Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants

Andrew Conley, Jussi Joensuu, Alex Richman, Rima Menassa (Corresponding Author)

Research output: Contribution to journalReview ArticleScientificpeer-review

78 Citations (Scopus)

Abstract

For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant‐made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant‐made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin‐like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide ‘VPGXG’ sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant‐based aqueous two‐phase systems. Zera, a domain of the maize seed storage protein γ‐zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density‐based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification.
Original languageEnglish
Pages (from-to)419-433
JournalPlant Biotechnology Journal
Volume9
Issue number4
DOIs
Publication statusPublished - 2011
MoE publication typeA2 Review article in a scientific journal

Fingerprint

protein bodies
Recombinant Proteins
recombinant proteins
Proteins
proteins
fungal proteins
purification methods
zein
seed storage proteins
Seed Storage Proteins
Zein
biopolymers
Artificial Cells
storage proteins
hydrophobicity
Biopolymers
Fungal Proteins
organelles
polypeptides
Organelle Biogenesis

Keywords

  • molecular farming
  • recombinant protein purification
  • elastin-like polypeptides
  • hydrophobins
  • zein
  • protein bodies

Cite this

@article{5f30b169ce444c99a3bdfc3b4584b389,
title = "Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants",
abstract = "For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant‐made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant‐made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin‐like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide ‘VPGXG’ sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant‐based aqueous two‐phase systems. Zera, a domain of the maize seed storage protein γ‐zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density‐based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification.",
keywords = "molecular farming, recombinant protein purification, elastin-like polypeptides, hydrophobins, zein, protein bodies",
author = "Andrew Conley and Jussi Joensuu and Alex Richman and Rima Menassa",
year = "2011",
doi = "10.1111/j.1467-7652.2011.00596.x",
language = "English",
volume = "9",
pages = "419--433",
journal = "Plant Biotechnology Journal",
issn = "1467-7644",
publisher = "Wiley",
number = "4",

}

Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants. / Conley, Andrew; Joensuu, Jussi; Richman, Alex; Menassa, Rima (Corresponding Author).

In: Plant Biotechnology Journal, Vol. 9, No. 4, 2011, p. 419-433.

Research output: Contribution to journalReview ArticleScientificpeer-review

TY - JOUR

T1 - Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants

AU - Conley, Andrew

AU - Joensuu, Jussi

AU - Richman, Alex

AU - Menassa, Rima

PY - 2011

Y1 - 2011

N2 - For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant‐made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant‐made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin‐like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide ‘VPGXG’ sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant‐based aqueous two‐phase systems. Zera, a domain of the maize seed storage protein γ‐zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density‐based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification.

AB - For the past two decades, therapeutic and industrially important proteins have been expressed in plants with varying levels of success. The two major challenges hindering the economical production of plant‐made recombinant proteins include inadequate accumulation levels and the lack of efficient purification methods. To address these limitations, several fusion protein strategies have been recently developed to significantly enhance the production yield of plant‐made recombinant proteins, while simultaneously assisting in their subsequent purification. Elastin‐like polypeptides are thermally responsive biopolymers composed of a repeating pentapeptide ‘VPGXG’ sequence that are valuable for the purification of recombinant proteins. Hydrophobins are small fungal proteins capable of altering the hydrophobicity of their respective fusion partner, thus enabling efficient purification by surfactant‐based aqueous two‐phase systems. Zera, a domain of the maize seed storage protein γ‐zein, can induce the formation of protein storage bodies, thus facilitating the recovery of fused proteins using density‐based separation methods. These three novel protein fusion systems have also been shown to enhance the accumulation of a range of different recombinant proteins, while concurrently inducing the formation of protein bodies. The packing of these fusion proteins into protein bodies may exclude the recombinant protein from normal physiological turnover. Furthermore, these systems allow for quick, simple and inexpensive nonchromatographic purification of the recombinant protein, which can be scaled up to industrial levels of protein production. This review will focus on the similarities and differences of these artificial storage organelles, their biogenesis and their implication for the production of recombinant proteins in plants and their subsequent purification.

KW - molecular farming

KW - recombinant protein purification

KW - elastin-like polypeptides

KW - hydrophobins

KW - zein

KW - protein bodies

U2 - 10.1111/j.1467-7652.2011.00596.x

DO - 10.1111/j.1467-7652.2011.00596.x

M3 - Review Article

VL - 9

SP - 419

EP - 433

JO - Plant Biotechnology Journal

JF - Plant Biotechnology Journal

SN - 1467-7644

IS - 4

ER -