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ABSTRACT
The
goal of the proposed project is to explore/develop heat stable/protease
resistant native and immobilized wild type (from various sources) and mutant
phenylalanine ammonia- lyase (PAL) and –mutase (PAM) for biocatalysis. Based
on the considerations regarding the stability differences between eukaryotic
and prokaryotic PAL, a modified PAL construct, containing the catalytic
domain of the eukaryotic PAL and the C-terminal domain of a bacterial PAL,
will be assembled. The strategy of removing the destabilizing C-terminal
region will be used to obtain more heat and protease resistant PAM enzymes,
which are known so far only from eukaryotic sources. Successful execution of
the proposed work will provide highly active, stable enzymes with broad
substrate tolerance for the production of unnatural enantiopure (R)- and (S)-alfa- and beta-arylalanines, which are interesting building
blocks for chemical and pharmaceutical synthesis. The use of immobilized PAL
and PAM will permit the design of easily scaled-up continuous flow procedures
for industrial manufacturing of the target compounds. |