TY - JOUR

T1 - Modulation of Barley (Hordeum vulgare L.) Grain Protein Sink-Source Relations Towards Human Epidermal Growth Factor Instead of B-hordein Storage Protein

AU - Panting, Michael

AU - Holme, Inger

AU - Björnsson, Jón Már

AU - Brinch-Pedersen, Henrik

PY - 2020/10/13

Y1 - 2020/10/13

N2 - Seeds have evolutionarily developed to store protein without immediately degrading it and constitute ideal tissues for recombinant protein storage. Unfortunately, the production of recombinant protein in seeds is compromised by low yield as compared to other heterologous expression systems. In order to improve the yield of the human epidermal growth factor (EGF) in barley, protein sink-source relations in the developing grain were modulated towards EGF instead of the barley storage protein. The EGF gene, under the control of a B-hordein and a seed-specific oat globulin promoter, was introduced by crossing EGF lines into the Risø 56 mutant deficient in B-hordein storage protein synthesis. Offspring plants were analysed for EGF and Hordein expression and for expression of the unfolded protein response (UPR) genes PDI and CRT to monitor changes in ER stress levels. EGF content was increased significantly in the mature grain of homozygous offspring and PDI and CRT gene expressions were upregulated. We demonstrate, for the first time in barley, that replacement of an abundant seed storage protein with a specific heterologous protein driven by the promoter of the removed gene can accelerate the production of a specific heterologous protein in barley grains.

AB - Seeds have evolutionarily developed to store protein without immediately degrading it and constitute ideal tissues for recombinant protein storage. Unfortunately, the production of recombinant protein in seeds is compromised by low yield as compared to other heterologous expression systems. In order to improve the yield of the human epidermal growth factor (EGF) in barley, protein sink-source relations in the developing grain were modulated towards EGF instead of the barley storage protein. The EGF gene, under the control of a B-hordein and a seed-specific oat globulin promoter, was introduced by crossing EGF lines into the Risø 56 mutant deficient in B-hordein storage protein synthesis. Offspring plants were analysed for EGF and Hordein expression and for expression of the unfolded protein response (UPR) genes PDI and CRT to monitor changes in ER stress levels. EGF content was increased significantly in the mature grain of homozygous offspring and PDI and CRT gene expressions were upregulated. We demonstrate, for the first time in barley, that replacement of an abundant seed storage protein with a specific heterologous protein driven by the promoter of the removed gene can accelerate the production of a specific heterologous protein in barley grains.

KW - Barley heterologous expression

KW - Molecular pharming

KW - B-hordein

KW - Human EGF

KW - Unfolded protein response (UPR)

U2 - 10.1007/s12033-020-00279-3

DO - 10.1007/s12033-020-00279-3

M3 - Journal article

C2 - 33051823

JO - Molecular Biotechnology

JF - Molecular Biotechnology

SN - 1073-6085

ER -