Seed-Specific Expression of Spider Silk Protein Multimers Causes Long-Term Stability

Weichert, Nicola; Hauptmann, Valeska; Helmold, Christine; Conrad, Udo

doi:10.3389/fpls.2016.00006

Cited by 33 publications

(25 citation statements)

References 58 publications

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“…Synthetic FLAG spidroin sequence, flanked by intein splicing elements and directed to the ER in tobacco seeds, produced multimers of up to 450 kDa with an estimated yield of 20-190 mg per kg seed (fresh weight). The recombinant FLAG proteins produced in tobacco seeds are stable at 15°C for one year (Weichert et al, 2016). Another example of in vitro PTM is multimerization of rMaSp1-100x ELP fusion protein tagged with lysine or glutamine by transglutaminase, which can produce near native sized spidroin-like recombinant protein (Weichert et al, 2014).…”

Section: Post-translational Modification (Ptm) and In Vitro Modificationmentioning

confidence: 99%

“…Recombinant spidroin-like proteins are stable under normal seed storage conditions (Weichert et al, 2016). An economic advantage of transgenic crop seeds producing recombinant protein for medicinal usage is that subjects can be fed directly thereby bypassing cumbersome protein purification.…”

Section: Diabetesmentioning

confidence: 99%

“…However, recombinant tropoelastin degraded into two smaller molecular weight peptides when leaf extracts were frozen (Abdelghani et al, 2015). As the instability of protein going through freeze thaw of the leaf tissues will create a problem for potential harvesting and storage, recombinant tropoelastin directed to seeds may be more suitable, since recombinant protein can be stably stored in the seeds (Weichert et al, 2016).…”

Section: Elastin-based Scaffold Protein Produced From Plantsmentioning

confidence: 99%

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Advances in Plant-Derived Scaffold Proteins

2020

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Scaffold proteins form critical biomatrices that support cell adhesion and proliferation for regenerative medicine and drug screening. The increasing demand for such applications urges solutions for cost effective and sustainable supplies of hypoallergenic and biocompatible scaffold proteins. Here, we summarize recent efforts in obtaining plantderived biosynthetic spider silk analogue and the extracellular matrix protein, collagen. Both proteins are composed of a large number of tandem block repeats, which makes production in bacterial hosts challenging. Furthermore, post-translational modification of collagen is essential for its function which requires co-transformation of multiple copies of human prolyl 4-hydroxylase. We discuss our perspectives on how the GAANTRY system could potentially assist the production of native-sized spider dragline silk proteins and prolyl hydroxylated collagen. The potential of recombinant scaffold proteins in drug delivery and drug discovery is also addressed.

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Section: Post-translational Modification (Ptm) and In Vitro Modificationmentioning

confidence: 99%

Section: Diabetesmentioning

confidence: 99%

Section: Elastin-based Scaffold Protein Produced From Plantsmentioning

confidence: 99%

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Advances in Plant-Derived Scaffold Proteins

2020

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“…The presence of endophytes in seed batches and the identity of endophyte strains were confirmed using a Competitive Allele Specific PCR (KASP) bioassay [39]. KASP primers were previously designed and generated by AgriBio and GeneWorks based on allelic variation of single nucleotide polymorphisms (SNP) that differentiate endophytes from one another (SE, AR1, AR37, NEA2 and NEA6).…”

Section: Endophytesmentioning

confidence: 99%

Novel bioassay to assess antibiotic effects of fungal endophytes on aphids

et al. 2020

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Perennial ryegrass is an important feed base for the dairy and livestock industries around the world. It is often infected with mutualistic fungal endophytes that confer protection to the plant against biotic and abiotic stresses. Bioassays that test their antibiotic effect on invertebrates are varied and range from excised leaves to whole plants. The aim of this study was to design and validate a "high-throughput" in-planta bioassay using 7-day-old seedlings confined in small cups, allowing for rapid assessments of aphid life history to be made while maintaining high replication and treatment numbers. Antibiosis was evaluated on the foliar and the root aphid species; Diuraphis noxia (Mordvilko) and Aploneura lentisci (Passerini) feeding on a range of perennial ryegrass-Epichloë festucae var. Lolii endophyte symbiota. As expected, both D. noxia and A. lentisci reared on endophyte-infected plants showed negatively affected life history traits by comparison to non-infected plants. Both species exhibited the highest mortality at the nymphal stage with an average total mortality across all endophyte treatments of 91% and 89% for D. noxia and A. lentisci respectively. Fecundity decreased significantly on all endophyte treatments with an average total reduction of 18% and 16% for D. noxia and A. lentisci respectively by comparison to non-infected plants.Overall, the bioassay proved to be a rapid method of evaluating the insecticidal activity of perennial ryegrass-endophyte symbiota on aphids (nymph mortality could be assessed in as little as 24 and 48 hours for D. noxia and A. lentisci respectively). This rapid and simple approach can be used to benchmark novel grass-endophyte symbiota on a range of aphid species that feed on leaves of plants, however we would caution that it may not be suitable for the assessment of root-feeding aphids, as this species exhibited relatively high mortality on the control as well.

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“…Stable expression yields up to 10% of total seed proteins have been achieved [170,175]. Products include therapeutic proteins such as antibodies [176], vaccines [177], and cytokines [178]; industrial enzymes such as trypsin [179], phytase [180], and cellulase [181]; and biopolymers such as spider silk protein [182]. See recent review [173].…”

Section: Stable Expression: Seed Basedmentioning

confidence: 99%

Platforms for Plant-Based Protein Production

Xu¹,

Towler²,

Weathers³

2016

Bioprocessing of Plant in Vitro Systems

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Plant molecular farming depends on a diversity of plant systems for production of useful recombinant proteins. These proteins include protein biopolymers, industrial proteins and enzymes, and therapeutic proteins. Plant production systems include microalgae, cells, hairy roots, moss, and whole plants with both stable and transient expression. Production processes involve a narrowing diversity of bioreactors for cell, hairy root, microalgae, and moss cultivation. For whole plants, both field and automated greenhouse cultivation methods are used with

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Seed-Specific Expression of Spider Silk Protein Multimers Causes Long-Term Stability

Cited by 33 publications

References 58 publications

Advances in Plant-Derived Scaffold Proteins

Advances in Plant-Derived Scaffold Proteins

Novel bioassay to assess antibiotic effects of fungal endophytes on aphids

Platforms for Plant-Based Protein Production

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