Hansunella polymorpha is extensively used as production strain for expressing recombinant proteins including recombinant Hepatitis B surface antigens (HBsAg). HBsAg is expressed in H. polymorpha and remains bounded to the membrane. Yeast cells are having very tough cell wall because of this; there is very little effect of chemicals on yeast cells for disruption. Some enzymes act on the yeast cells to break open the content, but their application is not feasible for industrial purposes. We established the cell lysis by using the mechanical disintegrater which gives more than 85% of cell lysis in one shot. This cell lysate was subjected to different types of chaotropic agents in different concentrations to selectively leach out the membrane bound HBsAg. Each chaotropic agent showed different levels of effect during extraction. The highest protein quantity was recorded with sodium chloride and least was with urea. Further, the quality of HBsAg extracted using urea was superior in comparison with other chaotropic agents. It was also found that the enrichment of HBsAg was found to be more in potassium thiocynate. In another experiment, the cells were exposed to the non-ionic detergents (triton X-100) before subjecting to the cell lysis. We found that there is strong link between the treatments of cells with non ionic detergent and leaching of the membrane bound protein. Very less protein could be extracted from the cells which are not exposed to non-ionic detergent. This was confi rmed by running several experiments keeping the entire cell lysis parameters constant, only cells were either exposed or non exposed.
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