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High‐level secretion of human bikunin from recombinant Pichia pastoris
Author(s) -
Yao M.,
Zhang J.,
Wang X.
Publication year - 2015
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12470
Subject(s) - pichia pastoris , chemistry , recombinant dna , pichia , yeast , biochemistry , secretion , chromatography , gene
Human bikunin is a glycoprotein that exhibits trypsin inhibitory activity against serine proteases, and is effective in clinic. However, limited productivity and high price of human bikunin retard its further application. In this study, a high‐yield, low‐cost process of recombinant human bikunin (rh‐bikunin) production from Pichia pastoris was established. The trypsin inhibitory activity reached 6·2 × 10 3 IU ml −1 after 120 h induction of P. pastoris fermentation process, which was 20‐fold higher than that of the previous yield. Furthermore, a simple and low‐cost purification process, including ammonium sulphate precipitation, anion exchange adsorption of impurity and cation exchange chromatography, was developed with the results of 38·7% recovery and 96·6% purity of rh‐bikunin. This work made a big step to improve bikunin further application in clinic. Significance and Impact of the Study This study demonstrated the highest rh‐bikunin production process towards its application as trypsin inhibitor in clinic. In this work, Pichia pastoris GS 115 was used as a host for higher rh‐bikunin production which was 20‐fold higher than that of P. pastoris X‐33. Then, a simple, low‐cost purification procedure of rh‐bikunin was developed. This potential high productivity and low cost of rh‐bikunin process will benefit patients eventually.