Biotechnological Approaches for the Downstream Processing of Selected Enzymes

The present study mainly covers the downstream processing of selected
enzymes using reverse micellar extraction, aqueous two phase extraction and
membrane processes. Experiments were carried out to evaluate the effect of
various process parameters of these processes on recovery and purification
of target enzyme from natural sources which are often complex mixture.
Reverse micellar extraction, aqueous two phase extraction and membrane
processes are operated individually as well as in an integrated mode for the
purification of selected enzymes such as bromelain, β-galactosidase and β-
glucosidase. Major emphasis is given to develop a systematic approach for
the extraction and purification of enzymes from natural systems. Reverse
micellar extraction of β-galactosidase and β-glucosidase has given a new
insight into successful extraction of large molecular weight enzymes when
attempted by injection mode. Similarly mixed reversed micellar system is used
for the first time for extraction and purification of enzyme from real system. A
new approach for simultaneous fractionation and purification of mixture of β-
glycosidases namely, β-galactosidase and β-glucosidase, from Hordeum
vulgare using aqueous two phase system was provided. The conventional
methods of enzyme purification were also carried out and compared with the
present methods.
The present protocols, although studied for particular enzymes, can be
applied in general to any other biomolecules. The thesis contributes to applied
science in terms of process development for the extraction, isolation,
purification and concentration selected enzymes from natural complex
systems.