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Sambhaji Chavan
PRAJ Industries Ltd
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INTRODUCTION:
Enzymes were first divided into two classes
unorganised and organized ferments.
The action of organized ferments was
thought to be due to vital activity. This
view was encouraged by the rediscovery of
bacteria by Pasture and Koch1. In 1897
Eduard Buchner discovered that yeast
extracts could ferment sugar to alcohol,
proving that fermentation was promoted by
molecules that continue to function
removed from cells. Frederick Kuhne
called these molecules ENZYMES.
Enzymes promote and control the
conversion of complex carbohydrates, fats
and proteins of our body into simple
substance, which the intestines can absorb,
and also the various reactions by which
these simple substances are used in the
body for building up new tissues or
producing energy. Enzymes are not broken
JADD, May June 2014, Vol 1, Issue 3
Sambhaji Chavan et al
ASSAY OF ENZYME:-
FOR
Partial
Purification
of
Alpha
Amylase:
Ammonium Sulphate Precipitation
The crude enzyme extract was subjected
to ammonium sulphate precipitation29.
Sambhaji Chavan et al
%Relative Activity
4.65
27.97
45.54
75.34
94.50
94.95
99.78
99.16
99.48
99.93
99.01
Sambhaji Chavan et al
Activity (U/ml)*
21.73
39.51
48
58
65.3
63.2
48.07
26.99
% Relative Activity
32.99
59.99
72.89
88.07
99.16
95.97
72.99
40.98
Sambhaji Chavan et al
Activity (U/ml)*
31.2
42.8
57.2
65.2
62.1
55.3
48.2
36.2
26.3
22.1
12.3
% Relative Activity
48.83
66.99
89.52
102.04
97.19
86.55
75.44
56.66
41.16
34.59
19.25
Sr.No
Concentratio
n (mM)
% Relative activity
Mg++
1
2
3
1
78
5
101
10
78
*
Average value of three determinations.
JADD, May June 2014, Vol 1, Issue 3
Ca++
86
93
113
EDTA
64
48
17
Sambhaji Chavan et al
Optimum Temperature
Table 3 and Figure 3 Illustrate the
optimum temperature for Alpha amylase,
as the temperature increased from 400C to
55C
enzyme
activity
increased
progressively.
Above 55C enzyme
activity decreased gradually upto 90C.
and at 120C enzyme was totally
denaturated.
Thus
the
optimum
temperature for activity was found to be
55C
Effect of metal
compounds
ions
and
other
Sambhaji Chavan et al
Sambhaji Chavan et al