Research

Expression of yeast deubiquitination enzyme UBP1 analogues in E. coli

Anna Wojtowicz , Anna Mazurkiewicz-Pisarek , Grazyna Plucienniczak , Diana Mikiewicz-Sygula , Luiza Chojnacka , Natalia Lukasiewicz and Andrzej Plucienniczak

Institute of Biotechnology and Antibiotics, Staroscinska 5,02-516 Warsaw, Poland

author email corresponding author email

Microbial Cell Factories 2005, 4:17doi:10.1186/1475-2859-4-17

Published:	30 May 2005

Abstract

Background

It has been shown that proteins fused to ubiquitin undergo greater expression in E. coli and are easier to purify and renaturate than nonhybrid foreign proteins. However, there is no commercial source of large quantities of specific deubiquitinating proteases. This is the reason why hybrid proteins containing ubiquitin at their N-end cannot be used in large scale biotechnological processes.

Results and Conclusion

We have described the synthesis of the yeast deubiquination enzyme UBP1 muteins in E. coli. We have shown that an efficient overproduction of the enzyme in E. coli may be achieved after the introduction of several changes in the nucleotide sequence encoding UBP1. One of the conditions of an effective synthesis of the UBP1 muteins is the removal of the 5'-end sequence encoding the transmembrane region of the enzyme. The obtained variants of the enzyme may be successfully used for processing large amounts of hybrid proteins comprising ubiquitin or tagged ubiquitin at their N-ends.