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Group II TBM Class

2314100095 M.Aziz Rahmatullah
2314100135 Dany Destyawan Anwar
2315100047 Surya Aji Prasetya
02211748007002 Fatin Fatihah
• Proteins are polymers of amino acids linked
together by peptide bonds.
• Two or more amino acids joined together are
called a polypeptide, while molecules that consist
of one or more polypeptides are termed proteins.

The general structure of amino acids.

• The general structure consists of an amino group (NH2), a carboxyl group

(COOH), a hydrogen atom and a distinctive R group all bonded to a single
carbon atom,called the α-carbon.
• A peptide bond is formed between the α-carboxyl group and the α-amino
group of two amino acids by a condensation (or dehydration synthesis)
reaction with the loss of water.
• The ionization state are varies with pH.
Neutral pH : Amino acids are predominantly zwitterions.
Acidic pH : The carboxyl group is un-ionized and the amino
group is ionized
Alkaline pH : The carboxyl group is ionized and the amino group is un-
• Due to R group differences, the 20 amino acids can be divided into four
categories: non-polar, polar, acidic and basic amino acids.
• The charge state of an ionizable molecule is dictated by its pK
value, the pH at which an equal number of protonated and
deprotonated species exist resulting in a net charge of zero.
• If pH is above pK of a particular molecular species then the
molecules will be deprotonated and if pH is lower than pK then
the molecules will be protonated.
• The pH at which a protein has zero net charge is called pI or the
isoelectric point.
• Some proteins contain non-standard amino acids in addition to
the 20 standard amino acids.
• These are formed by modification of a standard amino acid
following its incorporation into the polypeptide chain (post-
translational modification).
• Food proteins have an important nutritional role and are
primarily used by the body to supply nitrogen and amino
acids from which the body synthesizes its own proteins.

Amino acids can be categorized by 2 types which is :

1. Essential Amino Acids
Amino acids that your body cannot create on its own.

2. Non-Essential Amino Acids

Synthesized by the body.
• Contractile Proteins
• Structural proteins
• Storage proteins
• Transport proteins
• Antibodies
• Enzymes
• Hormonal proteins
The protein requirements of humans are
dependent on a number of factors:
• Gender.
• Age.
• Body weight and composition.
• Energy intake.
• Micronutrient composition of the diet.
• Weight Gain / obesity
• High blood pressure
• Heart attack / Stroke
• Reduced liver and brain function
• Stone in a kidney
• Dehydration
• Hypoglycemia
• Low blood pressure
• Anemia
• Wounds take longer time to heal
• Illness of liver and kidneys
• Stomach ulcers
• Nerve weakness
• Bad circulation
• PEM – Kwashiorkor and Marasmus
Protein Synthesis
• Transcription is the synthesis of a
complementary strand of RNA from a DNA
• Messenger RNA (mRNA) carries the coded
information for making specific proteins from
DNA to ribosomes, where proteins are
• Translation is decoding the language of nucleic
acid and converting that information into the
language of proteins
• The codons of mRNA are converted into protein
through the process of translation. The codons of
an mRNA are read sequentially
• The site of translation is the ribosome, and
transfer RNA (tRNA) molecules both recognize
the specific codons and transport the required
amino acids
The Genetic Code

The three nucleotides in an mRNA codon

are designated, respectivety as the first
position, second position, and third position
of the codon on the mRNA.

Each set of three nucieotides specifies a

particular amino acid, represented by a
three-letter abbreviation.
The 20 Amino Acids Found in Proteins
Protein Structure
• Primary Structure refers to the amio acid sequence
• Secondary structures are the structures formed by amino acid
chains, e.g. α-helix, β-sheet, random coil
• The tertiary structure of a protein refers to the way in
which secondary structures, e.g. helices, sheets, and
random coils, pack together in three dimensions
• Quaternary structure refers to the association of tertiary
structures, e.g. two associated subunits, each a separate
polypeptide chain
Protein Denaturation
• Denaturation is a phenomenon that involves transformation of a
well-defined, folded structure of a protein, formed under
physiological conditions, to an unfolded state under non-
physiological conditions.

These are the consequences of Denaturation:

• Loss of biological activity (e.g. enzyme activity).
• Loss of solubility
• Increased intrinsic viscosity.
• Increased susceptibility to proteolysis.
• Improved of digestibility
• Changes in texture
Reversible Denaturation and Irreversible Denaturation
Behavior of Denatured Protein

Hydrophobic core

Hydrophilic surface
Fast under non-physiological conditions

Slow under physiological conditions


Unfolding forces some

hydrophobic AA to
or other ingredient interactions
There are various denaturing agents that can destabilize protein structures
that are categorized as physical agents and chemical agents.

• Physical agents include heat, mechanical treatment, hydrostatic

pressure, irradiation, and adsorption at interfaces.

Heat denaturation is useful in food processing since it tends to lead to

improvement of sensory properties and protein digestibility, and can be used
to manipulate foaming and emulsifying properties.
• Chemical agents to denature proteins include acids, alkalis,
metals, organic solvents and various organic solvents.

Exposure to acids or alkalis (i.e. pH changes) affects the overall net charge on a
protein, which will change the extent of electrostatic interactions, both
attractive and repulsive. Most proteins are stable within a pH range around
their isoelectric point (zero net charge) and the effects of acids or alkalis are
normally reversible.
• Heavy Metal salts
Usually contain Hg, Pb, Ag, TI, Cd, and other high atomic weights. Since salts are ionic they distrupt
salt bridges in protein. The Reaction of a Heavy metal salt with protein usually leads to an insoluble
metal protein salts. As for example AgNO3 is used to prevent gonorrhea infection.

• Detergents
Hydrophobic parts of detergent associate with hydrophobic parts of the protein which dissociating
hydrophobic parts of protein that no longer associating with other that can lead to unfolding of
protein chain.

• Organic Solvent
weakens hydrophobic interactions since non-polar side chains become more soluble. Organic
solutes can have a variety of effects. Urea alters the structure of water in such a way as to weaken
hydrophobic interactions, leading to protein unfolding
Protein Purification
The critical study of food protein structure and function
is the ability to purify protein. Separating a particular
protein from a complex mixture of many proteins, often
hundreds or thousands, is done by taking advantage of
differing biochemical characteristics such as charge, pI,
mass, molecular shape and size, hydrophobicity, ligand
affinity, and enzymatic activity among others.

Two critical categories of purification techniques

typically employed in protein purification schemes are :
1. Ion Exchange
2. Size Exclusion.
In Practical Term of purification, Chromatography is the
most important technique used.

Preparative Chromatography involves the separation of

protein (or other substances) dissolved in a mobile phase
based on their differecential interractions with a stationary

The Mobile phase = can be hydrophobic ( organic solvent) or

hydrophilic (aqueous buffer).
The stationary phase = can be hydrophobic or hydrophilic
and / or an immobilized ligand or molecule with specific
affinity for proteins.
The simplest example of chromatography ion exchange
where the stationary phase consists of charged molecules
immobilized on a solid polymer packed into a column.
1. If the target protein is anionic at a given pH then
passing a protein mixture through a stationary phase
containing positive charges will result binding and
retention of the target protein. While cationic and
neutral proteins will flow through the anion exchange
2. The bound anionic proteins can be eluted from the
column by introducing increasing concentrations of
anions (eluent) that compete for the positve charges of
the stationary phase.
3. Weakly bound proteins will elute at
lower concentrations of eluent and
strongly bound proteins will elute at
higher concentrations of competing
eluent, thereby separating protein
based on their respective binding
affinities for the stationary phase.

Very large quantities to very small

amounts of protein can be separated in
single ion exchange runs depending on
the equipment scale chosen.
In the case of size exclusion chromatography, column
packing consists of inert, spherical packing material
(beads) containing specific pore sizes.
1. The Highest molecular mass of protein capable
pasing through the pores is termed the
exclusion limit.
2. Protein of mass in excess of this limit may only
pass around the packing material, thus taking
the shortest path thorugh the column

Picture from : size excluison chromatography principle and

methods handbook

3. While the smaller proteins that enter and exit beads take a more convoluted
longer path.
4. Hence larger proteins elute before smaller proteins in size exclusion
chromatography, thereby allowing for the separation of many proteins within a
mixture based on size.
The resolution for separation of masses varies with :
1. Quality of setup purchased
2. Length of the column.

Generally, only milligram quantities are separated by this

chromatoghraphies technique.

Filtration device with spescific pore size e.g. ultrafiltration can be

used for bulk separation of protein where only two fractions are
obtained, flow-through and retentate, yet with the advantage of
processing relatively large quantities of protein per run.
Protein Analysis Techniques
The most basic type of protein analysis is that of
determining the total content of substances.

This measurement is based on the average nitrogen

content of protein, 16 %. Organic nitrogen reacts with
sulphuric acid when heated to give ammonium

This product is converted to ammoinum hydroxide

which can be quantified by titration, indicating the
quantity of nitrogen present and hence the approximate
amount of protein in a food.
To determine the size of protein can be use electrophoresis.
Or It also can use technique Poly Acrylamide gel electrophoresis (PAGE).

1. Briefly, when proteins are passed through

a polymerized acralamide gel, smaller
proteins will travel more quickly than
larger proteins, allowing for a means for
their separation.
2. The motive force that makes proteins pass
through the gel is provided by applying a
voltage across the gel such that negative
molecules migrate down the gel toward
the anode.
3. If charge amounts and size of proteins
were both variable then it would be
impossible to determine a proteins
molecular weight by PAGE.
4. To isolate mass as the only variable among proteins in a
sample, the strong detergent sodium dodecyl sulphate (SDS) is
added to completely unfold proteins and to normalize charge
to mass ratio.
5. The reducing agent β-mercaptoethanol is also added to
reduce disulphide bonds to help completely unfold proteins.
6. This ensure that both compact structure proteins and bulky
structure proteins migrate through the gel at a rate only
proportional to their masses and unaffected by native molecular
shape or native charge.
Oxidative Browning
Is the browning associated with cut or bruised apples,
bananas, pears, and lettuce.

The oxidation involves phenolase, also called polyphenol

oxidase (PPO). An enzyme normally compartmentalized such
that oxygen is sequestered from PPO.

Upon injury or cutting of such produce, PPO principally acts

on tyrosine residue in the presence of oxygen via an initial
hydroxylation reaction of the phenolic ring followed by the
oxidation itself.

Other phenolic compounds can also be oxidized by phenolase

to produce desireble brown pigmentation in products such as
raisins, prunes, dates, cider, and tea
There are a series of chemical reactions that begin when the apple cells are broken.
1. When the Apple sliced, these organelles, chloroplasts and vacuoles,
release polyphenol oxidase (PPO) enzyme and phenols .
2. In the presence of oxygen, the polyphenol oxidase enzymes rapidly oxidize the
phenols forming quinones. This is an aerobic oxidation reaction, which means the
phenols chemically combine with oxygen.
3. The quinones combine with each other forming a brown pigment called melanin.
As the amount of melanin increases the darker will be the surface of the sliced apple.
Proteins in Biobased Application
Next to uses proteins in the food and feed sector, proteins can
also used in more technical biobased applications. Protein rich
residues produced by the agro sector and industry, and new
protein sources, potentially can be valorised as binders in
coating and adhesives, as surface active agents and as green
Henry Ford was an early pioneer in soy protein utilization,
applying these source to improve his automobile. Products such
as plastics were developed.

Other usage of protein in bio based application are :

1. Paper Industry : sizing agents, binders, and adhesives.
2. Glue from collagen has ben used for binding paper and as an
adhesive paper coating.
3. Plywood adhesives based on soy protein have been
4. Hydrolysates from keratin, gelatin, and wheat gluten have
been used in cosmetics, as surfactant in shampo
An Alternative way of using protein is as hydrolysate. An advantage is that the
three dimensional structure of a hydrolysed protein is not present anymore and
therefore it can also be assumed that the nature of protein, denatured or not,
has less impact on the functinality of the protein hydrolysate.

Hydrolysis usually is needed to obtain protein structure with surface active

properties. Therefore Protein hydrolysates can, for example be used in cosmetics
products ( shampoo and creams) and also ceoating system.
Thank you for the attention