Biomaterial Database

High-level secretion of human bikunin from recombinant Pichia pastoris. 2015

M Yao, and J Zhang, and X Wang

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.

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. CONCLUSIONS This study demonstrated the highest rh-bikunin production process towards its application as trypsin inhibitor in clinic. In this work, Pichia pastoris GS115 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.

UI	MeSH Term	Description	Entries
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D010843	Pichia	Yeast-like ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES isolated from exuded tree sap.	Hansenula,Hansenulas,Pichias
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002845	Chromatography	Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.	Chromatographies
D005285	Fermentation	Anaerobic degradation of GLUCOSE or other organic nutrients to gain energy in the form of ATP. End products vary depending on organisms, substrates, and enzymatic pathways. Common fermentation products include ETHANOL and LACTIC ACID.	Fermentations
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000510	Alpha-Globulins	Serum proteins that have the most rapid migration during ELECTROPHORESIS. This subgroup of globulins is divided into faster and slower alpha(1)- and alpha(2)-globulins.	Alpha-Globulin,Alpha Globulin,Alpha Globulins
D014357	Trypsin	A serine endopeptidase that is formed from TRYPSINOGEN in the pancreas. It is converted into its active form by ENTEROPEPTIDASE in the small intestine. It catalyzes hydrolysis of the carboxyl group of either arginine or lysine. EC 3.4.21.4.	Tripcellim,Trypure,beta-Trypsin,beta Trypsin
D014361	Trypsin Inhibitors	Serine proteinase inhibitors which inhibit trypsin. They may be endogenous or exogenous compounds.	Trypsin Inhibitor,Inhibitor, Trypsin,Inhibitors, Trypsin