Enzymes

Definition:
- Protein that acts as biological catalyst.

Biological catalysts:
- Are the substances which speed up a metabolic reaction without being changed or used up.

Substrate:
- Is the substance on which an enzyme acts.
- Each enzyme has a certain substrate on which it acts.
Examples:
1- The enzyme pepsin which is secreted by the walls of the stomach its substrate is protein.

Pepsin
Protein
polypeptides
2- The enzyme amylase which is secreted by salivary glands its substrate is starch.

Starch

Amylase
maltose ( a type of sugar)

The main characteristics of enzymes:

1- are protein in nature
2- are made in cells
3- one enzyme can be used over and over again
4- not changed by the reaction
5- are specific in their function
6- Are destroyed or denatured by temperature and pH

Enzymes are specific: (How

How enzymes act)
-Each
Each enzyme acts on only one kind of substrate.
-Each enzyme has a region in it called the active site into which the substrate fits exactly like a lock and
a key where the enzyme represents the lock and substrate represents the key.
-Each
Each enzyme speeds up a certain reaction only.
- High temperature above optimum change the shape of the active site, therefore
therefore the enzyme cannot act.

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Effect of temperature on enzyme activity:

- The activity of most enzymes increases by the increase in temperature till about 40oC then any
increase in temperature leads to a decrease in the activity of the enzyme.
- Increase in temperature increases rate of reaction as it increases kinetic energy of molecules of both
enzyme and its substrate making them move around faster making it easier for them to react.
- Above 50oC the enzyme denature (means that it loses its shape and activity and cannot regain it even
if the temp. changes). It is irreversible.
Optimum temperature of an enzyme:
- it is the temperature at which the enzyme works best and its most active.

Effect of pH on enzyme activity:

- each enzyme has a certain pH at which it works best.
- Any increase or decrease around this pH decreases
the activity of the enzyme.
- Extremes of pH changes may denature some enzymes.
Optimum pH of an enzyme:
It is the pH value at which an enzyme works best.
Examples:
The optimum pH of amylase is about 7 to 7.5
The optimum pH of pepsin is about 2 to 2.5

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A- Role of enzymes in the germination of seeds:

Seeds contain stored insoluble food with inactive enzymes, when water enters the seed, and in suitable
conditions the enzymes in the seed become active to catalyze the gradual breakdown of stored insoluble
food in the seed to a soluble from that can be used by the embryo till it forms green leaves which are used
making food by photosynthesis.
Examples of such enzymes:
1- amylase: which catalyses the breakdown of stored insoluble starch to a soluble sugar known as
maltose.
2-proteases: which catalyze the breakdown of stored insoluble proteins into soluble amino acids.

B- Role of enzymes in biological washing powder:

-Biological washing powders contain enzymes, such enzymes act as catalysts that help in removing stains
by breaking the insoluble molecules of stains into soluble molecules so they can be washed and removed
by water.
- Stains can be of animal or plant origin e.g. blood, egg yolk, or fruit juice
Examples of the enzymes used in washing powders:
1- Lipases: enzymes which catalyze the breakdown of fats to fatty acids and glycerol, and this is
important in removing greasy stains.
2- Proteases: enzymes which catalyze the breakdown of insoluble proteins into soluble amino acids and
this is important in removing proteins stains such as blood stains.
Advantages of biological washing powder:
1- Save expense of boiling.( these powders work at low temperatures so they save energy)
2- Avoid the damage of clothes by boiling and rubbing.
3- Are not corrosive as strong acids and alkalins.
4- Are not allergic
5- Are biodegradable and wont cause pollution
6- Enzymes are specific, so the process can be controlled.
Note:
1- Enzymes cannot be used for silk, wool or leather, which contain proteins as it may be damaged by the
proteases in the biological washing powder.
2- Hands should be washed after using these powders in order not to be harmed by the enzymes in
biological washing powders.
3- biological powders must be used with warm water to activate enzymes and not boiling water which
denatures enzymes and stops their activity.

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C- Enzymes in food industry:

1- Pectinase enzyme used in juice extraction from fruits.
Pectin is a substance found in plant cell walls which helps to stick plant cells together, when pectinase
enzyme is used, it acts on pectin to be broken down into sugar, this is useful in the following ways:
Makes it much easier to squeeze juice from fruit.
Helps it increasing the volume of the juice produced.
Makes juice clear rather than cloudy.
2-Protease enzymes used in making baby foods
Act on proteins in food to be broken down into small polypeptides or amino acids to be easily absorbed
by digestive system of babies.
3-Amylase used in obtaining sugar from starch
Potatoes, grains or other starch containing substances are crushed and amylase is added to act on starch
for making maltose which is used in sweet making.
4- Isomerase enzyme is used in converting glucose into fructose
As fructose is sweeter so less sugar can be used, this also useful for diabetic.
5- Lactase enzyme is used to break down lactose of milk
Lactase acts on lactose of milk to be digested into glucose and fructose to help those have problems in
digesting lactose of milk.
6- Enzymes and cheese making
An enzyme called rennin extracted from stomach of animals (and now it is produced by genetic
engineering) This enzyme clots milk at the first stage of making cheese.
7- Enzymes are used in brewing and baking:
Yeast contain enzymes that act on sugars to produce alcohol used in beer and wine and also produce
CO2 used in bread.

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THE USE OF MICROORGANISMS IN THE

FOOD INDUSTRIES
Biotechnology: use of microorganisms in industry
Microorganisms: are very small organisms, often unicellular, cannot be seen by the naked eyes.
Examples: bacteria, fungi, yeast and phytoplanktons (microscopic organisms live in water and can carry
out photosynthesis.)

1- Using microorganisms in yogurt making:

1- Homogenization: to form thick milk with uniform distribution of fat (prevent fats from separating) 22- Milk is heated to about 87oC to kill any microorganisms that may be found in milk. (sterilization)
3- Milk is left to cool to about 35-40oC.
(because this temperature is suitable for enzymes of the bacteria which is known as Lactobacillus to
act)
4- An amount of pre-prepared yoghurt is added to be used as a source of bacteria of yoghurt.
5- Milk is left in an incubator at a suitable temperature, during this period Lactobacillus feeds on the
milk sugar which is known as lactose by respiring anaerobically producing lactic acid and energy.
6- Lactic acid decreases the pH of milk, as a result of this, protein of milk coagulates forming clumps
called curds. In addition, lactic acid gives a sour taste.
7- Yoghurt is then preserved in a refrigerator, to stop the action of bacteria, otherwise it becomes very
sour due to the production of more lactic acid.
Advantages of yoghurt:
1- Has a high nutritive value.
2- Easily digested and absorbed.
3- Tasty, without any additives.
4- Easy to be prepared.

2- Using microorganisms in cheese production:

same steps as yogurt making but rennin( an enzyme extracted from the stomach of calves but nowadays
obtained by genetic engineering)

3- Using microorganisms in bread making:

1- Flour and water are mixed together to form a dough, then yeast is added.
2- The dough is left in a warm place to keep the temperature suitable for the activity of yeast.
3- Flour contains starch, enzyme called amylase and a protein called gluten.
- In presence of water amylase in flour becomes active and acts on starch to be digested into simple
sugar.
- Sugar is absorbed and used by yeast to respire producing carbon dioxide and alcohol (ethanol).
- Gluten forms sticky stretchy threads and this helps the dough to trap carbon dioxide making the
dough rises well.
4- The dough is then shaped into loaves and left again in a warm place to rise.
5- The dough is then baked and this causes:
- Yeast to be killed stopping its action.
- Trapped carbon dioxide in the dough to expand making the bread porous with light texture.
- Ethanol to evaporate.

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4- Using microorganisms in beer and wine making:

- It is the process of conversion of sugar into alcohol and carbon dioxide by yeast (by
means of anaerobic respiration)
1. In Alcoholic drinks
A. Beer:
1. Barley grains are soaked in water for 2 days to germinate and to
activate enzymes in the seeds to digest starch into maltose.
2. The germinated seeds are dried, crushed and mixed with warm water.
3. Yeast is dissolved in the warm water and mixed with the grains.
4. The liquid is covered to allow anaerobic respiration to breakdown
maltose into alcohol and CO 2.
5. CO 2 gives the beer its fizzy sensation.
B. Wine:
- Similar way like beer is used but the sugar comes from grapes instead of
barley.

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SINGLE CELL PROTENS (S.C.P)

(MYCOPROTEIN)
Concept of single cell protein:
Microorganisms such as bacteria, algae, and single cell fungi can reproduce rapidly (asexual
reproduction), their cells contain high proportion of proteins, therefore many types can be used as food.
Method of production of single cell protein
1- Large steel vessels known as fermenters are sterilized to avoid the presence of any microorganisms
that may harm the process.
2- Samples of bacteria, fungi, algae, phytoplanktons or any non harmful microorganisms are introduced
into the fermenter.
3- Nutritive materials are added such as:
a- ammonia as a source of nitrogen and hydrogen for making protein.
b- methanol as a source of carbon and energy.
c- water because most of the cell components are made of water.
d- salts such as sulphur which is necessary for making proteins and nucleic materials.
e- air: contains oxygen for aerobic respiration to produce enough energy for rapid reproduction, also
when air is forced, it pushes the microorganisms keeping spaces between them to increase the surface
area for obtaining the different nutrients.
(Nutritive materials have to be sterilized to avoid the entrance of any microorganisms that may harm the
process.)
4- The products are taken to be used.

Notice:
1- To use green microorganisms such as algae which carry out photosynthesis:
a- light is needed therefore the fermenter has to be transporter or artificial light has to be used.
b- methanol becomes of no need because algae can obtain energy from light and carbon in the form
of carbon dioxide from air during the process of photosynthesis.
c- The process can continue for a long time without adding any new microorganisms because
microorganisms reproduce rapidly inside the fermenter.
Advantages of single cell protein
1- Rapid process leads to the production of large
Amounts of proteins.
2- low fat content so decreases risk of heart
diseases
3- Cheap because waste materials such as left over
Of ammonia are used as nutritive materials for the
microorganisms.
4- can be eaten by vegetarians
5- The produced protein can be stored as powder.
6- The process does not need large place or a soil.
7- independent on weather conditions
8- high protein and fiber content
9- production is always of same quality
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Difficulties facing single cell protein production

1- Eating food which is bacteria or fungi is not
accepted by most people.
2- S.C.P has a little taste and no smell therefore
additives have to be added
3- Many types of SCP have bitter taste due to the presence
of high proportion of DNA and RNA.
4- Many governments and organizations insist that
extra tests should be done on the S.C.P before
being used, and this increases the coast, therefore
most SCP is sold as fodder for animal.

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Fermenter
What is a fermenter?
A vessel in which microorganisms such as fungi or bacteria are grown under controlled conditions in
order to produce substances such as alcohol, antibodies or food substances.
Examples:
- A glass jar used to make wine using yeast is a fermenter.
- A large steel vessel used to grow genetically modified bacteria to produce insulin.

Industrial fermenter
Large tanks usually made of steel, they are equipped so that different environmental conditions can be
controlled such as temperature, oxygen and carbon dioxide concentration, pH and nutrients.
How it is used?
- It is sterilized before starting a process.
- This takes place by introducing very hot steam under high pressure.
Why sterilization using steam instead of using other chemicals or detergents?
If chemicals or detergents are used, their remains may harm the useful organisms which are required to
be grown in the fermenters but steam when cooled it forms water which will not harm the
microorganisms when added to the fermenter.
Importance of sterilization
To avoid the presence of any foreign microorganisms that can compete with the organisms in the culture
reducing the yield of the product, and cause the product to be contaminated with waste products or cells
of the foreign organisms.
Importance of the different parts
1- Inlet:
- For introducing nutrients and other raw materials.
- It is controlled by a value
2- Outlet
A tube for obtaining the products.
3- paddles
- For agitation or stirring, to mix up the contents so that microorganisms can be more exposed to the
nutrients.
- To keep the temperature even throughout the vessel.
- Some fermenter use jets of air to mix the contents instead of paddles.
4- Air inlet
- Through which air can enter for aerobic respiration of microorganisms.
- Air pass through sterilizer or filters to avoid contamination by foreign particles or microorganisms.
5- Temperature and pH probes
- Temperature probe to detect the temperature, so that it can be adjusted.
- pH probe to detect the changes in pH so that it can be adjusted by adding acid or alkali to provide the
optimum pH.

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How to adjust temperature in the fermenter

- Temperature may increase as a result of metabolic activities of microorganisms in the fermenter, this
increase can be detected by temperature probe.
- To reduce temperature to its suitable value, cold water is forced into the water jacket until reaching
the required temperature.
Notice:
Stainless steel or special alloys are used in making fermenters because many microorganisms produce
acidic substances that have corrosive action on the walls of the fermenter.

Using fermenter (bioreactor) to manufacture enzymes

The organisms used
Yeast, fungi or bacteria.

Method
1- Feedstock such as starch is sterilized and is forced into the fermenter to be used by microorganisms
which are allowed to grow producing their enzymes.
2- If the enzymes are extracellular, they are extracted directly from the filtrate.
3- If the enzymes are intracellular, the microorganisms are filtered out of the feedstock to be crushed and
then the enzymes are extracted with water or other solvents.
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Role of Penicillium in the production of Penicillin

-Antibiotics are substances that kill bacteria without harming human cells. Antibiotics are used to cure
bacterial infections.
- Penicillin is an antibiotic made by growing the fungus Penicillium in a large fermenter.
- The fermenter contains carbohydrates and amino acids.
- The contents are stirred continuously to
1- Keep fungus in contact with nutrients
2- Mix oxygen into culture
3- Roll fungus up into pellets which makes it easier to separate the liquid part of the culture (which
contains penicillin) from the fungus at a later stage.
- The rate at which fungus produces penicillin depends on how much sugar is available. If there is a
lot of sugar or no sugar at all penicillin will not be produced. Only small amounts of sugar have to be
fed into the fermenter.
- Bubbles in the fermenter provide oxygen and mix the nutrients and penicillium together.

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