Mechanical disruption of Pichia pastoris yeast to recover the recombinant glycoprotein Bm86

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 a...

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Bibliographic Details
Published in:Enzyme and microbial technology 1998-07, Vol.23 (1), p.58-63
Main Authors: Canales, Mario, Buxadó, José A., Heynngnezz, Lázaro, Enrı́quez, Antonio
Format: Article
Language:English
Subjects:
bead mill
Biological and medical sciences
Biotechnology
Cell disintegration
Fundamental and applied biological sciences. Psychology
Methods. Procedures. Technologies
Others
Pichia pastoris
rBm86 protein
Various methods and equipments
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Summary: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.
ISSN:0141-0229
1879-0909
DOI:10.1016/S0141-0229(98)00012-X