Abstract
The main goal of this work is the production of recombinant biologically active
peptides fused with a Carbohydrate Binding Module (CBM). Aiming at the
optimization of large scale expression, CBM peptide production was done by
cloning CBM coding sequence in two different systems of Pichia pastoris:
pGAPZαC which has a constitutive promoter and pPICZαC which has an
inductive promoter. The integration of the CBM coding sequence in the yeast
genome was confirmed by slot-blot for both expression systems. Transcription
was analysed by northern-blot and SDS-PAGE. The results obtained with these
two expression systems were different. Apparently, there were no clones of P.
pastoris transformed with pGAPZαC-CBM that had produced any protein with
starch affinity, under the batch and fed-batch conditions tested in this work. On
the other hand, only one studied clone of P. pastoris transformed with
pPICZαC-CBM vector had produced, in batch conditions, a protein with affinity
for starch. However, under fed-batch conditions, the results obtained with this
clone were not conclusive, suggesting that conditions for large scale production
must be optimized.
Publication Type: Abstracts in Conference Proceedings