alexa High-level expression of non-glycosylated and active staphylokinase from Pichia pastoris.


Journal of Addiction Research & Therapy

Author(s): ApteDeshpnade A, Mandal G, Soorapaneni S, Prasad B, Kumar J,

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Abstract Staphylokinase (SAK) is a promising thrombolytic agent for treating blood-clotting disorders. Recombinant SAK (rSAK) was produced after integration of the gene into Pichia pastoris genome. The recombinant Pichia carrying multiple insertions of the SAK gene yielded high-level (approximately 1 g/l) of extracellular glycosylated rSAK (approximately 18 kDa) with negligible plasminogen activation activity. Addition of tunicamycin during the induction phase resulted in expression of non-glycosylated and highly active rSAK (approximately 15 kDa) from the same clone. Two simple steps of ion-exchange chromatography produced an homogenous rSAK of >95\% purity which suitable for future structural and functional studies. This article was published in Biotechnol Lett and referenced in Journal of Addiction Research & Therapy

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