Efficient bacterial expression of fusion proteins and their selective processing by a recombinant Kex-1 protease

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• 作者：Sabrina Pozzuolo ; Umberto Breme ; Barbara Salis ; Geoffrey Taylor ; Giancarlo Tonon ; Gaetano Orsini
• 关键词：Fusion proteins ; Kex-1 protease ; Enzymatic cleavage ; Protein expression
• 刊名：Protein Expression and Purification
• 出版年：2008
• 出版时间：June 2008
• 年：2008
• 卷：59
• 期：2
• 页码：334-341
• 全文大小：474 K

文摘

A secreted, soluble variant of the Kex-1 endopeptidase from Kluyveromyces lactis has been produced and studied as a novel cleavage enzyme exhibiting high specificity for the Lys-Arg peptide. This highly selective, efficient enzyme is particularly adapted for use in manufacturing when a recombinant therapeutic protein, possessing its native N-terminus, has to be released in vitro from a bacterially-expressed fusion protein. In this paper, we describe the preparation of a Kex-1 variant using Saccharomyces cerevisiae and its application in the production of important therapeutic recombinant proteins such as human growth hormone, granulocyte colony-stimulating factor and interferon--2b.