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Enzymes

 Derived from Greek World ( En = in, Zyme = Yeast )


 Enzymes are commonly proteinaceous substances of high molecular
weight which accelerate, or catalyze, chemical reaction of biological
origin without them self undergoing any change.
 Enzymes have a high degree of specificity for types of reaction
catalyzed and for their substrate
 The term enzymes was coined by Kuhne in 1878
 First enzyme extract from yeast cell by Buchner (1897)

Enzyme nomenclature and classification


Nomenclature
 Previously enzymes use to be named by adding ‘in’ e.g. Pepsin,
trypsin etc.
 In 1883 Duclaux proposed the naming of enzymes by adding the
suffix "-ase" to the substrate on which the enzyme acts or the
chemical reaction it catalyzes. Example enzymes acting on sucrose as
sucrose, enzymes catalyzing oxidation oxidase.
 The International Union of Biochemistry and Molecular Biology have
developed the EC ( Enzyme commission) numbers of four digits
Preceded by “EC”.
First digit denotes: Class
Second digit denotes: Sub class
Third digit denotes: Sub-sub class
Fourth digit denotes: Sub-sub-sub class
For examples example, hexokinase (EC 2.7.1.1) is a transferase (EC
2) that adds a phosphate group (EC 2.7), a molecule containing an
alcohol group (EC 2.7.1) to a hexose sugar (EC 2.7.1.1)

First two digits represents type of reaction it catalyzed, last two digits
represents substrate which enzyme acts.

Properties of enzymes
1. Enzyme is specialized protein
It exhibit all properties of proteins
2. Colloidal nature : Due to large size of molecules
3. Catalytic property
Enzymes are very efficient catalyst. Only small amount of
enzymes is enough to convert large quantity of substrate in
product. It Speed up reaction by 106 - 1012 times greater than
those of the corresponding un catalyzed reactions
4. Specific
Enzymes are more specific toward their substrates and for the type of
reactions that catalyze.
Enzymes shows 3 types of specificity
a. Specific specificity
Some enzymes act on only one substrate
e.g. Urease acts only on urea.

b. Group specificity
Catalyze the reaction of structurally related groups only
Amylase hydrolyses the group of substances like starch, dextrin and
glycogen, which have the same type of glycosidic linkages (α1,4).

c. Optical Specificity
The enzymes acts on only one of the two optical isomers of a compound
For example L-amino acid oxidase acts only on L-amino acid but not on
its D-form of amino acid.

5. Heat sensitivity ( note )


Enzyme is very sensitive to heat. Optimum temperature for enzyme
action is 35-37 degree centigrade’s. The enzymes activity decreases
below or above this temperature due to denaturation.

6. pH sensitivity
Enzyme is dependent upon the pH of where the reaction is taking
place, e.g. pepsin in the stomach has an optimum pH of about pH2,
Whereas salivary amylase has an optimum pH of about 7. 4.
7. Reversibility of reaction
Enzymes are capable of bringing reversion in a chemical reaction

Function of enzymes
1) To accelerate or retard or bring about reaction
2) Regulate reaction
3) To make possible the metabolic reactions
4) To facilitate reaction
5) To break down larger molecule to small molecule
6) To carry out flow of reaction smoothly

Mechanism of enzyme action


Enzymes helps in the progress of the reaction in 2 ways
1. Lowering activation energy

2. Formation of enzyme substrate complex


There are three theories to explain formation of enzymes substrate
complex.

a. Michaelis-Menton Hypothesis

The theory postulates that the enzymes has number of active sites where
the substrate is bounded to form weak enzyme substrate complex (S).
This enyzme-substrate complex, on hydrolysis, decomposes to yield the
reaction product (P) and the free enzyme (E). These reactions may be
symbolically represented as follows:
[S] + [E] ----------> [ES] ---------> P+[E]

b. Fisher’s lock and key model


This model was proposed by Emil Fischer in 1898. According to this
model, the union between the substrate and the enzyme takes place at the
active site more or less in a manner in which a key fits a lock and results
in the formation of an enzyme substrate complex. The enzyme-substrate
complex is highly unstable and almost immediately this complex
decomposes to produce the end products of the reaction and to
regenerate the free enzyme.

c. Koshland’s Induced fit model


• Enzyme structure flexible, not rigid
• Enzyme and active site adjust shape to bind substrate
• Increases range of substrate specificity
• Shape changes also improve catalysis during reaction

Natural polymers such as cellulose, starch, rubber, wool, silk, etc.


BIODEGRADABLE POLYMERS
• Biodegradable polymers are defined as polymers comprised of
monomers linked to one another through functional groups and have
unstable links in the backbone.
• They are broken down into biologically acceptable molecules that are
metabolized and removed from the body via normal metabolic
pathways.
• Based on biodegradability polymers are classified as:
1. Biodegradable polymers eg: collagen, poly glycolic acid etc.,
2. Non biodegradable polymers eg: poly vinyl chloride, polyethylene
etc.,

POLYLACTIC ACID

• Polylactic acid or polylactide (PLA) is a thermoplastic aliphatic


polyester derived from renewable resources, such as corn starch, tapioca
products (roots, chips or starch) or sugarcane.
• It can biodegrade under certain conditions, such as the presence of
oxygen, and is difficult to recycle.
• Highly crystalline, high melting point, low solubility.

• Bacterial fermentation is used to produce lactic acid from corn starch


or cane sugar.
Synthesis:
Lactic acid/Glycolic acid is first polycondensed to yield low molar mass
oligomers. Thesynthesis of homo and copolymer of lactic and glycolic acid is
carried out by the ringopening melt condensation of these cyclic dimers, lactide
and glycolide.
Mechanism:
The lactide/glycolide polymer chains are cleaved by hydrolysis to the monomeric
acid. Theyare eliminated in vivo through the kreb’s cycle, basically as carbon
dioxide and urine. Thedegradation rate at various body sites is more or less
identical as the hydrolysis of thesepolymers is dependant only on significant
changes in temperature and pH and presence of catalyst.The biodegradation of
lactide/glycolide polymers primarily occurs in two steps. First ,random hydrolytic
cleavage of the ester linkage leading to the reduction in molecular weightand
second, the onset of weight loss and a change in the rate of chain scission.
Application:

The lactide/glycolide polymers are basically low melting thermoplastics. These


offersconsiderable flexibility and ease in fabrication of lactide/glycolide polymer
baseddrug delivery system, like microparticles, implants and fibres.

Lactide/glycolide based microspheres can also be prepared. (by solvent


evaporation,phase separation and fluidized bed coating)

The lactide/glycolide polymers can be fabricated into mono- or multifilaments.

APPLICATIONS • PLA is used in the preparation of sutures or


orthopaedic devices.

FACTORS AFFECTING BIODEGRADATION OF POLYMERS


 Morphological factors
• Shape & size
• Variation of diffusion coefficient and mechanical stresses

 Chemical factors
• Chemical structure & composition
• Presence of ionic group and configuration structure
• Molecular weight and presence of low molecular weight compounds

 Physical factors
• Processing condition
• Sterilization process

Waste Management

Solid waste can be classified into different types depending on their


source:
a) Household waste is generally classified as municipal waste,
b)Industrial waste as hazardous waste
c) Biomedical waste or hospital waste as infectious waste.

Thermoplastic Olefin, or TPO is a combination of polymer and filler


blends typically consisting of thermoplastic material, such as block
copolymer polypropylene, an elastomer or rubber, and mineral filler
such as calcium carbonate or talc. This material is ideal for applications
for industrial markets where material durability for long-term use are
integral to part function.

This material PO is frequently used in interior and exterior automotive


applications as a replacement for more traditional materials such as
metal and other engineering thermoplastics. As a result, TPO needs to
be able to withstand harsh weather conditions and exposure to UV light.
Today there are many grades of TPO available that offer UV protection
through the use of special additive packages that prevent custom colors
from fading due to sun exposure over an extended period of time. In
addition, this material provides the optimum balance of stiffness, cold
temperature impact, and low thermal expansion, making it ideal for
applications where weatherability and durability to all conditions is
crucial.

These qualities are perhaps the leading reason why this polymer has
found a home in Automotive and Industrial markets. In both of these
markets, the finished part will have exposure to heat and cold, and will
need to keep its shape while allowing for limited thermal expansion or
contraction. Imagine if on a hot summer day the bumper on car started to
expand and cracked! As automotive companies began to shift from
traditional metal materials to materials like TPO, avoiding problems like
this was at the top of their list of requirements when qualifying
replacement materials.

TPO sheet can be laminated with decorative and paint films achieves the
durability and aesthetics required to maintain the high quality surface in
applications where scratch and mar resistance is critical. Our extruded
TPO sheet is ideal for thermoformed parts used in demanding
applications such as:

 Household appliances
 Interior and exterior automotive parts
 Industrial equipment
 Recreational vehicles

Biodegradability is very important as a parameter of the environmental


behaviour of a substance. Among the methods used to determine the
biodegradability of an insoluble substance, the Soil Burial Test (SBT)
has been proposed as also suitable for use with plastic polymers. this
paper reports on year long SBT's which were carried out in garden soil
on four plastic polymer samples: three with starch and/or fatty acids
added as biodegradation promoters and one unadulterated reference
sample. The samples were buried in garden soil. The addition of
promoters did not result in degradation in all cases.
7.5. Soil burial test
Tests based upon this methodology evaluate in-service soil contact
exposure. The material is buried in soil beds prepared in the laboratory
using standard sieved soil; often a commercial soil-based compost. The
soil beds are normally conditioned for up to four weeks prior to use
and may be supplemented with organic fertilizer to encourage an active
microbial flora. The microbial activity is tested using a cotton textile
strip which should lose 90% of its tensile strength within 10 days of
exposure to the soil. Currently no other reference materials are
recommended, although for plastic materials a standard alternative able
to demonstrate the degradation capabilities of the microbial flora with
respect to plastic should be sought. The soil beds containing the samples
are incubated at a constant temperature for between 28 days and 12
months. The moisture content is normally set at 20–30%, although it is
better calculated as a percentage (40–50%) of the soil’s maximum water
holding capacity. This then accounts for different soil structures and
ensures that the soil does not become unduly wetted or is too dry for
optimal microbial activity. Samples are removed for assessment of
changes or a light microscopy and SEM examination to assess surface
damage and to look for the presence and nature of microbial growth.
Physical factors such as fragmentation and embrittlement can also be
assessed in these tests. Finally, the samples can be used to ‘bait’
microorganisms involved in the degradation process. These microbes
once isolated and characterized can be incorporated into the petri dish
screen as alternatives or additions to
the current list.