Production of a recombinant industrial protein using barley cell cultures

Ritala, Anneli; Wahlström, Eva; Holkeri, Heidi; Hafrén, Anders; Mäkeläinen, Katri; Báez, Julio; Mäkinen, Kristiina; Nuutila, Anna-Maria

doi:10.1016/j.pep.2008.02.013

Cited by 52 publications

(50 citation statements)

References 52 publications

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“…The foldon of a barley-derived CIa1 was also retained. 13 As expected the foldon was removed by pepsin treatment (Figure 4, Lane 2). This observation is consistent with the production of rCIa1 in Pichia, where neither the C-propeptide nor the foldon were removed and subsequent pepsin treatment was required to convert the procollagens to mature collagens.…”

Section: Assembly Of Triple-helical Rcia1 Structuresupporting

confidence: 75%

“…1,7 Previous hosts for recombinant collagen-related proteins (yeast, insect cell culture, and barley cell culture) produced collagen without prolyl hydroxylation indicating a lack of endogenous P4H activity. 1,8,[10][11][12][13] Co-expression of mammalian recombinant P4H with collagen in recombinant yeast enabled prolyl hydroxylation. 7,14,15 Expression of rCIa1 in transgenic tobacco resulted in low hydroxyproline content ($0.5%) 9 that was significantly increased by the co-expression of a mammalian P4H both for CIa1 as well as type III collagen.…”

Section: Introductionmentioning

confidence: 99%

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Purification and characterization of a transgenic corn grain‐derived recombinant collagen type I alpha 1

Zhang

Báez

Pappu³

et al. 2009

Biotechnology Progress

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Corn offers advantages as a transgenic host for producing recombinant proteins required at large volumes (1,000's of tons per year) and low cost (less than US$50/kg) by generating them as co-products of biorefining. We describe the purification and characterization of a corn grain-derived mammalian structural protein having such market characteristics: a full length recombinant collagen type I alpha 1 (rCI alpha 1) chain. Material properties of interest are gelation behavior, which would depend on as yet unverified ability of corn to carry out post-translational prolyl hydroxylation and formation of triple helical conformation. The starting material was grain where the expression of rCI alpha 1 had been directed by an embryo-specific promoter. Purification consisted of extraction at low pH followed by membrane and chromatographic steps to isolate rCI alpha 1 for characterization. The amino acid composition and immunoreactivity of CI alpha 1 was similar to that of an analogous native human CI alpha 1 and to rCI alpha 1 produced by the yeast Pichia pastoris. Tandem mass spectrometry confirmed the primary sequence of the corn-derived rCI alpha 1 with 46% coverage. Fragments of the rCI alpha 1 chains were also observed, possibly caused by endogenous plant proteases. The corn-derived rCI alpha 1 had a low level of prolyl hydroxylation (approximately 1% versus 11%) relative to animal-derived CI alpha 1 and folded into its characteristic triple-helical structure as indicated by its resistance to pepsin digestion below its melting temperature of 26(o)C. The 29 amino acid foldon fused to the C-terminus to initiate triple helix formation was not cleaved from the rCI alpha 1 chains, but could be removed by pepsin treatment.

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Section: Assembly Of Triple-helical Rcia1 Structuresupporting

confidence: 75%

Section: Introductionmentioning

confidence: 99%

Purification and characterization of a transgenic corn grain‐derived recombinant collagen type I alpha 1

Zhang

Báez

Pappu³

et al. 2009

Biotechnology Progress

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“…This is hindered by the requirement for proline hydroxylation, which has proven difficult in E. coli, though expression can be achieved by exploiting folding catalyst over-expression [112] while collagen mimetics of various formats have also been efficiently expressed in E. coli. [113,114] Native collagen is frequently expressed, however, in non-procaryotic expression systems such as yeast, [115] insect [115c] or plant [116] species for purification and utilization.…”

Section: Expression Systems For Recombinant Productsmentioning

confidence: 99%

Adding Functions to Biomaterial Surfaces through Protein Incorporation

et al. 2016

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The concept of biomaterials has evolved from one of inert mechanical supports with a long-term, biologically inactive role in the body into complex matrices that exhibit selective cell binding, promote proliferation and matrix production, and may ultimately become replaced by newly generated tissues in vivo. Functionalization of material surfaces with biomolecules is critical to their ability to evade immunorecognition, interact productively with surrounding tissues and extracellular matrix, and avoid bacterial colonization. Antibody molecules and their derived fragments are commonly immobilized on materials to mediate coating with specific cell types in fields such as stent endothelialization and drug delivery. The incorporation of growth factors into biomaterials has found application in promoting and accelerating bone formation in osteogenerative and related applications. Peptides and extracellular matrix proteins can impart biomolecule- and cell-specificities to materials while antimicrobial peptides have found roles in preventing biofilm formation on devices and implants. In this progress report, we detail developments in the use of diverse proteins and peptides to modify the surfaces of hard biomaterials in vivo and in vitro. Chemical approaches to immobilizing active biomolecules are presented, as well as platform technologies for isolation or generation of natural or synthetic molecules suitable for biomaterial functionalization.

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“…A recent product from this company was human FLT3-ligand, with the gene under the control of the barley HORDEIN D promoter (Erlendsson et al, 2010). Ritala et al (2008) were able to express a codon-optimized version of COLLAGEN I in barley endosperm-derived suspension cells, and showed that the recombinant protein was equivalent to a version produced in Pichia pastoris yeast. The gene was driven by the maize UBIQUITIN-1 promoter and the resulting protein yield was rather low (2-9 µg/l).…”

Section: Human Proteins and Growth Factorsmentioning

confidence: 99%

Genetic Transformation of Triticeae Cereals for Molecular Farming

Hensel¹

2011

Genetic Transformation

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Production of a recombinant industrial protein using barley cell cultures

Cited by 52 publications

References 52 publications

Purification and characterization of a transgenic corn grain‐derived recombinant collagen type I alpha 1

Purification and characterization of a transgenic corn grain‐derived recombinant collagen type I alpha 1

Adding Functions to Biomaterial Surfaces through Protein Incorporation

Genetic Transformation of Triticeae Cereals for Molecular Farming

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