Biochemical and molecular characterisation of Tetrahymena thermophila extracellular cysteine proteases

BMC Microbiology
Lutz HerrmannM Hartmann

Abstract

Over the last decades molecular biologic techniques have been developed to alter the genome and proteome of Tetrahymena thermophila thereby providing the basis for recombinant protein expression including functional human enzymes. The biotechnological potential of Tetrahymena has been proved in numerous publications, demonstrating fast growth, high biomass, fermentation in ordinary bacterial/yeast equipment, up-scalability, existence of cheap and chemical defined media. For these reasons Tetrahymena offers promising opportunities for the development of a high expression system. Yet optimised high yield strains with protease deficiency such as commonly used in yeast and bacterial systems are not available. This work presents the molecular identification of predominant proteases secreted into the medium by Tetrahymena thermophila. A one-step purification of the proteolytic enzymes is described. The information provided will allow silencing of protease activity by either knock out methods or by Tetrahymena specific antisense-ribosome-techniques. This will facilitate the next step in the advancement of this exciting organism for recombinant protein production.

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Citations

Jul 5, 2008·Journal of Cell Communication and Signaling·Morten I Rasmussen, Denys N Wheatley
Mar 30, 2010·Eukaryotic Cell·Catherine L MadingerJack S Benner
Sep 1, 2007·Fish & Shellfish Immunology·A ParamáJ Leiro
Nov 18, 2020·International Journal of Biological Macromolecules·Jihen ElleuchSlim Abdelkafi

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Datasets Mentioned

BETA
SB210
8254649
8254370

Methods Mentioned

BETA
transfection

Software Mentioned

TIGR
Multalin
SignalP

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