1. use. They have the following applications: ·

1. Introduction

Proteases and their uses:      

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            Proteases or proteolytic enzymes are
enzymes that are seen to have multiple biological applications in plants and
animals, such as germination, inflammatory processes, complement activation and
many more. They can be divided into two broad subcategories, namely exotoxins and endotoxins.

            For commercial purposes, proteases
are used in leather, food and textile industries. Of both classes of proteases,
endopeptidases have much more significance in industrial use. They have the
following applications:

of chymosin in cheese preparation

sauce preparation


an alternative to chemical detergents in the leather industry


Isolation from plant sources over microbial sources:

          For industrial purposes, proteases are usually isolated
from microbial sources. But of recent, plant sources have showed promise due to
a variety of factors, such as their broad substrate specificity, allowing for
more flexible substrate preparations for varied plant sources. They also have a
more wide range of permissible temperature, pH and other factors such as the
presence of organic compounds.

            Proteases are seen to be at a high
level especially during and post germination in seeds. This is due to the fact
that these enzymes play an important role in the various biochemical mechanisms
involved in germination and also in the initial stages of development. Of plant
seeds, legume seeds have higher levels of globulin and albumin storage
proteins, which are used for the nourishment of the seedling. Due to the above
factors, the following investigation of comparing the protease content in
multiple samples has been performed.


2. Review
of Literature

2.1 In 2013, Ranajit Kumar Shaha and Shyam Sundar Shaha
studied and compared the germinating conditions and protease activity in
multiple leguminous samples including black gram, green gram, etc. They further
performed time course studies for the same and obtained an outline of when best
to perform protease iisolation for maximum yield.

2.2 In 2013, Anupama V., Marimuthu M. and others studied
and performed the partial characterization of proteases from underutilized and
common food legumes. Their method of isolation was seen to be a very simple,
inexpensive procedure for the isolation of the enzyme.

2.3 In 2016, Diego F. Coelho and Elias
Basile Tambourgi performed a study on the use of Azocasein as a substrate for
protease activity determination. Their research provided a reliable procedure
for analyzing the biological activity of proteolytic enzymes.

2.4 In 2014, M. Akhtaruzzaman and Tanjina
Rahman presented the characterization of protease enzymes from seven leguminous
seeds. Their study showed which seeds showed the highest quantity of the
proteolytic enzyme.


3. Aim and Objectives:

3.1 Aim

To compare and contrast the
protease content in green gram (Vigna radiata)
and Bengal gram (Cicer arietinum).




3.2.1 To prepare
a crude enzyme from the sample obtained through straining and precipitation

3.2.2 To estimate
the protease content in the samples using UV/Spectrophotometry

3.2.3 To compare
the protease content in the samples and identify whichever has the higher
amount present.


4. Materials and Methods

4.1 Germination

          The samples chosen for this
procedure (green gram and Bengal gram) were cleaned thoroughly. They were
surface sterilized with 70% ethanol and repeatedly washed with distilled water.
The seeds were then allowed to germinate in room temperature (28-30°C) in
cycles of 12 hours darkness and 12 hours light for 24 hours. During this phase,
they were wrapped in moistened cloth and kept in a closed vessel during the
dark phases. After 24 hours, they were used for the expermint.