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 (~1 g/l) of extracellular glycosylated rSAK (~18 kDa) with negligible plasminogen activation activity. Addition of tunicamycin during the induction phase resulted in expression of non-glycosylated and highly active rSAK (~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.
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We sincerely thank Dr. Kamal Sharma, Managing director, Lupin Ltd. for his constant support and encouragement.
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Apte-Deshpnade, A., Mandal, G., Soorapaneni, S. et al. High-level expression of non-glycosylated and active staphylokinase from Pichia pastoris . Biotechnol Lett 31, 811–817 (2009). https://doi.org/10.1007/s10529-009-9938-z
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DOI: https://doi.org/10.1007/s10529-009-9938-z