BioChem - DOS

Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli

Microbial cell factories

Wojtowicz-Krawiec, Anna; Sokolowska, Iwona; Smorawinska, Maria; Chojnacka-Puchta, Luiza; Mikiewicz, Diana; Lukasiewicz, Natalia; Marciniak-Rusek, Alina; Wolinowska, Renata; Bierczynska-Krzysik, Anna; Porebska, Anna Joanna; Kuthan-Styczen, Jolanta; Gurba, Lidia; Borowicz, Piotr; Mazurkiewicz, Anna; Plucienniczak, Grazyna; Plucienniczak, Andrzej

<P><B>Background</B></P><P>Numerous bacterial human growth hormone (hGH) expression methods under conventional fermentation and induction conditions have been described. Despite significant progress made in this area over the past several years, production of recombinant hGH by using cellular expression systems still requires further optimization. Fusion of the ubiquitin (Ub) tag to the hGH protein allowed to increase of the overall efficiency of the biosynthesis and improve the protein stability. Ub is a protein composed of 76 amino acid residues with a molecular mass of 8.6&nbsp;kDa, expressed in all eukaryotes. This protein is an element of the universal protein modification system, which does not occur in bacteria, and is a useful carrier for heterologous proteins obtained through expression in <I>Escherichia coli.</I> Purification of Ub-fusion proteins is easier than that of unconjugated recombinant proteins, and Ub can be removed by deubiquitinating proteases (DUBs or UBPs).</P><P><B>Results and Conclusion</B></P><P>In the present study the UBPD2C protease, a stable UBP1 analog, was produced as a recombinant protein in <I>E. coli</I> and used for production of recombinant human growth hormone (rhGH). hGH was expressed as a fusion protein with Ub as a tag. Our findings show that the UBPD2C protease is very effective in removing the Ub moiety from recombinant Ub-fused hGH. The described approach enables obtaining a considerable yield of rhGH in a purity required for pharmaceutical products.</P>

2014

BioMed Central

cc-by

1475-2859

13

pp.113

10.1186/s12934-014-0113-4

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART71802975

https://microbialcellfactories.biomedcentral.com/counter/pdf/10.1186/s12934-014-0113-4

Recombinant human growth hormone; Ubiquitin; Deubiquitinating protease; Protein expression