Preventing unintended proteolysis in plant protein biofactories

Plant Biotechnol J. 2008 Sep;6(7):633-48. doi: 10.1111/j.1467-7652.2008.00344.x. Epub 2008 Apr 28.

Abstract

Numerous reports have been published over the last decade assessing the potential of plants as useful hosts for the heterologous expression of clinically useful proteins. Significant progress has been made, in particular, in optimizing transgene transcription and translation in plants, and in elucidating the complex post-translational modifications of proteins typical of the plant cell machinery. In this article, we address the important issue of recombinant protein degradation in plant expression platforms, which directly impacts on the final yield, homogeneity and overall quality of the resulting protein product. Unlike several more stable and structurally less complex pharmaceuticals, recombinant proteins present a natural tendency to structural heterogeneity, resulting in part from the inherent instability of polypeptide chains expressed in heterologous environments. Proteolytic processing, notably, may dramatically alter the structural integrity and overall accumulation of recombinant proteins in plant expression systems, both in planta during expression and ex planta after extraction. In this article, we describe the current strategies proposed to minimize protein hydrolysis in plant protein factories, including organ-specific transgene expression, organelle-specific protein targeting, the grafting of stabilizing protein domains to labile proteins, protein secretion in natural fluids and the co-expression of companion protease inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Gene Expression
  • Hydrolysis
  • Peptide Hydrolases / genetics
  • Peptide Hydrolases / physiology
  • Plant Proteins / genetics
  • Plant Proteins / physiology
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / metabolism*
  • Protease Inhibitors / chemistry
  • Protein Engineering / methods*
  • Protein Transport
  • Recombinant Proteins / metabolism*
  • Transgenes

Substances

  • Plant Proteins
  • Protease Inhibitors
  • Recombinant Proteins
  • Peptide Hydrolases