I am trying to express a plasmodium enzyme (83kda) in pichia pastoris x33 strain using pGAPZ alpha A vector.

I don't see any protein secreted in the media nor in the cell fraction (western blot was done at time points 24hr, 48hr, 60hr, 72hr).

 RT-PCR shows the presence of full length transcript. Why is the translation not happening? The codon bias has been taken care of, we had found two rare codons and have mutated them.

Please suggest any way to get protein expression?