Abstract
The glycoprotein rBm86 has been intracellularly expressed in the methylotrophic yeast Pichia pastoris. A bench-scale study and the scale-up of the wet-milling disruption of cells after an 80-h fermentation in methanol is reported. Retention times combined with different numbers of passes were used and nonsignificant differences in degree of disintegration were found above 6 min total retention time. In addition, identical degrees of disintegration were observed in combinations adding up to 4 or 6 min retention time. An energy balance yielded a clear correlation between generated heat and retention time which allowed estimating the outlet temperature of the product. Furthermore, it was found that an interaction between the rBm86 protein and cell membranes offers protection and minimizes loss of the protein during the process.
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