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Pushneet Sahdra

Tejaswita Sachdeva

• History

• Introduction

• Understanding enzymes

• How it works?

• Work use areas

• Production of enzymes

• Composition of an enzyme detergent

• Protein versus genetic engineered enzymes

• Detergent enzymes





• Miscellaneous detergent enzymes

• Enzymes formulation

• Production of enzyme based detergents

• Enzyme stabilization

• Application of enzyme based detergents

• Benefits of enzymes

• Conclusions

• Bibliography
What is an enzyme?

picture 1

picture 2

Enzymes are proteins, composed of hundred of amino-acids, which are produced by

living organisms. They are responsible for a number of reactions and biological activities
in plants, animals, human beings and micro-organisms. They are found in the human
digestive system to break down carbohydrates (sugars), fats or proteins present in food.
The smaller pieces can be absorbed into the blood stream.

Each enzyme is a made of a sequence of amino acids (like pearl on a string, picture 1)
folded into a unique three-dimensional structure that determines the function of the
enzyme. Even the slightest change in the sequence of the amino acids can alter the
shape and function of the enzyme.

Only a small part of the enzyme participates in the catalysis of biochemical reactions:
the active site (picture 2). Enzymes are therefore very specific (e.g. a cellulose can only
degrade cellulose).

Enzymes are essential for all metabolic processes, but are not themselves living
materials. They are distinguishable from other proteins because they are known as
biological catalysts (substances which speed up reactions but which do not get used up

Enzymes are proteins created by living cells that exist in organisms such as plants, animals and bacteria
and are used to digest waste. When added to organic material like dirt, grease and oil, they immediately
go to work breaking down the organic material within these substances. This natural “dust to dust”
process that constantly occurs in our environment keeps waste material from overrunning us. The four
basic enzymatic systems are those that break down fats and greases (lipase); proteins (protease);
cellulose such as wood, cotton and paper (cellulase); and carbohydrates and starches (amylase). Dirt has
layers of fine film composed of “substrate” such as grease, oils, fats, bacteria, germs, dust mites, non-
organic material and organic microorganisms. These films are bonded to each other and to the surface by
amino and fatty acids (organic acids composed of proteins, fats or fatty oils). Most cleaners emulsify
some of these dirt films but may not break down the lower levels held together by amino and fatty acids.
Usually the top layers of the films are removed but some of the lower levels are left to collect bacteria. As
a result, re-soiling can occur much faster.


When activated, enzymes attack or digest the amino and fatty acids that bond the films of dirt together.
They also emulsify them so they can be completely removed from the surface. Researchers believe that
in the activation process, when the substrate and enzymes come in contact with each other, the enzymes
physically curl and twist—in what is called a “conformational change.” This physical change initiates the
contact between the enzyme and substrate which is necessary to “catalyze” the reaction. A catalyst is a
substance that speeds or slows a chemical reaction without being involved in the reaction itself. Put
another way, enzymes are chemical catalysts that accelerate the natural biodegrading, or breaking down,
of organic substrate, which comprises most soils. Enzymes dissolve and break down protein and organic
matter, diminishing odors caused by staining agents such as urine, feces, vomit, pet odors, spoiled foods
and mildew. Enzymes are derived from living organisms and are harmless to humans, animals, marine
life and general ecology. They are non-toxic, non-irritating, non-gaseous, non-flammable, non-pathogenic
and typically safe to use. There are thousands of different enzymes, each having specific, individual
characteristics. Since an enzyme that breaks down proteins (protease) will not react on fats or oils, and
effective enzymatic cleaning system must contain enough different classes and types of enzymes to
assure proper catalytic reaction. In concentrated form, this greatly speed up the natural “dust to dust”
process. One way to demonstrate the effectiveness of enzyme digesters is to mix warm water and the
enzyme product in a small cup (per recommended dilution ratio). Then place a few pieces of dry cat food
into the cup. After 10 to 15 minutes, the cat food will be totally dissolved. This breakdown of protein will
demonstrate, and help you more fully understand, how the chemical works on other microscopic bacteria
and proteins.


Drain Openers. Follow label for correct mixing instructions, then pour into clogged drains Always start by
working on lower level floors drains first. If your workers start at the upper levels, the dislodged and
dissolved protein will further plug lower level plumbing. It is best to use drain openers at night or over a
weekend to give the enzyme several hours to do its job. Enzymes are not fast acting like acid-type drain
openers and they require a few hours to work properly; however, they are much safer for workers and

Carpets. Enzymes work well for blood stain removal and they are very effective in reducing (or in most
cases, eliminating) odors caused by urine, vomit and other organic-related odors. When odors are in
carpet backing, use a carpet syringe and inject 1 ounce of undiluted enzymes through the backing onto
the sub-floor. Several injections are required to cover a large area. Each injection should cover a 3-foot
diameter area. Enzymes can be used on all other water-safe fabrics that contain odor or stains caused by
the same organic matter that also stain carpeting.

Restrooms. When mopping, mix enzymes with warm water to the correct dilution ratio and mop floors.
Do not rinse floors, but air dry, allowing the enzymes time to react with bacterial matter. Enzymes will be
absorbed into the floor mortar joints, allowing deep odor removal. Remember that you cannot use an
enzyme digester at the same time that a disinfectant cleaner is applied. The residue of the disinfectant will
kill the live organisms of the enzymes. Use one or the other—never both digester and disinfectant
together. You can also spray enzymes on and around urinals and other odor-producing fixtures. Regular
applications of enzymes will eliminate the source of the odor. When spraying, use a stream, not a mist.
Enzymes applied as a mist can easily be inhaled into the lungs. Enzymes are living organisms and could
cause medical problems if inhaled.

Cost Effectiveness. Enzymes are not costly; however, care should be taken to correctly use these
products. You need to identify specific areas that are present or potential problem areas. You can then
work the enzymes into your present program at proper frequencies to ensure desired results. Using
enzymes in a haphazard manner with no scheduled routine will not only waste product, but also more
importantly will waste valuable labor. Enzymes are economical and safe to use within a wide work-use
area. They can be injected directly into mattresses to reduce urine smells or poured into kitchen or
restroom drains to unclog grease or hair deposits. The primary caution is not to permit inhalation of
sprayed (misted) product. Some good candidates for bacteria / enzymes digesters are health care
facilities, schools, industrial plants, health clubs, correctional facilities, offices, restaurants, and food
service operations. The list can be expanded to include any and all areas that have stains and odors from
protein or organic matter. Work use areas for enzymes are expanding, and new applications open up
daily in the sanitary maintenance field. Enzymatic cleaners are new technology that allows for many uses
by cleaning professionals

Production of enzymes
Enzyme molecules are far too complex to synthesize by purely chemical means, and so the only
way to make them is to use living organisms. The problem is that enzymes produced by micro-
organisms in the wild are often expressed in tiny amounts and mixed up with many other
enzymes and proteins. These micro-organisms can also be very difficult to cultivate under
industrial conditions, and they may create undesirable by-products.

Modern industrial cultivation of enzymes begins with fermentation of a vial of dried or frozen
micro-organisms called a production strain. This production strain is selected to produce large
amounts of the enzymes of interest. The production strain is first cultivated in a small flask
containing nutrients and agar. The flask is placed in an incubator which provides the optimal
temperature for the previously frozen or dried cells to germinate. Once the flask is ready, the
cells are transferred to a seed fermenter, which is a large tank containing previously sterilized
raw materials and water, known as the medium. Seed fermentation allows the cells to reproduce
and adapt to the environment and nutrients that they will encounter later on. The cells are then
transferred to a larger tank, the main fermenter, where temperature, pH and dissolved oxygen are
carefully controlled to optimize enzyme production. Additional nutrients may be added to
enhance productivity. When main fermentation is complete, the mixture of cells, nutrients and
enzymes, referred to as the broth, is ready for filtration and purification.
Protein versus genetic engineered enzymes

Protein engineering is technique used to alter the gene encoding for an enzyme in order to
change or obtain new properties. The genetic integrity of the organism producing the enzyme is
not changed.

Each enzyme consists of several hundreds of amino acids located in such a delicate three-
dimensional structure. This structure determines the properties of the enzyme such as reactivity,
stability and specificity. Based on protein engineering, scientists can construct slightly altered
enzymes by modifying the gene encoding for the enzyme.

Engineered organisms then produce the modified enzyme which is subsequently tested to
evaluate whether the structure/function models have been correct. Such innovative methods have
led to the discovery of detergent enzymes which are much more active, efficient and / or robust
in terms of pH, temperature and / or chemical stability ( e.g. vs bleach).

Genetic engineering is the alteration of the genes of the organisms. The genetic integrity of the
organism producing the enzyme is changed for ever. Usually such enzymes are used in a
confined environment and are sometimes engineered in a such way that they can not survive in
the natural environment.Such alterations can be effected by breeding and by mutation - the
natural processes that for billions of years have formed the basis for the evolution of new
organisms. The process whereby genes mutate to achieve small (but sometimes beneficial)
alterations is called mutagenesis.

P&G now can screen for wild-type microorganisms / genes in line with the identified consumer
need, e.g. an alkaline cold wash enzyme. "Compressing" the above mentioned natural evolution
process into a short-term period, i.e. a limited number of cycles of directed evolution is a major
challenge in using mutagenesis to improve a strain of micro-organisms / a gene. Key is to start
from the right substrate screen. Thousands up to millions of mutants therefore have to be tested
to find the optimal strain. Nowadays, this classical method of screening micro-organisms for
beneficial mutations has become a high-tech process. To be able to test this massive amount of
mutants, which are only available in microgram quantities, efficient automatic system are
available which are capable of simultaneously scanning dozens of plates filled with mutant
strains of micro-organisms. Without the need for human intervention, robots measure the enzyme
activity produced by the individual mutants in a highly efficient manner. What once took years is
now achieved in a few days. Such robots are capable of discovering hundreds of new, interesting
mutant strains of micro-organisms / genes that research scientists can further test and
characterize using other systems.

12. www.Chemical Function Definitions -

13.www. Enzyme and Bacterial Cleaning

14.www. Enzymes in washing powders Biotech Learning Hub

15. www.Understanding Enzymes Used In

16.Book- Chemistry Of the Textile Industry(page-157-160)