-run juice
volume by 20 to 30 percent and lower fermentation time by 30 to 50 percent by reducing
grape-pectin viscosity.
 Rapid clarification and well-separated lees have a positive effect on finished wine flavor,
texture and color
 B-glucanase containing Pectinase depectinize grape-musts during fermentation or young wines
prior to fining and filtration. Grape musts and wines treated with B-glucanase containing
Pectinase are less viscous. They ferment, settle and mature more quickly.
 A beta-glucanase containing pectinase, is also used to degrade Botrytis-glucan. Wines made
from overripe grapes infected with Botrytis cinerea mold are often difficult to clarify and filter
due to high concentrations of Botrytis-produced glucan polysaccharides
 The use of B-glucanase containing Pectinase can speed up clarification and filtration.
 Acid proteases clarify and stabilize some wines by reducing or removing naturally occurring
and yeast synthesized, heat-labile proteins.
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 Proteases are used in both tenderizing and marination of meat

 Papain is the traditional enzyme used

 Others such as:

• bromelain,
• trypsin,
• chymotryosin and
• microbial proteases from Aspergillus are also used

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