Protein Expression Systems: Why Soybean Seeds?

Bost, Kenneth L.; Piller, Kenneth J.

doi:10.5772/15376

Cited by 8 publications

(10 citation statements)

References 35 publications

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“…Although RNA-seq is focused on the functional segments of the genome and does not always capture regulatory regions such as promoters or nontranscribed regions (e.g., methylated bases), one aspect of the resulting effects of polymorphisms in these regions can be directly witnessed through examination of gene expression levels. Because the soybean expression system described previously by our lab [5, 7, 8, 13, 14, 39] specifically targets seed tissue for recombinant protein accumulation, it was of great interest to identify possible sequence alterations that may have resulted in gene expression or protein structure variations in seeds, which to date has not been investigated.…”

Section: Discussionmentioning

confidence: 99%

“…Over the past 10 years, our laboratory has focused on Glycine max as a model system for the generation and storage of recombinant proteins that are currently expensive to manufacture and difficult to procure or synthesize in bacterial and cell-based cultures and as oral vaccine candidates by targeting gut-associated lymphoid tissues (GALT) for immune stimulation or suppression [5, 8, 10–13]. Advantages for using soybean as an expression system include high protein content and yield, self-fertilization to simplify the generation of homozygous transformants, and the stability of endogenous proteins in seed tissues, which we have reviewed in depth previously [14]. …”

Section: Introductionmentioning

confidence: 99%

“…Soluble protein extraction from soy seed is also straightforward and has proven to be efficient for the generation and purification of vaccine antigens, therapeutic proteins, and diagnostic peptides [7–9]. This makes soybean an ideal candidate platform as a bioreactor for the generation, accumulation, and long-term storage of plant-based biologics [14], removing the trade-off dichotomy of either high yield (leaf tissue expression) or long-term stability (seed tissue).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Transcript Polymorphism Rates in Soybean Seed Tissue Are Increased in a Single Transformant ofGlycine max

Lambirth

Whaley

Schlueter

et al. 2016

International Journal of Plant Genomics

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Transgenic crops have been utilized for decades to enhance agriculture and more recently have been applied as bioreactors for manufacturing pharmaceuticals. Recently, we investigated the gene expression profiles of several in-house transgenic soybean events, finding one transformant group to be consistently different from our controls. In the present study, we examined polymorphisms and sequence variations in the exomes of the same transgenic soybean events. We found that the previously dissimilar soybean line also exhibited markedly increased levels of polymorphisms within mRNA transcripts from seed tissue, many of which are classified as gene expression modifiers. The results from this work will direct future investigations to examine novel SNPs controlling traits of great interest for breeding and improving transgenic soybean crops. Further, this study marks the first work to investigate SNP rates in transgenic soybean seed tissues and demonstrates that while transgenesis may induce abundant unanticipated changes in gene expression and nucleotide variation, phenotypes and overall health of the plants examined remained unaltered.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Transcript Polymorphism Rates in Soybean Seed Tissue Are Increased in a Single Transformant ofGlycine max

Lambirth

Whaley

Schlueter

et al. 2016

International Journal of Plant Genomics

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“…Cellobiohydrolase II (Cel6A) targeted to maize endosperm, represents 30% of seed TSP. Recombinant protein yield in soybean can reach 4% of TSP [80]. Since natural NK is produced from fermented soybeans, it represents an ideal platform for recombinant NK production.…”

Section: Plants As Potential Factories For Nattokinase Productionmentioning

confidence: 99%

Nattokinase: An Oral Antithrombotic Agent for the Prevention of Cardiovascular Disease

Weng

Yao

Sparks

et al. 2017

IJMS

144

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Natto, a fermented soybean product, has been consumed as a traditional food in Japan for thousands of years. Nattokinase (NK), a potent blood-clot dissolving protein used for the treatment of cardiovascular diseases, is produced by the bacterium Bacillus subtilis during the fermentation of soybeans to produce Natto. NK has been extensively studied in Japan, Korea, and China. Recently, the fibrinolytic (anti-clotting) capacity of NK has been recognized by Western medicine. The National Science Foundation in the United States has investigated and evaluated the safety of NK. NK is currently undergoing a clinical trial study (Phase II) in the USA for atherothrombotic prevention. Multiple NK genes have been cloned, characterized, and produced in various expression system studies. Recombinant technology represents a promising approach for the production of NK with high purity for its use in antithrombotic applications. This review covers the history, benefit, safety, and production of NK. Opportunities for utilizing plant systems for the large-scale production of NK, or for the production of edible plants that can be used to provide oral delivery of NK without extraction and purification are also discussed.

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“…Transgenic Soybean Production of Bioactive Human Epidermal Growth Factor (EGF) is to develop a biopharma platform that is broadly acceptable for food and feed delivery but can be lightly processed to preserve bioactivity and can be massively scaled to maximize the distribution potential of the product. Soybean is a potentially useful biopharma platform that could have broad application in both food and feed end uses [29,30,56,57]. Soybean has been demonstrated as a platform to produce heterologous proteins at a standard that far exceeds the levels typically needed for biopharma [31].…”

Section: Soybean Seed Platform For Biopharmamentioning

confidence: 99%

Transgenic Soybean Production of Bioactive Human Epidermal Growth Factor (EGF)

Schmidt

Erwin

et al. 2016

PLoS ONE

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Necrotizing enterocolitis (NEC) is a devastating condition of premature infants that results from the gut microbiome invading immature intestinal tissues. This results in a life-threatening disease that is frequently treated with the surgical removal of diseased and dead tissues. Epidermal growth factor (EGF), typically found in bodily fluids, such as amniotic fluid, salvia and mother’s breast milk, is an intestinotrophic growth factor and may reduce the onset of NEC in premature infants. We have produced human EGF in soybean seeds to levels biologically relevant and demonstrated its comparable activity to commercially available EGF. Transgenic soybean seeds expressing a seed-specific codon optimized gene encoding of the human EGF protein with an added ER signal tag at the N’ terminal were produced. Seven independent lines were grown to homozygous and found to accumulate a range of 6.7 +/- 3.1 to 129.0 +/- 36.7 μg EGF/g of dry soybean seed. Proteomic and immunoblot analysis indicates that the inserted EGF is the same as the human EGF protein. Phosphorylation and immunohistochemical assays on the EGF receptor in HeLa cells indicate the EGF protein produced in soybean seed is bioactive and comparable to commercially available human EGF. This work demonstrates the feasibility of using soybean seeds as a biofactory to produce therapeutic agents in a soymilk delivery platform.

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Protein Expression Systems: Why Soybean Seeds?

Cited by 8 publications

References 35 publications

Transcript Polymorphism Rates in Soybean Seed Tissue Are Increased in a Single Transformant ofGlycine max

Transcript Polymorphism Rates in Soybean Seed Tissue Are Increased in a Single Transformant ofGlycine max

Nattokinase: An Oral Antithrombotic Agent for the Prevention of Cardiovascular Disease

Transgenic Soybean Production of Bioactive Human Epidermal Growth Factor (EGF)

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