UNIVERSITY OF DAR ES SALAAM

COLLEGE OF NATURAL AND APPLIED SCIENCE

DEPARTMENT OF MOLECULAR BIOLOGY AND BIOTECHNOLOGY

STUDENT NAME: ERASTO, FABIAN

REGISTRATION NUMBER: 2017-04-11169

DEGREE: BSC. IN MOLECULAR BIOLOGY AND BIOTECHNOLOGY

COURSE NAME: PRACTICALS IN BIOCHEMISTRY

COURSE CODE: BN 240

COURSE INSTRUCTOR: DR. ROSE MASALU

TITLE: ISOLATION OF AMYLOSE AND AMYLOPECTIN FROM POTATO STARCH

DATE OF SUBMISSION: 24TH APRIL, 2019

ABSTRACT

The aim of this experiment is to separate the two fractions of starch, amylose and
amylopectin. It is well known that unprocessed potato starch granules are formed of a matrix of
amylopectin throughout amylose is dispersed. Under mild basic conditions, the amylose leaches
out leaving the amylopectin network unaltered. Potato starch dispersed in 0.16M NaOH and
0.6M HCL, dissolved amylose precipitated with 1-butanol. Then iodine test performed by adding
two drops of iodine solution in amylose and amylopectin(pellet) isolated.
INTRODUCTION

Starch is a polymer of glucose and commonly produced in plants as an energy and carbon
storage medium, and large volumes of starch are consumed by man for food. Starch is actually a
mixture of two different, but related polysaccharides, amylose and amylopectin. Structurally,
amylose is a linear homopolymer consisting of D-anhydro glucopyranose units linked by α-
(1→4) – glycosidic bonds, while amylopectin is a branched polymer of glucose with α-(1→6)
branch points. In contrast to cellulose, whose β-(1→4)-glycosidic bonds yield a polymer with
strong crystalline regions, the α-(1→4) bonds of amylose yield a polymer chain that is less
extended and with weaker crystalline regions, so that amylose is dispersible in cold water and
soluble in hot water (Emmett M. Partainiii, 2000). The ratio of amylose to amylopectin varies
depending on the botanical source of the starch, but typical starches are about 25% amylose. In
aqueous solution, amylose behaves either as a random coil or as a helix, and the familiar blue
color seen in starch treated with iodine is caused by the formation of an inclusion complex of
iodine and amylose. A variety of inclusion complexes of lipids, polar organic solvents, and other
hydrophobic materials can be prepared with amylose, and a recent paper describes the cross-
linking of amylose and cationic hydrophobic HEC through a helical clathrate of amylose and the
hydrophobic group.

Starch is the major component of potato dry matter and consists of amylose and
amylopectin. The structural characteristics and amylose-to-amylopectin ratio of
potato starch vary among cultivars. The nutritional and processing quality of potatoes and potato
products (frozen and dry) are greatly affected by their starch characteristics. Several chemical,
physical, and enzymatic modifications are performed to improve the processing performance of
potato starch. Most of these modifications are listed as generally recognized as safe by the safety
authorities. Several modified potato starches with slow digestibility are being developed that
may provide nutritional benefits for humans. These starches have the potential to be used for the
treatment of certain medical conditions (e.g., glycogen storage disease and diabetes mellitus.)
METHODOLOGY

In the isolation of amylose and amylopectin from potato starch the following procedures was
done includes;-

Potato was grinded and 40% aqueous slurry of potato prepared by using distilled water in
a conical flask. Then 20ml of 40% aqueous slurry of potato mixed with 60ml of NaOH swirled
gently and leaved in 5 minutes for suspension separation.

After 5 minutes, 20ml of 5% NaCL added in 0.6M HCL and mixed gently and
centrifuged at 10,000rpm for 15 minutes then supernatant was seeped in different conical flask,
pellet washed with 20ml of 1% of NaCL overnight. Then centrifuged at 10,000 revolutions per
minute (rpm) for 15 minutes, supernatant discarded.

Amylose precipitated with 1-butanol from the original supernatant that separated in the
different conical flask and stands for overnight the same as amylopectin pellet. Then amylose
collected after centrifugation at 5,000 rpm for 15 minutes. Then Two drops of iodine solution
added to amylose and amylopectin respectively.
RESULT

During isolation of amylose and amylopectin from potato starch the following result was
observed;

Table No.1 Show colour change after addition of 2 -3 drops of Iodine solution in amylose
and amylopectin pellet in eppendorf tubes.

Time Iodine Test

Amylose Colour Amylopectin Colour
0-1 minutes Blue black Reddish brown
2-3 minutes Pale Blue black Pale Reddish brown
4 minutes Retain original sample colour Retain original sample colour

Picture A: Amylopectin- Reddish brown Picture B: Amylose – Blue black

DISCUSSION

Amylose and amylopectin isolated from potato when dissolved with Iodine solution it
show blue black and reddish brown that indicates positive result in a few minutes which cannot
stand all the times that is because only two drops added in the mixture therefore it require more
amounts of Iodine solution to be dissolved in the amylose and amylopectin to show a clear
positive blue black and reddish brown.
CONCLUSION

Starch is a highly hydrophilic polymer that consists of linear amylose and highly
branched amylopectin. Starch can be easily isolated using environmentally friendly processing
technologies like sedimentation phase done in isolation of amylose and amylopectin from potato
starch in this experiment.
REFERENCE

Aspreet Singh, Lovedeep Kaur, (2016) Advances in Potato Chemistry and Technology.

Emmett M. Partainiii, (2000) In Applied Polymer Science: 21st Century,

Yasmin Y. Talib,Meena S. Karve,Shobhana V. Bhide &N. R. Kale (2006). A Simple

Preparative Method for the Isolation of Amylose and Amylopectin from Potato Starch.
LABORATORY QUESTIONS

1. Sodium Hydroxide and 1-butanol used in the experiment due to the following reasons;-
i. To reduce rapid swelling of starch at the room temperature (NaOH).
ii. For amylose precipitation (1-butanol)
iii. Separation of amylose from potato (NaOH)

2. The difference between amylose and amylopectin are the following;

NO. Amyolse Amylopectin
1. It constitutes about 20% of the starch It constitutes about 80% of the starch
2. Amylose is straight chain polymer of Amylopectin is a branched chain polymer of D-
D-glucose units glucose units
3. The bonding involved is α-1->4 The bonding involved is α-1->4 glycosidic
glycosidic linkage linkage and α-1->6 glycosidic linkage at branch
points
4. With iodine amylose stains blue With iodine amylopectin stains reddish brown
5. α and β amylases can hydrolyse α and β amylases can hydrolyse α-1->4
amylose glycosidic linkage but cannot hydrolyse α-1->6
glycosidic linkage at branch points of
amylopectin
6. More soluble in water Less soluble in water
7. Soluble in hot water without swelling Soluble in hot water with swelling
8. Soluble in hot water but does not form Soluble in hot water but form starch gel or paste
starch gel or paste

3. The following carbohydrate are not polysaccharide;-

i. Isomaltose