Sukhkaran Dhaliwal AP Biology Thursday, October 9, 2008

Applications of Enzyme Use

1.
Pickling, which is also known as brining or corning is the process of preserving food by
anaerobic fermentation in brine, a solution of salt in water, to produce lactic acid, or marinating
and storing it in an acid solution; the most common solution being vinegar (acetic acid). Pickling
is unique and beneficial because the solution has a pH less than 4.6, which is adequate to kill
most bacteria. The food is placed in an edible anti-microbial liquid that inhibits or kills bacteria
and other micro-organisms that could spoil food. If the food contains sufficient moisture, a
pickling brine may be produced simply by adding dry salt. Natural fermentation at room
temperature by lactic acid bacteria produces the required acidity. Unlike canning, pickling
doesn’t require that the food be completely sterile before it is sealed.
Bacteria, yeasts, and moulds need a food source and water to grow. When food is
dehydrated, most of the water is removed which prevents this growth process. Then it can be
said that by reducing water activity sufficiently, it can be delay or prevent bacterial growth.
Dehydrating also slows down the action of enzymes in food. Enzymes are naturally present in
food, causing it to ripen and eventually spoil. Therefore, by slowing down enzyme action, food
will last longer. When water is removed, enzyme action is not completely stopped. The removal
causes the enzyme to become inactive until it is hydrated again.
Canning foods essentially means boiling them then putting it in can (while still boiling or
after it has finished). A can be any sealable container. When you boil the food, you are killing
the bacteria and causing the enzymes to become inactive. The heating and later cooling forms a
vacuum seal, which prevents other microorganisms from contaminating the food once within
the can again. The can in which the food is in is sterile and has no bacteria because of it boiled.
Once the can is opened, the vacuum seal is broken, allowing bacteria to attack the food. This is
why some products give instructions to refrigerate the contents after opening.
The principal of freezing food for preservation is based largely on the fact that reducing
the temperate of the food lessens the activity of the enzymes to a minimum. The purpose of
freezing the food is so the enzymes will not be able to live, which is true in most cases, but some
of them may still live, but have almost no activity. The effect is negligible and will therefore
cause food to last longer. This method of preservation is still one the most used by people.

2. Making yogurt has many steps involved, but before the milk is inoculated with the bacteria, the
milk is cooled to about 40oC after pasteurization and homogenization. During pasteurization,
the milk is heated to about 85oC, and then cooled for inoculation. To be named a yogurt, it must
be made with the bacterial species Streptococcus salivarius subsp. thermophilus and
Lactobacillus delbrueckii subsp. Bulgaricus. The optimum temperature for these bacterial
cultures is approximately 40oC. When the bacteria are added to the milk, it is already at its

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Sukhkaran Dhaliwal AP Biology Thursday, October 9, 2008

optimum temperature so yogurt production can occur much faster. During this time, the
bacterium metabolizes certain compounds in the milk producing the characteristic yogurt flavor.
An important byproduct of this process is lactic acid. The longer the inoculation, the mort tart
the yogurt will be due to more lactic acid being created.

3. Rennin is also known as an enzyme called chymosin. In the case of chymosin, the reaction
involves the breakdown of casein in milk resulting in clot formation in the presence of calcium
ions. This is the basis for the production of many varieties of cheeses. The chief milk protein,
casein, is coagulated by the enzyme action of rennet. Coagulation when used in the context of
cheese production means separating the whey (liquid) from the curds (solid). The specific
linkage between the hydrophobic and hydrophilic group of casein inside would milk would
between broken if chymosin causes cleavage of the peptide bond between phenylalanine and
methionine in the kappa-casein. The hydrophobic group would unite together and would form a
3D network to trap the aqueous phase of the milk. Due to this, rennin is added to bring about
the extensive precipitation and curd formation in the cheese. The temperature of the milk is
kept at 35oC because this is the temperature where the ideal amount of whey is separated from
the curd. If the temperature is higher, it forces more whey out from the curd. This makes sense
because rennin is extracted from the stomach of the calf and the body temperature of a calf is
about 37oC. The optimum pH of the enzyme is approximately 3.5.

4. Starch can be converted to glucose syrups by hydrolysis, and the syrups find many applications
as sweeteners in foods, such as candy, biscuits, jams and jellies, and pharmaceuticals like cough
syrups, tonics, and vitamin preparations. Hydrolysis can be accomplished without enzymes, but
this method can generate distasteful flavours and unsightly colours. Enzymatic hydrolysis offers
a cleaner and more efficient hydrolysis. In a common process, α-amylase, produced by the
bacterium Bacillus licheniformis, hydrolyzes starch to maltose. Then the enzyme glucoamylase,
produced by moulds such as Aspergillus and Rhizopus, is added to hydrolyze maltose into
individual glucose molecules
amylase glucoamylase (maltase)
amylose and amylopectin maltose α- glucose
B. licheiforms Aspergillis or Rhizopus species

5. Lactose intolerance is the inability to digest and absorb significant amounts of lactose, which is a
major sugar that is found in milk. Lactose is made up of two simple sugars called glucose and
galactose (both with the chemical formula C6H12O6), which are isomers of each other. After it is
breaks down into those two components, it is then absorbed into the bloodstream. Those who
cannot break down lactose into its simpler forms are said to be lactose intolerant. For the
lactose to be absorbed in the bloodstream, it must be broken down into the glucose and
galactose in the small intestine. The enzyme that splits lactose into glucose and galactose is
called lactase and is located on the surface of the cells lining the small intestine. Lactose
intolerance is caused by reduced or absent activity of lactase that prevents the splitting of
lactose. Some of the symptoms of lactose intolerance include nausea, cramps, bloating, gas,

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Sukhkaran Dhaliwal AP Biology Thursday, October 9, 2008

and diarrhea. For those react to very small amounts of lactose or have trouble limiting their
intake of foods that contain it, the lactase enzyme is available without a prescription to help
people digest foods that contain lactose. The tablets can be taken with the first bite of dairy
food. Lactase enzyme is also available as a liquid. Adding a few drops of the enzyme makes
lactose more digestible for people with lactose intolerance. Lactose-reduced milk and other
products are available at most grocery stores. The milk contains all of the nutrients found in
regular milk and remains fresh for about the same length of time or longer it is super-
pasteurized.

6. Dirt commonly found on clothing includes proteins, starch, and lipids. Although it is possible to
remove these stains with soaps and detergents, enzymes allow stains to be removed at lower
temperatures and will less mechanical agitation in a washing machine. Enzymes are also more
effective than nonbiological cleaning agents at removing stains such as blood, grass, milk, and
perspiration. The cleaning industry commonly adds proteases and amylases to detergents to
help remove protein and carbohydrates stains from clothing and other fabrics. The enzyme
protease breaks down protein stains and lipase breaks down fats and oils (lipids). The
breakdown of protein and fat molecules is called hydrolysis, which simply means reaction with
water. The enzyme catalyzes these hydrolysis reactions, therefore help with the removal of
stains. Biological detergents and washing powders use lipases and proteases to break down fat
and protein molecules from food stains, which have remained on clothing. The fatty acids,
glycerol, and amino acids are removed in the water during the washing process. Also, cellulases
are use for color brightening and softening of cotton garments. Enzymes are also used in
textiles for the stone washing of denim (in combination with pumice stones), biopolishing and
softening of cotton, bleaching, cleanup, and removal of starch from woven materials.

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