Purification and identification of recombinant hirudin and its degradation products expressed in Pichia pastoris.
The purification and identification of recombinant hirudin (r-hirudin) (rHV2-Lys47) and its several C-terminal proteolytic degradation derivatives, produced by Pichia pastoris, were described. The high-purity rHV2-Lys47 of above 99% and its three degradation products were obtained by a straightforward two-step chromatography procedure, a combination of cation exchange and reverse phase chromatography, with a recovery yield of 74% for hirudin. The purified rHV2 had the predicted N-terminal amino acid sequence and the derivatives were the degradation products of hirudin, short of one to three amino acid residues at C-terminal.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, P.R. China.
Wei-Bin
WB
Zhou
Yuan-Xing
YX
Zhang
D002852:Chromatography, Ion Exchange; D015870:Gene Expression; D006629:Hirudins / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
D006629:Hirudins; D011994:Recombinant Proteins; C064761:hirudin, Lys(47)-