Survey of Digestive Enzyme in Cockroach

SURVEY OF DIGESTIVE ENZYMES IN COCKROACH
Aim: To analyse qualitatively and determine the presence of enzymes such as

invertase, amylase, protease and lipase.
Materials required: Test tubes, mortar, pestle, film strip, porcelain tiles, and pipette.
Reagents: 5% sucrose solution, Fehling’s A solution, Fehling’s B solution,

physiological saline, Benedicts saline, methyl salicylate, phenol red, 1% starch and 1%
iodine.
Procedure:
Remove the digestive tract of the anaesthetized cockroach by holding the animal in
saline and pulling off the head with attached gut. Prepare extracts of foregut, midgut and
hindgut by grinding it in a mortar. Keep the extracts in a separate test tube.
Invertase:
Take 1ml of sucrose in a test tube and add 3 drops of extract. Keep at room
temperature for one hour. Add four drops of water, 1 drop of Fehlings A and one drop of
Fehlings B in a test tube. To this add a drop of the extract and heat. A red
precipitate indicates that sucrose has been split to glucose and fructose and the enzyme
responsible for this hydrolysis of sucrose is invertase.
Amylase:
Test 1: To 1ml of 1% starch solution add 5 drops of tissue extract and shake well. From
time to time put a drop of extract in a porcelain tile and add a drop of iodine. The intensity
of blue colour will fade indicating the action of amylase. A blue colour indicates presence of
starch. Starch is initially hydrolysed to dextrins (producing reddish – violet colour) and
finally to maltose (no colour reaction). This is the achromatic point.
Test 2: To 5ml of Benedict’s solution, add 5 drops of the extract and add 5ml of starch
solution. Boil for 2 minutes and then cool slowly. Appearance of orange red colour
precipitate indicates hydrolysis of starch to monosaccharide fractions.
Protease:
Use pieces of uniformly blackened exposed and developed photographic film. This
contains a coating of gelatin. Place a single drop of extracts to be tested on the emulsion
(rough) side and let the strips rest in a moist chamber. After 30 minutes, wash off the drops
of the gelatin emulsion. The degree of digestion of gelatin by the extract is indicated by the
marks left on the photographic film.
Lipase:
To 1 ml of methyl salicylate, add a few drops of extract and add about 3 drops of
phenol red. Change in colour from pink to orange or red indicates lipase activity.
1
Result:
Foregut - Invertase, Amylase
Midgut - Protease, lipase

Hindgut - No reaction
Inference:
Animal food consists of organic material, most of which belongs to three major
groups: proteins, fats and carbohydrates. The large molecules of food are first broken down
into simpler units (monomers). These are then absorbed and either incorporated into the
body or metabolized to provide energy. The breakdown is achieved in the digestive tract
with the aid of enzymes.
Omnivorous insects like the cockroach secrete protease, lipase, amylase, invertase
hydrolyzing proteins and fats.
Digestion can take place either within or outside of cells. In a unicellular animal,
digestion is usually, of necessity, inside the cell. For example, a protozoan takes food into
the digestive vacuole, and enzymes that aid in the digestion of carbohydrates, fats, and
proteins are secreted into the vacuole. Extracellular digestion is usually associated with a
well-developed digestive tract that allows secreted enzymes to act on the food material.
The digestive tract of a cockroach is a tube modified into subdivisions, which serve
specialized digestive functions: food reception, conduction and storage, internal digestion,
absorption, conduction, and formation of feces. The three divisions of the cockroach
digestive tract are: the foregut, which includes the crop and proventriculus, midgut, which
includes the section below the proventriculus up to the caeca, and the hindgut, which
includesthe Malpighian tubules and the rectum.
The digestion starts in the salivary gland and is contained in a midgut which is
important for secreting many enzymes for digestion. The hepatic caeca of the midgut
produce an invertase, maltase, lipase and protease. The remainder of the mid-gut (stomach)
secretes the same enzymes though less abundantly. In the alimentary canal of the cockroach,
amylase is confined to the salivary glands. The amylase in the saliva of cockroach is
inactivated if it is freed from chlorides by dialysis. The amylase of cockroach with its acid
crop, has an optimum pH of 5.9.
A wide range of carbohydrates (invertase) are present commonly in insects.

Invertase, the sucrose – hydrolyzing enzyme is of common occurrence in cockroach.
Invertase breaks down complex sugars such as sucrose into simple sugars, such as fructose
and glusoce.
Proteases digests protein breaking down these large molecules into amino acids, will
digest the gelatin on the emulsion side of exposed and developed photographic film. It has
an optimum pH of 7.5 but it remains active well in the acid side of the neutrality.
2
Malphigian tubules extract could not themselves digest protein, they could facilitate
protein breakdown by the midgut secretion. Malphigian tubules are concerned with intestinal
digestion.
Unlike mammals, the salivary glands of the cockroach appear to also have lipase
activity. Lipase among invertebrates largely differs from those of vertebrates. Many
invertebrate lipases hydrolyse esters of lower fatty acids more easily than oils and fats.
Lipase accelerates the breakdown of fats into fatty acid and glycerol and continues this
breakdown action in the intestine as long as the intestine will absorb the products of
breakdown. In the tissues reversible reation takes place and during this preoess the lipse
present in the tissues convert the fatty acid and glycerol into fat. The enzyme actively is less
in foregut and digestion mainly occur in the midgut brought about by less enzymes.
The hind gut does not secrete digestive enzymes. Its main function appears to be the
elimination of waste. Enzymes may be found in the contents of that portion nearest the mid
gut, but are not demonstrable near the rectum. From this, it would appear that the
enzymesare either reabsorbed or destroyed in the hind gut.
The enzymes invertase is present in the foregut, protease and lipase are present in the
midgut, amylase is present in good quantity in the salivary gland. This shows that salivary
glands are concerned with the digestion of carbohydrates, enzymes present in the foregut,
midgut digests proteins and fats.
COCKROACH - DIGESTIVE SYSTEM
3
SURVEY OF DIGESTIVE ENZYMES IN COCKROACH
Digestive enzymes Foregut Midgut Hindgut
Amylase
Iodine test
Benedict test
Invertase
Protease
Lipase

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SURVEY OF DIGESTIVE ENZYMES IN COCKROACH

Aim: To analyse qualitatively and determine the presence of enzymes such as

Reagents: 5% sucrose solution, Fehling’s A solution, Fehling’s B solution,

Midgut - Protease, lipase

A wide range of carbohydrates (invertase) are present commonly in insects.

COCKROACH - DIGESTIVE SYSTEM

Digestive enzymes Foregut Midgut Hindgut

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