A.

TIT LE OF EXPERIMENT
Amino Acid and Protein

B. OBJECTIVE
Before the experiment student have to understand the structure of protein.
In this experiment, student can be expected:
1. Can prove the existence of peptide bond.
2. Can understand the reaction of Xanthoproteat and biuret test to various contents
of protein.
3. Understand the solubility and the properties of the amphoter of amino acid.

C. LITERATURE REVIEW
The relationship between structure and function reaches its ultimate
expression in the chemistry of amino acids, peptides, and proteins. Amino acids are
carboxylic acids that contain an amine function. Under certain conditions the amine
group of one molecule and the carboxyl group of a second can react, uniting the
two amino acids by an amide bond.

Amide linkages between amino acids are known as peptide bonds, and the product
of peptide bond formation between two amino acids is called a dipeptide. The
peptide chain may be extended to incorporate three amino acids in a tripeptide, four
in a tetrapeptide, and so on. Polypeptides contain many amino acid units. Proteins
are naturally occurring polypeptides that contain more than 50 amino acid units
most proteins are polymers of 100–300 amino acids (Carey, 2000: 1051).
The trivial names of the a-amino acids that are commonly found in proteins
and are represented in the genetic code, together with their symbols, systematic
names and formulas, are given in Table 1. Some other common amino acids are
listed in the. When the phrase 'amino acid' is a qualified noun it contains no hyphen;
a hyphen is inserted when it becomes an adjective so as to join its components in
qualifying another noun, e.g. amino-acid sequence (Dixon, 1984: 598).
Amino acid zwitterions are internal salts and therefore have many of the
physical properties associated with salts. They are relatively soluble in water but
insoluble in hydrocarbons and are crystalline substances with relatively high
melting points. In addition, amino acids are amphiprotic, meaning that they can
react either as acids or as bases, depending on the circumstances. In aqueous acid
solution, an amino acid zwitterion is a base that accepts a proton onto its –CO2
group to yield a cation; in aqueous base solution, the zwitterion is an acid that loses
a proton from its –NH3+ group to form an anion.

(McMurry, 2011: 504-505).

Amino acid analysis was by ion exchange chromatography (IEC) 9 using
the Technicon Sequential Multisample (TSM) Amino Acid Analyser (Technicon
Instruments Corporation, New York). The period of analysis was 76 min for each
sample. The gas flow rate was 0.50 ml min-1 at 60 0C with reproducibility
consistent within ± 3 %. The net height of each peak produced by the chart recorder
of the TSM (each representing an amino acid) was measured and calculated. The
amino acid values reported were averages of two determinations. Tryptophan was
not determined. Norleucine was the internal standard (Edeyeye, 2011: 41).
There are two acidic amino acids (amino acids with two carboxylic acid
groups): aspartate and glutamate. Aspartate is a carboxy-substituted alanine, and
glutamate has one more methylene group than aspartate. (If their carboxyl groups
are protonated, they are called aspartic acid and glutamic acid) Two amino acids
asparagine and glutamine are amides of the acidic amino acids; asparagine is the
amide of aspartate, and glutamine is the amide of glutamate. Notice that the obvious
one-letter abbreviations cannot be used for these four amino acids because A and G
are used for alanine and glycine. Instead, aspartate and glutamate are abbreviated
D and E, and asparagine and glutamine are abbreviated N and Q. There are two
basic amino acids (amino acids with two basic nitrogen-containing groups): lysine
and arginine. Lysine has an e -amino group, and arginine has a d – guanidino group.
At physiological pH, these groups are protonated. Use the e and d to remind you
how many methylene groups each amino acid has (Bruice, 2014: 1056-1057).
Proteins and peptides are amino acid polymers in which the individual
amino acids, called residues, are joined together by amide bonds, or peptide bonds.
An amino group from one residue forms an amide bond with the carboxyl of a
second residue, the amino group of the second forms an amide bond with the
carboxyl of a third, and so on. For example, alanylserine is the dipeptide that results
when an amide bond forms between the alanine carboxyl and the serine amino
group (McMurry, 2011: 511).
Protein are high molecular weight polypeptides made up of amino acids
joined by peptide bonds. Because of the presence of peptide structure and the
presence of different amino acid residues, protein react with a variety reagents to
form coloured products. Casein it is the major protein of the milk. It is a
phosphoprotein with phospate group attached to hydroxyl group of serine and
threonine residues, pH of casein is 6.6 and its isoelectric point is 4.7. This is a
noncoagulable protein an is soluble in dilute alkaline and acidic
solution (Chawla, 2014: 41).
Casein is the major protein milk, accounting for 80% of the true protein
content. Recovery of casein from skimmed milk is carried out commercially by
precipitation using either acids or rennet, giving acid casein and rennet casein,
respectively. Several acid may be used in the manufacture of acid casein, e.g.
hydrochloric, sulphuric, lactic, but hydrochloric acid is used almost exclusively.
Acid precipitation can also be carried out by using starter bacteria to develop lactic
acid. Caseion made in this way called lactein casein. The precipitation process is a
continous one in which skimmed mil first heated to about 30℃ and acidifed to Ph
4.5 by in line injection. Washing in necessary to removal residual whey solid from
the curd (Ranken, 1997: 132).
Glycine (R=H, m.p. = 240℃) is the smallest 𝛼-amino acid and has been the
object of different studies, so its stuctural properties in the gas phase are well
understood. Hydration in known to trigger the transformation of amino acid from
the neutral form observed in the gas phase to the charged zwitterion present in
condensed media. The glycine water complexes provide the simpler molecular
models of the biologically important amino acid water interaction, representing the
initial steps of the hydration process. Aspartic acid (R= -CH2-COOH, m.p. >300℃)
is a natural amino acid with a carboxyl group in the side chain. Aspartic acid follows
the trends observed by all but one of the aliphatic amino acid
studied (Rijs, 2015: 353-364).
Biuret test principle is the test is positive fro all compounds containing more
than one peptide linkage; e.g. proteins and their hydrolytic products. This test also
positive for subtances which contain two carbonyl (-CONH2) groups, joined
together or through single atom of nitrogen or carbon and similiar subtances which
contain –CSNH2, -C(NH) NH2, or –CH2NH2 groups, also respond to this test.
Xanthoproteic test principle proteoses and peptones do not form ppt. with HNO3
but their solution become yellow and then orange, when made alkaline. In this test,
a white precipitate of protein after the addition of HNO3 is due to the formation of
‘meta proteins’ which is insoluble in HNO3 . the nitro compounds from the protein
molecule containing benzene ring. These nitro compounds in alkaline medium
ionise freely and product a deep yellow or orange colour (Kulkarni, 2008: 48-49).
Xanthoproteic test the benzene ring system in tyrosine and tryptophan
undergo nitration on treatment with strong nitric acid at elevated temperatures. The
nitrated derivatives are yellow in color. When made alkaline the shades of the
colour turns to orange. On the biuret test cupric ion in alkaline medium frms a violet
coloured complex with peptide bond nitrogens of peptides and proteins. The
reaction is so named since Biuret (NH2CONHCONH2) formed by condensation of
two molecule urea, when heated at 180℃, also answer this test.
 (Chawla, 2014: 42).
Time, temperature, the presence of other subtances, and the varied nature of
amino acid chemistry are of primary importance in protein hydrolysis. Although
hydrolysis of protein is technicially as important as the analytical machinery used
to quantitate amino acids, it is usually considered secondary to the more
sophisticated analytical hardwere. However, no amount of hardwere can subtitute
for careful attention to etail during hydrolysis. The time necessary to hydrolize the
peptide bond varies with the amino acid. Generally, the peptide bonds between
aliphatic amino acids are the most difficult to break, usually 24 hr at 110 ℃ will not
fully hydrolyze these peptides (Nissen, 1992: 2).

D. APPARATUS AND CHEMICALS

1. Apparatus
a. Beacker glass 600 ml (1 piece)
b. Beacker glass 100 ml (2 pieces)
c. Test tube rack (2 pieces)
d. Funnel (1 piece)
e. Spray bottle (1 piece)
f. Round flask 50 ml (1 piece)
g. Test tube (13 pieces)
h. Graduated Cylinder 10 ml (2 pieces)
i. Watch glass (1 piece)
j. Stir bar (1 piece)
k. Spatula (1 piece)
l. Analytical balance (1 piece)
m. Drop pipette (5 pieces)
n. Reflux tools (1 set)
o. Stative and Clamp (@ 1 piece)
p. Rought Cloth (1 piece)
q. Soft Cloth (1 piece)
r. Spiritus burner (1 piece)
2. Chemicals
a. Hydrochloride acid solution 10 % and 20 % (HCl)
b. Sodium Nitrous solution 5% (NaNO2)
c. Copper (II) shulphate solution (CuSO4)
d. Aquades (H2O)
e. Nitrite acid concentrated (HNO3)
f. Glycine powder
g. Casein
h. Ice cube
i. Tyrosine solution 0,1 M (C9H11NO3)
j. Sodium hydroxide solution 10% (NaOH)
k. Litmus Paper
l. Aluminiun foil
m. Label
n. Matches
o. Tissue

E. WORK PROCEDURE
1. Solubility and Amphoteric Properties
a. Glycine was added into 2 ml of aquadest. The solution was tested the acidity
with litmus paper. The observation was repeated with L-Tyrocine
b. 1 ml of NaOH 10 % was added into 0,1 gram of L- tyrocine in 2 ml of aquadest,
and the result was noted. Litmus paper was put into the solution. Drop by drop
of acid solution was added until the solution become acid solution. The solution
was stirred during 1 minute and observed the result
c. 0,1 gram of casein was placed into test tube, 5 ml of aquadest and 2 ml of NaOH
10 % was added into the solution. Test tube was closed and shaked until formed
colloid. 2 ml of the solution was saved for the next experiment
2. Reaction with nitric acid
a. 0,1 gram of glycine was placed into test tube and 5 ml of HCl 10% was added.
The solution was cooled until 0°C in cold water. 1 ml of NaNO2 5 % was added
into solution. The result was noted
b. Casein solution that already prepared from (1c) was cooled in cold water. 1 ml
of NaNO2 solution was added into the solution. The result was noted
3. Biurette Test
a. 0,5 gram of urea was placed into test tube and heated slowly until urea was
melted and formed gas. Gas smell was noted and tested with limus paper.
Continue the heating process until gas formation was stopped and rest become
solid. Solid matter was cooled and diluted in hot water. The solution was filtered
2 ml of NaOH 10% and 3 drops of CuSO4 2 % was added. The solution was
stirred and the colour was observed. As comparison, 0,5 gram of urea was
diluted in 3 ml of water, 2 ml of NaOH 10% and 3 drops of CuSO4 2% was
added the result was compared with the result above.
b. 2 ml of aquadest and 2 drops of CuSO4 2 % was added into 2 ml of casein
solution that was prepared from (1 c). The solution was stirred and the result
was observed.
4. Xanthoproteat Test
a. 0,1 gram of casein was placed into test tube.
b. 2 ml of concentrated of nitrite acid was adde into test tube. The solution was
heated slowly.
c. Observed the colour and the solution was cooled. NaOH 10 % was added.
5. Protein Hydrolize
a. Reflux tools was arranged with round flask 100 ml. 0, 5 gram of casein was
placed into flask.
b. 20 ml of HCl 20 % was added into solution and reflux during 40 minutes. After
that solid matter was colled in room temperature and 5 ml of hydrolize result
was cooled in cold water.
c. The result 3 ml of NaOH 10 % and 2 drops CuSO4 2% solution was heated in
bunsen burner. The result was compared with the experiment above.

F. OBSERVATION RESULT
1. 1 Solubility and Amphoteric test
NO. ACTIVITY RESULT

1. Glycine crystal + 2 ml H2O → Red (Lacmus tets)

(white) (crystal) (colorless)

+
Lacmus test (red paper)

L-Tirocyne powder + 2 ml H2O → Red (lacmus

test)
(white) (colorless)
+ (white)
Lacmus test (red paper)

2. 1 ml NaOH 10 % + 0,1013 gram L-tyrocine → Colorless

(colorless) (white powder)

2 ml H2O + Lacmus paper → Blue (lacmus test)

(colorless) (red paper)

+ 10 drops of HCl 10 % + Litmus paper → Turbid, white powder

(colorless) (red paper)

 3. 0,1178 gram casein + 5 ml H2O → Turbid, white powder

(white poder) (colorless)

2 ml NaOH 10 % → Closed and shake until Formed colloid (

colloid formed colorless)

2. Reaction with nitric acid

NO. ACTIVITY RESULT

1. cooled
a. 0,1 g glycine + 5 ml HCl 10 % →

Bubble formed
(white) (colorless)

+ 1 ml NaNO2 5 %

(yellow)

cooled
b. 5 ml HCl 10 % → +1 ml NaNO2 5 %
Colorless
(colorless) (yellow)

2. cooled Colorless
Casein 1 ml → 1 ml NaNO2 5 %

(white powder) (yellow)

3. Buiret test
NO. ACTIVITY RESULT

1. 0,5018 gr Urea → Heated until melting Melted and there are

vapor, string odor while
(white crystal)
solid
And gas formed + lacmus paper (blue paper)

→ Heated
 → Cooled + Hot water

Filter + 2 ml NaOH 10 % (colorless) + 3 While turbid

drops of CuSO4 2 % → stirred
Blue → purple

As comparison:

0,5038 gr urea (white crystal) + 3 ml H2O

Colorless
(colorless) + 2 ml NaOH 10 % (colorless) +
3 drops of CuSO4 2 % Colorless

Young blue (light)

2. 2 ml casein (colorless) + 2 ml H2O Colorless

(colorless) + 3 drops of CuSO4 → stirred
Blue → Purple

4. Xanthoproteat test
NO. ACTIVITY RESULT

1. 0,1199 g casein (white powder) + Light yellow

concentrated of white acid (colorless) →
Turbid yellow
heated → cooled + NaOH 10 % → (yellow) +
base solution

5. Protein Hydrolysis
NO ACTIVITY RESULT

1. 0,5034 of casein (white powder) + 2 ml of Brown Solution

HCl 50 % (colorless) → reflux → cooled 5 ml
result experiment → cooled + NaOH 20 % +
CuSO4 (yellow) + Heated
 3 ml NaOH 10 % + CuSO4 2 %

(colorless) (2 drops yellow) Brown solution

G. DISCUSSION
Has conducted experiments with the title of amino acids and proteins, which
aims to prove the existence of a peptide bond, understand and test xanthoproteat
biuret reaction to different types of protein content, and understand the solubility
and the nature of the amphoteric amino acids. As we know the amino acids contain
two different functional groups, namely the amine group (-NH 2) and carboxyl
group (-COOH). Natural amino acids containing amine groups are attached to the
carbon-α to the carboxyl group. Meanwhile, the protein is a natural polymer formed
from amino acid units bonded to one another via peptide bonds. Therefore, the
hydrolysis of proteins will produce the amino acids, which can reach 25 kinds of
amino acids (Tim Lecturer, 2016).
1. Solubility and character amphoteric
The purpose of this experiment is to determine solubility and the nature of
the amphoteric amino acids. On testing glycine, suggesting that glycine can be
completely soluble in water. This is due to the dipolar structure. Red litmus paper
testing against glycine-making remains a litmus paper red, which means that the
solution is acidic glycine. Judging from the structure of glycine should be neutral
because it does not contain a carbonyl group and an amine group excess, but be
aware also that the isoelectric point of glycine pH of 6.0 so that the litmus indicates
the acidic nature of the fact he is neutral. His reaction:

-
H CH COOH + H2O H CH COO
+
NH2 NH3

glycine water zwitterion of glycine

 In testing the L-tyrosine, suggesting that it is not soluble in water. L-tyrosine
soluble for forming a suspension as well as the R group in L-tyrosine in the form of
an aromatic group that may cause the structure and the bond is very stable in the
presence of resonance so poorly soluble in water. This is consistent with the theory
that if the R group is composed of many carbon atoms or aromatic nature, then the
amino acid soluble in water (Tim Lecturer, 2016). While at pH testing showed L-
tyrosine acidic because when the litmus test, the suspension of L-Tyrosine used
litmus red and does not change the color of litmus. This is not in accordance with
the theory that L-tyrosine is neutral. This is due to a lack of rigor at the time of
reacting a compound. Reactions that occur are:

-
COOH COO

+
H2N C H + H2O H3N C H + H2O

CH2 CH2

OH OH

On casein test showed that he was poorly soluble in water, because the structure
containing benzene chain. In testing the pH, alkaline casein.
Furthermore, in this experiment L-tyrosine is added with 10% NaOH which
causes the pH becomes alkaline. After that drip with HCl so that it is acidic. This
shows that L-tyrosine are amphoteric because it can react with acidic or
alkaline. Reaction:

-
HO CH2 CH COOH + H2O HO CH2 CH COO + H2O
+
NH2 NH3

L-Tyrosine water zwitterion water

-
HO CH2 CH COO + NaOH HO CH2 CH COONa + H2O
+
NH3 NH2

zwitterion base Sodium tyrosine water

 HO CH2 CH COONa + HCl HO CH2 CH COOH + NaCl

NH2 NH2

Sodium tyrosine acid L-Tyrosine salt

-
HO CH2 CH COOH + HCl HO CH2 CH COOH + Cl

NH2 NH2

L-Tyrosine acid L-Tyrosine Ion Cl-

In this experiment, casein is added with water and then added with NaOH
which serves to provide alkaline conditions to the solution, formed colloid shows
that casein reacts with a base. The turbid solution is caused by the carbon chains
that bound causing poorly soluble casein. This is consistent with the theory that if
the R group is composed of many carbon atoms or aromatic nature, then the amino
acid soluble in water (Tim Lecturer, 2016). Reaction that occur:
O O

NH CH C NH CH C NH CH + H2O + NaOH

CH2 CH2 CH2

OH OH OH
n

Casein water Sodium hydroxide

2. Reaction with acid nitrites

The objecrive of this experiment is to detect the presence of amine
groups. At the time of glycine is reacted with HCl, then added again with NaNO2 to
produce a clear solution, and there are lots of gas bubbles. This shows that this
amino acid containing amine groups that react dnegan nitrite (HNO2) producing N2
gas. Reaction that occur:
 HCl + NaNO 2 HNO 2 + NaCl

H CH COOH + HNO 2 H CH COOH + H2O + N2

NH2 OH
(nitrogen gas)

While the in solution without glycine that is a mixture of HCl and NaNO2 to
produce a clear solution and does not form bubbles. This is because there are no
free amine group in HCl, as HCl not including amino acids. As for the reactions
that occur are:

NaNO2 → HCl + HNO 2 + NaCl

In this experiments casein is added with NaNO2 to give a solution without

bubbles. This shows there is no free amine groups that do not form N2 gas. His
reaction:

O O O
HN CH C NH CH C NH CH C . + NaNO 2
CH 2 CH 2 CH 2

OH HO n OH

2. Biuret test
The objective of this experiment is to test whether or not the peptide bond
which is characterized by the formation of purple (Chawla, 2014). Urea melted and
produce strong odors. This odor is caused by ammonia gases (NH 3). Urea is
alkaline which is marked by the changing color of litmus paper from red to
blue. The solution is compacted to form a white precipitate. Then reconstituted with
hot water to form a white precipitate. Then the solution is filtered to separate the
precipitate and filtrate. Filtrate obtained is then added with NaOH. Which serves to
prevent the formation of deposits of Cu(OH)2. The resulting solution is a solution
that is colorless (clear) and then make the addition of CuSO4 solution which
produces a purple solution. This proves the existence of a peptide bond. As for the
reactions that occur are:

O O O O

H2N C NH2 + H2N C NH2 H2N C N C NH2 + NH3

H

urea biuret ammonia gas

NH2 NH2

H2N C NH C NH2 + 2 NaOH + CuSO 4 O C C O + Na 2SO4

O O 2+
H N Cu N H

O C C O

NH2 NH2

senyawa kompleks

For comparison, the first urea dissolved into water, NaOH and CuSO 4 and produce
a light blue colored solution. This indicates that no peptide bond is formed, because
in this experiment is not done warming to form a peptide bond between the
molecules of urea. As for the reactions that occur are:

H2N C NH2 + NaOH + CuSO 4

urea

Further, casein is dissolved in water and added a solution of NaOH. The

addition of NaOH function is to provide the alkaline solution. Then added CuSO4 as
a provider 2+ ions will form a complex compound. The resulting solution was bright
blue. Noting there was no formation of peptide bonds. This is not in accordance
with the theory that there is a bond in the casein peptide (Chawla, 2014). This is
because the casein solution was made on the first try has long kept, and not covered
with aluminum voil so the solution contained in the tube contaminated with the
outside environment. As for the reactions that occur are:

CuSO 4. 5H2O + 2 NaOH Cu(OH) 2 + Na 2SO4 + 5 H 2O

 2+ -
Cu(OH)2 Cu + 2OH

O O

2+
H2N CH C NH CH C NH CH COOH + 2Cu

CH2 CH2 CH2

OH OH OH
n

OH OH n OH

O O
CH2 CH2 CH2

H2N CH C HN CH C NH CH COOH

2+
Cu

H2N CH C HN CH C NH CH COOH

CH2 O CH2 O CH2

OH OH OH
n

3. Test Xantroproteat
The objective of this experiment is to prove or identify the presence of
benzene or aromatic groups on amino acid which is characterized by the formation
of a yellow solution (Kulkarni, 2008). In this experiment the casein dissolved in
concentrated nitric acid and produce a yellow solution. The function of the addition
of concentrated nitric acid in this case is as a donor of NO2 - After the solution was
heated and ditambahkna with 10% NaOH produce a yellow solution, but after
adding excessive alkaline solution turns into a murky yellow color (colloidal
solution). This represents an aromatic ring on casein experiencing nitration upon
the addition of nitric acid so produce yellow nitro.
H2N CH C NH CH C NH CH COOH + (HNO 3)2

CH2 O CH2 O CH2

OH OH OH
n

H2N CH C NH CH C NH CH COOH + nH 2O

CH2 O CH2 O CH2

O 2N NO 2 O 2N NO 2 O 2N NO 2

OH OH n OH

H2N CH C NH CH C NH CH COOH + n NaOH

CH2 O CH2 O CH2

O 2N NO 2 O 2N NO 2 O 2N NO 2

OH OH n OH

H2N CH C NH CH C NH CH COOH + n H 2O

CH2 O CH2 O CH2

O 2N NO 2 O 2N NO 2 O 2N NO 2

ONa ONa n ONa

4. Protein Hydrolisis
 This experiment aims to break the peptide bond. In this experiment the
casein dissolved in HCl produce a clear solution which means a solution dissolve
completely. Extra functions HCl in this case to expedite termination or peptide bond
can be regarded as a catalyst and at reflux. The function of reflux itself is to break
peptide bonds due to the high temperature bonding this peptide will drop out due to
rupture hydrogen bonds that sustain the structure of amino acids, so that the
hydrophobicity of the side groups of polypeptides will open where the termination
is due to power withdrawal of electrons from the side and the concentration of
hydroxyl ions. Before the process reflux, prior to the round flask was added boiling
stone that serves to prevent burst upon heating. Then the result of reflux is divided
into two. The first is cooled with ice water and the second coupled with NaOH and
CuSO4 that produces brown solution. This proved to hydrolysis, peptide bonds
disconnected so that when the addition of CuSO4 produce a negative
result. Reaction that occur is
HCl
H2N CH C NH CH C NH CH COOH + NaOH + CuSO 4

CH2 O CH2 O CH2

OH OH OH

The true reaction is :

HCl
H2N CH C NH CH C NH CH COOH H2N CH COOH

CH2 O CH2 O CH2 CH2

OH OH OH OH
n
H. CONCLUSION AND SUGGESTION
1. Conclusion
From the experiment we can conclude that:
a. The presence of peptide bond is can be proof in biuret test, which the postiif test
of this test is the presence of purplish violet color.
b. Xanthoproteat reaction is protein test to prove the existence of the benzene ring
in the protein. Biuret test is a test to prove their peptide bonds in proteins.
Xanthoproteat reaction evidenced by the white blob. Biuret test proved with a
purple solution.
c. Amino acids have two functional group that different are amines group (NH2)
and carboxil group (COO-) so that asam amino can be acids and also alkaline
or we can say that amino acid is amphoteric.
2. Suggestion
As the suggestion for the next experiment is the apprentice used to be master
by reacting all the compound and also should really master the theory and the work
procedure in order that to reduce the mistaken and the result that we obtained is
better.
 BIBLIOGRAPHY

Bruice, Paula Yurkanis. 2014. Organic Chemistry Seventh Edition. California:

PEARSON.

Carey, Francis A. 2000. Organic Chemistry Fourth Edition. America: McGraw Hill
Companies, Inc.

Chawla, Ranjna. 2014. Practical Clinical Biochemistry Methods and

Interpretations Fourth Edition. India: Jaype Brothers Medical Publishers.

Dixon, H.B.F. 1984. Nomenclature and Symbolism For Amino Acids and Peptides.
Internasional Union of Pure and Applied Chemistry. Chem Vol. 56 No. 5.

Edeyeye, E.I. IG Adanlawo. 2011. Amino Acid Composition of the Ripe Fruits of
Solanum Aethiopicum and Solanum Macrocarpon. Internasional Journal of
Pharma and Bio Science. Vol 2 Issue 2.

Kulkarni, M.V. et al. 2008. Biochemistry. Pune: Nirali Prakashan.

McMurry, John. 2011. Fundamental of Organic Chemistry Seventh Edition. USA:

Cengange Learning.

Nissen, Steven. 1992. Modern Methods in Protein Nutrition and Metabolism.

California: Academic Pres, Inc.

Ranken, M.D. et al. 1997. Food Industries Manual 24th Edition. UK: Blackie
Academic and Professional.

Rijs, Anouk M. et al. 2015. Gas-Phase IR Spectroscopy and Structure of Biological

Molecules. London: Springer.
 JAWABAN PERTANYAAN

1. a. Rumus molekul glisin : CH2 – COOH

NH2
Glisin merupakan asam amino netral karena tidak memiliki gugus amino
maupun gugus karboksil pada rantai sampingnya (R) atau pada strukturnya.
b. Rumus molekulnya L- aspatrat : COOH – CH2 – CH – COOH
NH2
Asam L – aspartat merupakan asam amino asam karena memiliki gugus
karboksil pada rantai sampingnya. Gugus karbiksil ini akan melepas proton
ke dalam air sehingga terbentuk ion aspartat (suatu anion).
HOOC – CH2 – CH – COO- + -OOC – CH – COO- + H3O
+ +
NH3 NH3
Asam aspartat ion aspartat
a. Rumus molekul Tirosin : OH – CH2 – CH – COOH
NH2
Tirosin merupakan asam amino netral karena tidak memiliki gugus amin
dan karboksilat pada rantai sampingnya (R)
2. Reaksi yang dapat dijelaskan bila larutan alkali perlahan-lahan diasamkan yaitu
:
a. Larutan alkali basa
NaOH + HCl NaCl + H2O

Pada reaksi ini, larutan alkalis basa menghasilkan garam dan air.

b. Larutan alkali asam

2 KCl + H2SO4  K2SO4 + HCl
 Pada reaksi ini, larutan alkali asam menghasilkan garam dan asam.

3. Perbedaan sifat kasein dengan hasil hidrolisisnya terhadap asam nitrit dan
terhadap iji Biuret
a. Terhadap asam nitrit: terjadi hidrolisis ikatan peptida dari polimer
protein. Hidrolisis ini menghasilkan monomer asam amino dimana
ikatan peptida tidak lagi terbentuk hingga terdapat gugus amin bebas
pada kasein tersebut.
b. Terhadap uji biuret: kasein masih berada dalam bentuk protein karena
terdapat gugus peptida (COO – NH ) dan merupakan reaksi warna yang
umum untuk peptida yang ditandai dengan terbentuknya warna ungu.
Reaksi warna ini terjadi karena terbentuk senyawa kompleks antara Cu2+
dengan N dari molekul peptida.
 QUESTION
1. Glysine is an amino acid neutral because it has amino and carboxyl group on
the side chains (R) or in the structure

L-aspartic acid is an amino acid because it has carboxyl groups on the side
chains. Carboxyl group will release protons in water to form aspatate ion (an
Anion).

Tyrosine is an amino acid neutral because it has no amine groups and

carboxylate side chains (R)

2. The reaction:

3. Differences in the nature of casein with the results of the nitric acid hydrolysis
and the biuret test.
a. Against nitric acid : hydrolysis of peptide bonds of protein polymer. This
hydrolysis produces amino acid monomers in which a peptide bond is
formed until there is no longer a free amine group on the casein.
b. Agains biuret test : casein is still in the form of protein because tnere is a
cluster of peptides (COO-NH) and a color reaction that is common to the
peptide characterized by the formation of a purple color. This occurs
because the color reaction compound formed between the Cu2+ coplexes
with N of peptide molecules.
 APPROVAL SHEET

Complete report of Organic Chemistry II Experiment, which entitle “Amino

Acid and Protein” arranged by:

name : Ummu salamah

reg.number : 1513440005
class/group : Chemistry education of ICP/ IV
it has been checked well by Assistant and Assistant Coordinator and acceptable

Makassar, December 2016

Assistant Coordinator Assistant

Riswan Dwi Antoro Riswan Dwi Antoro

ID: 1313442007 ID: 1313442007

Known by,
Responsibility Lecture

Iwan Dini, S.Si, M.Si

ID.19781205 200604 1 002