FOST107B Food Chemistry I

EXCERCISE NO.6 CHEMICAL AND PHYSICAL PROPERTIES OF

PROTEINS AND AMINO ACIDS (Experiment 2)

Abstract

Amino acids are the building blocks of all peptides and proteins and determine many of their
important properties. Protein, highly complex substance that is present in all living organisms.
Proteins are of great nutritional value and are directly involved in the chemical processes
essential for life. In this experiment we determine some of the chemical and physical properties
of proteins and amino acids and their interrelationships of the processing parameters of proteins
and amino acids. Also we aimed to study the solubility in all different types of proteins and what
are effect of temperature and pH in all types of solvent on the solubility of proteins.

Introduction

Proteins are essential nutrients for the human body. They are one of the building blocks of body
tissue and can also serve as a fuel source. These are the emoval of one water molecule and
these amino acids interact to form a peptide linkage. The foods in the following list are the most
common sources of essential amino acids the lysine is in meat, eggs, soy, black beans, quinoa,
and pumpkin seeds. Meat, fish, poultry, nuts, seeds, and whole grains contain large amounts of
histidine. Cottage cheese and wheat germ contain high quantities of threonine. The physical
and chemical properties of amino acids and proteins are amphoteric, they act as acids and
alkalies both. Each protein has a fixed value of isoelectric point (pl) at which it will move in an
electric field. Ion Binding Capacity. The proteins can form salts with both cations and anions
based on their net charge.

Proteins in foods will type complexes with alternative food elements, together with polyphenols,
that cause important changes in their structural, useful and nutritional properties (Shahidi, 2000;
Ozdal et al., 2013) (Bordenave et al., 2014; (Shahidi and Ambigaipalan, 2015). Understanding
of those changes as a results of interactions with phenolic compounds is important in light of
their scientific, industrial, and economic significance.

The determination of proteins in foods and food product has a very important biological process,
functional, and technological significance. The ways for protein estimation rely on the
measurement of a particular part or chemical group within the proteins. the elements or groups
most commonly used are nitrogen, aromatic amino acids, or bond. supported gas content
(approx. 16 pf of supermolecule weight), either supermolecule is estimated by Kjeldahl's
technique (AOAC) or ammonia can be directly determined within the digest by color-inducing
compounds like ninhydrin, indophenol, or Nessler's chemical agent.
According to (J.C. Acton 2004) Two examples of proteins in food affecting scatter are (i) acid
precipitation or heat-induced aggregation during gelation and (ii) preparation of protein powders.
As the discrete aggregates (particles) attain adequate dimensions they begin to scatter light and
give a white appearance.

Based on (Richaed J.Block 1945) The relative amino acid composition of the various food
proteins is of paramount importance and not the absolute amounts of each essential amino acid
in percent of total nitrogen. The amino acids are listed first according to their essentiality and
then according to their chemical grouping or mode of determination.

The proportion of these amino acids varies as a characteristic of a given protein, but all food
proteins with the exception of gelatin contain some of each. Amino nitrogen accounts for
approximately 16% of the weight of proteins. Amino acids are required for the synthesis of body
protein and other important nitrogen-containing compounds, such as creatine, peptide
hormones, and some neurotransmitters. Although allowances are expressed as protein,a the
biological requirement is for amino acids. USDA Agricultural Handbook Series 8 (1976-1989)

Objectives

1. To determine the type of linkage between amino acid residues.

2. To determine the typical chemical groups in protein molecules.
3. To characterize proteins and their composite amino acids.
4. To study the solubility of different types of prteins.
5. To determine the effect of temperature, pH and type of solvent on the solubility of
proteins.

Methodology

Experiment 2: Protein Solubilty

Materials:

 Buffer solutions, pH 3 to 10  CaCl2, 2%

 Egg albumin  Wheat Flour
 Soybean  Cake flour
 I2-KI indicator
 Ethanol, 70%  Waring blender
 0.1 N KOH  Mixing bowl
 IR- moisture meter  Balance test tubes

Procedure

Transfer 5 ml each of the different buffers (pH 3, 4, 5, 6, 7, 8, 9, 10) into different test
tubes containing 1 ml albumin solution, shake and observe what happens. Place 5 ml of
albumin in a test tube. Heat in a boiling water bath and observe what happens to the
protein solution at different temperatures. Place 5 ml of albumin in a test tube. Heat in a
 boiling water bath and observe what happens to the protein solution at different
temperatures.

Results and discussion

Table 6.1. Solubility off protein at different pH.

ph 3 4 5 6 7 8 9 10

Precipitation None none none none none none none none

Other none none none none none none none none

observation

There is no precipitation formed in albumin solution. Other observation none.

Table 6.2. Effects of temperature on the solubility of proteins

Temperature 40 45 55 60 65 70 80
(C)
Precipitation None None None None none none
Soy
egg None Bubbles Yes yes yes yes Yes

Other Color No White

observation no change cloudy
change

The solubility of proteins is influenced by temperature and increases with temperature between
0 and 40-80 0c. At temperatures higher than 40-80°C, the solubility of proteins is less than that
of native proteins. Heat treatment and protein denaturation causes a marked and irreversible
reduction of protein solubility.

Preparation of gluten from bread flour.

Weigh 50 g of bread or wheat flour. Place in a mixing bowl and add 30 ml water. Mix well. Rest
the dough for 10 minutes, then mix again. The mass must be slightly dry and smooth in
consistency. This is an indicator of the formation of a framework called gluten. Mold this into a
ball and immerse it in a basin of water to soften up the starchy material. Squeeze the mass until
no more whitish material is released into the water. Test the water for protein using the Biuret
teșt (see Experiment 1) and starch using iodine test. The sticky and "gummy” substance left is
called gluten. Weigh this gummy substance. Determine the moisture using the IR moisture
meter. What kind of protein goes with the wash water?
Two kinds of proteins in wheat flour is gliadin and glutenin, form in gluten. But flour also
contains many other components starch, lipids, sugars, and enzymes that contribute to the
consistency and nutritional value of bread. When you run water over dough in this activity, you
wash away most of these other substances, isolating the gluten in the dough.
Preparation of gluten from cake flour.
Weigh 50 g of cake flour and follow the same procedure as in the preparation of gluten in bread
flour. Compare the amount of gluten obtain from both flours.
Test for Starch
Transfer a drop of iodine-potassium iodide solution into a depression of a . spot plate. Add a
drop of the washings of the sample. The formation of a blue coloration indicates the presence of
starch.
Procedure
Take about 5 g of the prepared gluten. Cut the gluten into small pieces
and place in 100 ml beaker. Wash with 10 ml of 70% ethyl alcohol. Test a portion of the alcohol
washings solution for protein using the Biuret reaction. Weigh the residue left after extraction.
What is the residue left? Dissolve the residue in 0.1 N KOH and repeat the Biuret test.
Neutralize the 1 N KOH extract with acetic acid solution. Take note of the precipitate. Test the
supernatant for protein using the Biuret test.
Conclusion

Therefore, we conclude the precise amino acid content, and the sequence of those amino acids,
of a specific protein, is determined by the sequence of the bases in the gene that encodes
that protein. The chemical properties of the amino acids of proteins determine the biological
activity of the protein. As we learn about amino acids, it is important to keep in mind that one of
the more important reasons to understand amino acid structure and properties is to be able to
understand protein structure and properties. We will see that the solubility is the amount
of protein in a sample that dissolves into solution. Proteins recommended as food additives can
be partly or completely soluble or completely insoluble in water. Protein solubility is the first
functional property usually determined during development and testing of
new protein ingredients.

Review Questions

Experiment 2

1. What is an isoelectric point? Isoelectric pH?

- The isoelectric point is the pH at which a molecule carries no net electrical charge or
is electrically neutral in the statistical mean. The isoelectric point (pI, pH(I), IEP), is
the pH at which a molecule carries no net electrical charge or is electrically neutral in
the statistical mean.
2. What is denaturation? What agents bring about denaturation?
- denaturation can be brought about in various ways. Proteins are denatured by
treatment with alkaline or acid, oxidizing or reducing agents, and certain organic
solvents. The denaturing agents are those that affect the secondary and tertiary
structure without affecting the primary structure.
3. How do you classify proteins according to solubility?
 -It classified by protein based on structure function according to shape and solubility
proteins can be broadly classified into two groups, based on their shape
and solubility. The fibrous protein where in it is insoluble with water while the globular
protein is soluble in water.
4. What components were you able to separate from the gluten of flour.
- Gluten is both plastic and its able to change the shape and elastic to be able to
spring back into place. When you knead dough, you help two proteins in wheat flour,
gliadin and glutenin, form gluten.

References

Fereidoon Shahidi, Ruchira Senadheera, in Encyclopedia of Food Chemistry, 2019

K. Gupta, G. Talwar, in Encyclopedia of Food Sciences and Nutrition (Second Edition), 2003

J.C. Acton, P.L. Dawson, in Proteins in Food Processing, 2004

Richaed J.Block, in Advances in Protein Chemistry Volume 2, 1945, Pages 119-134

USDA Agricultural Handbook Series 8, in Dietary Allowances: 10th Edition (1989)