- Monosaccharides

Monosaccharides cannot be hydrolyzed further into simpler form of carbohydrates.

Monosaccharides are easily absorbed in intestines. All other types of carbohydrates like
disaccharides and polysaccharides are not absorbed directly. All disaccharides and
polysaccharides are ultimately converted to monosaccharides. Monosaccharides are
important in the body especially glucose, fructose and galactose. Glucose is the most
important carbohydrate in human body(Paulsen H et al, 1968).Glucose is formed from the
hydrolysis of complex carbohydrates including starch, dextrin. Glucose is found in the blood
and provides energy to the body. Glucose is also formed from breakdown of glycogen in the
body. Fructose is a reducing sugar and forms osazone crystals. Fructose is found in fruit and
is also found in honey. Fructose can be obtained in the body by action of sucrase on sucrose.
Galactose is also a reducing sugar and forms rod shaped crystals

- Disaccharides

Disaccharides consist of two sugars that are linked by a glycosidic linkage. This glycosidic
linkage is formed by a condensation reaction that takes place between the two sugars units,
resulting in the loss of a hydrogen atom from one monosaccharide and a hydroxyl group
from the other. Disaccharides are broken down into two monosaccharide, in the small
intestine during the process of digestion

- Polysaccharides

Polysaccharides cannot be directly utilized by the body. They must first be broken down into
monosaccharide, the only sugar form the body can use. Polysaccharides contain up to 60,000
simple carbohydrate molecules. Polysaccharides are polymeric carbohydrate structures,
formed of repeating units (either mono- or disaccharides) joined together by glycosidic
bonds. Unlike other saccharides, polysaccharides tend to not have a sweet taste. Some
examples of polysaccharides include starch, cellulose and glycogen). ((Carbohydrates

1H. M. Asif, 2Muhammad Akram, 3Tariq Saeed, 2M. Ibrahim Khan, 1Naveed Akhtar,
1Riaz ur Rehman, 1S. M. Ali Shah, 1Khalil Ahmed, 1Ghazala Shaheen,

1The Islamia University of Bahawalpur. 2Shifa ul Mulk Memorial Hospital, Hamdard

University, Karachi, Pakistan. 3University College of Pharmacy, Punjab University,
Lahore.Accepted 21 January 2011)
 - Starch

Starch is a polymer of -D-glucose. It consists of two main components: a linear compound

called amylose and a branched compound called amylopectin. Amylose is a linear isotactic
polymer that typically consists of up to 3000 monomer units of glucose molecules
interconnected by -1, 4 glycosidic bonds with virtually no 1,6 Amylose generally tends to
form a relatively stiff parallel left handed single helix or a s.tiffer left-handed double helix.
The oxygen atoms in positions 2 and 6 of the glucose monomer lie on the outside surface of
the helix, with the ring oxygen pointing inward. Under some conditions, perfectly aligned
chains may form double stranded crystallites that are resistant to amylases. The second and
most abundant component of starch, amylopectin, consists of a more highly branched
polycarbohydrate with 1, 4 bonds that serve as the backbone and 1, 6 branching points
which occur approximately every 20-25 glucose units.

The structurally important helices formed by amylose may assemble into crystalline
structures called type V. This helix is composed of 6 glucose residues per turn and has been
found to have a pitch between 0.791 and 0.817 nm. While it is formed easily by the inclusion
of small molecules such as lipids into the helix cavity, the size of this cavity may vary and
has been reported to be between 1.5 and 2.9 nm depending on the method and the molecular
weight of the sample.

Many types of enzymes hydrolyze the 1,4 glycosidic bonds in starch but the most common
enzyme is called amylase and was used in this study. Starches with high amylose content
generally have low digestion rates but Rao found that rice varieties with high amylose content
had faster digestion rates compared to rice varieties with low amylose content. Crystallinity
also affects the enzyme hydrolysis with B-type crystals being less susceptible to enzyme
attack than A-type. This emphasizes not only the complexity of factors influencing digestion
properties but also the sensitivity of the kinetics of digestion to the starch chain morphology.

In the enzymatic biodegradation of starch, the enzyme first diffuses and binds to the solid
substrate, cleaves the glycosidic bonds and unbinds. The -amylase enzymatic hydrolysis is
carried out by a side-by-side digestion mechanism: the amylase binds the substrate from the

side, that is, it binds the starch molecules in a direction parallel to the strands. The reaction
rate is mainly controlled by the structure of the substrate as it affects the enzyme diffusion,
binding, and bond cleavage. Thus, the kinetics of digestion offers a good indication that
structural changes occur in starch molecules. These changes are important contributors to the
possible applications of starch polymers.

Apart from its obvious use as a food source and as an enhancer of the viscosity and texture of
food products starch has a wide range of applications. It is used as adhesives, stabilizers,
coatings, binders, and molecular sieve, an encapsulating agent for the pesticides, as well as,
in the pharmaceutical industry for coating, dusting tablets, as well as, a matrix for controlled
release drugs.

Starch or amylum is a polysaccharide carbohydrate consisting of a large number of glucose

units joined together by glycosidic bonds. It comprises of amylose and amylopectin as it
macromolecules. Starch is produced by all green plants as an energy store and is a important
energy source for humans. It is found in potatoes, wheat, rice and other foods, and it varies in
appearance, depending on its source. In the unmodified form, starches have limited use in the
food industry. In general, native starches produce weak-bodied, cohesive, rubbery pastes
when heated and undesirable gels when the pastes are cooled (Adzahan, 2002). That is why,
the food manufacturers generally prefer starches with better behavioral characteristic than
those provided by native starches. The properties of starches can be improved by various
modifications. (Journal of Agricultural Science Vol. 2, No. 2, June 2010 Modified
Starches and Their Usages in Selected Food Products: A Review Study K.A. Abbas)

Starch chain structure is the most important factor in any application [20]. An understanding
and fine control of the structure is necessary for the development of a greater range of
applications, possibly including building materials. (Volume 2, Issue 1, 2008 Structural
Changes Associated with Interactions Between Starch and Particles of TiO2 or ZnSe
Paul Bernazzani) Starch readily reacts with iodine, resulting in a very dark grey to
black stain.

Starch is a storage form of glucose in the body. Starch is made up of amylose and
amylopectin. Starch contains amylase (10-20%) and amylopectin (80-90%). Starch gives blue
colour with iodine solution. In starch linkage between glucose residues is 1-4 and at branch
point linkage is of 1-6. (Carbohydrates

1H. M. Asif, 2Muhammad Akram, 3Tariq Saeed, 2M. Ibrahim Khan, 1Naveed Akhtar, 1Riaz
ur Rehman, 1S. M. Ali Shah, 1Khalil Ahmed, 1Ghazala Shaheen,
1The Islamia University of Bahawalpur. 2Shifa ul Mulk Memorial Hospital, Hamdard
University, Karachi, Pakistan. 3University College of Pharmacy, Punjab University,
Lahore.Accepted 21 January 2011 )

The enzyme amylase is secreted out of the cells (an exoenzyme) into the surrounding media,
catalyzing the breakdown of starch into smaller sugars which can then be absorbed by the
cells for use.

Iodine reacts with starch, producing a deep purple color. As starch is catabolized and
converted to sugars, there will be less and less starch to react with the iodine. Strong amylase
producers may convert all of the starch in the agar to sugars, while weak amylase producers
may convert the starch surrounding the growth areas only. (Fall 2011 - Jackie Reynolds,
Richland College, BIOL 2421)

c. Barfoed Test

Reducing monosaccharides

Reactions:

www.biosci.ohiou.edu/introbioslab/Bios170/170_2/benedict.htm

Reducing monosaccharides are oxidized by the copper ion in solution to form a carboxylic
acid and a reddish precipitate of copper (I) oxide within three minutes. Reducing
disaccharides undergo the same reaction, but do so at a slower rate.
(www.biosci.ohiou.edu/introbioslab/Bios170/170_2/benedict.htm)

the purpose is to see if the microbe can use starch, acomplex carbohydrate made from
glucose, as a source of carbon and energy for growth. Use ofstarchis accomplished by an
enzyme calledalpha-amylase.

how isalpha-amylaseactivity determined?

A medium containingstarchis used. After inoculation and overnight incubation,iodine

reagentis added to detect the presence of starch.Iodine reagentcomplexes withstarchto form a
blue-black color in the culture medium. Clear halos surrounding colonies is indicative of their
ability to digest the starch in the medium due to the presence ofalpha-amylase.

d. Benedict's Test for Reducing Sugars

The Benedict's test allows us to detect the presence of reducing sugars (sugars with a free
aldehyde or ketone group). All monosaccharides are reducing sugars; they all have a free
reactive carbonyl group. Some disaccharides have exposed carbonyl groups and are also
reducing sugars. Other disaccharides such as sucrose are non-reducing sugars and will not
react with Benedict's solution. Starches are also non-reducing sugars. The copper sulfate
(CuSO4) present in Benedict's solution reacts with electrons from the aldehyde or ketone
group of the reducing sugar to form cuprous oxide (Cu2O), a red-brown precipitate. CuSO4
Cu++ + SO4--

2 Cu++ + Reducing Sugar Cu+ (electron donor) Cu+ Cu2O (precipitate)

The final color of the solution depends on how much of this precipitate was formed, and
therefore the color gives an indication of how much reducing sugar was present.