Amplification of leader proregions as a mean to increase the secretion of antibody fragments in the Pichia pastoris yeast

The dependence of secretion efficiency in Pichia pastoris cells on the copy number of proregions in leader polypeptides has been studied. The humanized light kappa-chain of the murine H3-1 antibody was used as a reporter protein. The leader pre-pro-polypeptides were composed of the signal peptide (preregion) from the α-factor precursors of Saccharomyces cerevisiae and a variable number of proregions from the prepro-precursors of the α-factor or the Hsp150p protein of S. cerevisiae or Hsp150p of P. pastoris. An increase in the proregion copy number either resulted in an almost 1.5-fold increase or a fivefold decrease in secretion depending on the proregion used. It was concluded that the enhancement of the proregion copy number could be of potential value for the intensification of protein secretion in P. pastoris.