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ABSTRACT
Glycogen is the principal storage form of carbohydrate in the mammalian body, which is mainly present
in liver and muscles. This experiment is composed of three parts: extraction, characterization, and
hydrolysis of glycogen. Extraction was done by heat denaturation of glycogen from chicken liver and
addition of 0.1% acetic acid to improve precipitation. After extraction, 95% ethanol was used to precipitate
and purify glycogen, which was seen as white precipitate. The second part involves the general tests for
glycogen which includes Molischs Test and I2 Reaction. Positive results were seen on Molischs Test and
negative in I2 Reaction which are used to test for carbohydrates and starch respectively. The glycogen
extract produced purple interface in the Molischs Test. For the I 2 reaction, a deep red color was expected,
but there was no color change in the solution before, during and after heating. The last part of the
experiment is hydrolysis of glycogen which is subdivided into acidic and enzymatic hydrolysis.
Concentrated HCl was used in acidic hydrolysis while saliva was used in enzymatic hydrolysis. The acidic
and enzymatic hydrolysates were subjected into Benedicts Test and yielded negative results, which
indicates that glycogen is a not a reducing sugar.
INTRODUCTION
Like other polysaccharides, glycogencan also
Glycogen is synthesized from glucose by the undergo hydrolysis. During the reaction, the
pathway of glycogenesis, which occurs mainly in glucose monomer units of glycogen are separated.
liver and muscle. It is the major glucose storage This is being done by the introduction of water in
polymer in animals. It has a highly-branched the glycogen molecule with the presence of strong
structure with linear chains connected by (1-4) acid or base which is summarized in Figure 2 or it
glycosidic bonds and branched points (1-6) every can also be due to the presence of enzymes. [3]
1 in 10 glucose units. It allows the immediate Amylase, an enzyme present in saliva,
release of glucose. [1] catalyzes the hydrolysis of the glycosidic linkages
in starch.
METHODOLOGY C. I2 Reaction
Few drops of 0.01 M I 2 was added into the
I. Extraction of Glycogen from Chicken Liver glycogen extract. The mixture was warmed in a
water bath and cooled after. The color of the
An amount of 13 g of chicken liver was solution was observed before, during, and after
homogenized by using a blender. Boiling water heating of the mixture.
approximately 50 mL was poured into the
homogenized chicken liver. To precipitate the III. Hydrolysis of Glycogen
proteins, the mixture was heated in boiling water
bath for 30 minutes. To improve the precipitation, 1 A. Acid Hydrolysis
mL of 0.1% acetic acid was added. The mixture
was filtered and glycogen extract was obtained, In a test tube, 5 mL of glycogen extract, and 5
which will be used throughout the experiment. drops of conc. HCl was added. The mixture was
covered with marble and boiled in a water bath for
30 minutes. The acid hydrolysate was put in a
refrigerator for Benedicts Test on the next meeting.
B. Enzymatic Hydrolysis
Collection of Saliva
C12H22O11+H+/H2O-------->2(C6H12O6)
ch.html
2+ -1 -1
R-CHO + 2Cu +5OH --------> R-COO +Cu2O +
3H2O
[1] What Is Glycogen? (2016, August 09). Retrieved April 30, 2017, from
https://www.musclesound.com/what-is-glycogen/
[2] BBC - GCSE Bitesize: What are enzymes? (n.d.). Retrieved April 30, 2017, from
http://www.bbc.co.uk/schools/gcsebitesize/science/add_aqa_pre_2011/enzymes/enzymes1.shtml
[3] Glycogen Metabolism (n.d.). Retrieved April 30, 2017, from
http://oregonstate.edu/instruct/bb450/summer09/lecture/glycogennotes.html
[4] Marini, I. (2006, November 03). Discovering an accessible enzyme: Salivary amylase : Prima digestio
fit in ore: A didactic approach for high school students. Retrieved April 30, 2017, from
http://onlinelibrary.wiley.com/doi/10.1002/bmb.2005.494033022439/full