Expression of biologically active anti-thrombosis protein lumbrokinase in edible sunflower seed kernel

Guan, Chunfeng; Du, Xilong; Wang, Gang; Ji, Jing; Chen, Jin; Li, Xiaozhou

doi:10.1007/s13562-013-0209-7

Cited by 15 publications

(13 citation statements)

References 20 publications

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“…Using PMF to create a vegetable, seed or fruit health supplement could be a practical alternative to using PMF to develop a processed pharmaceutical drug. Guan et al (2014) [ 57 , 58 ] expressed lumbrokinase, an anti-thrombotic enzyme from earthworm, in sunflower kernels. Mice and rats that were fed the transgenic kernels exhibited a strong degradation of blood clots [ 58 ].…”

Section: Pmf Products For Use As Topical Applications and Health Smentioning

confidence: 99%

“…Guan et al (2014) [ 57 , 58 ] expressed lumbrokinase, an anti-thrombotic enzyme from earthworm, in sunflower kernels. Mice and rats that were fed the transgenic kernels exhibited a strong degradation of blood clots [ 58 ]. Unlike a vaccine or a therapeutic protein, lumbrokinase has been widely sold and used as a health supplement to dissolve blood clots and maintain healthy cardiovascular function in people.…”

Section: Pmf Products For Use As Topical Applications and Health Smentioning

confidence: 99%

“…When these proteins were targeted to in seeds, however, cell necrosis was not observed and the purified proteins exhibited the ability to dissolve fibrin and blood clots (unpublished data). Therefore, targeting the production of PMF-based products to seeds is becoming an attractive alternative [ 8 , 57 , 58 , 90 , 91 , 92 , 93 , 94 ].…”

Section: Pmf Production Platformsmentioning

confidence: 99%

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Plants as Factories for Human Pharmaceuticals: Applications and Challenges

Yao

Weng

Dickey

et al. 2015

IJMS

219

162

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Plant molecular farming (PMF), defined as the practice of using plants to produce human therapeutic proteins, has received worldwide interest. PMF has grown and advanced considerably over the past two decades. A number of therapeutic proteins have been produced in plants, some of which have been through pre-clinical or clinical trials and are close to commercialization. Plants have the potential to mass-produce pharmaceutical products with less cost than traditional methods. Tobacco-derived antibodies have been tested and used to combat the Ebola outbreak in Africa. Genetically engineered immunoadhesin (DPP4-Fc) produced in green plants has been shown to be able to bind to MERS-CoV (Middle East Respiratory Syndrome), preventing the virus from infecting lung cells. Biosafety concerns (such as pollen contamination and immunogenicity of plant-specific glycans) and costly downstream extraction and purification requirements, however, have hampered PMF production from moving from the laboratory to industrial application. In this review, the challenges and opportunities of PMF are discussed. Topics addressed include; transformation and expression systems, plant bioreactors, safety concerns, and various opportunities to produce topical applications and health supplements.

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Section: Pmf Products For Use As Topical Applications and Health Smentioning

confidence: 99%

Section: Pmf Products For Use As Topical Applications and Health Smentioning

confidence: 99%

Section: Pmf Production Platformsmentioning

confidence: 99%

See 1 more Smart Citation

Plants as Factories for Human Pharmaceuticals: Applications and Challenges

Yao

Weng

Dickey

et al. 2015

IJMS

219

162

View full text Add to dashboard Cite

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“…Additionally, plant systems also enable PTMs which can impact protein behavior and increase protein function [ 12 , 13 ]. To our knowledge, there are only three previous reports that have used plant systems to produce LKs [ 14 – 16 ]. Ko et al [ 16 ] introduced a LK gene from the earthworm Eisenia andrei into tobacco plants.…”

Section: Introductionmentioning

confidence: 99%

“…Although high levels of mRNA expression were observed, no enzymatic activity was detected in that study. In 2014, research group successfully expressed a biologically active antithrombotic LK, EFE 3-1 from L. rubellus , in sunflower seed kernels [ 14 , 15 ]. The recombinant LKs yield at 5.1 [ 14 ] and 5.42 g/kg [ 15 ] in recombinant sunflower seeds, respectively.…”

Section: Introductionmentioning

confidence: 99%

Transient Expression of Lumbrokinase (PI239) in Tobacco (Nicotiana tabacum) Using a Geminivirus‐Based Single Replicon System Dissolves Fibrin and Blood Clots

Dickey

Wang

Cooper

et al. 2017

Evidence-Based Complementary and Alternative Medicine

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Lumbrokinases, a group of fibrinolytic enzymes extracted from earthworm, have been widely used to prevent and treat various cardiovascular diseases. They specifically target fibrin to effectively degrade thrombi without major side effects. Plant expression systems are becoming potential alternative expression platforms for producing pharmaceutical proteins. In this work, a lumbrokinase (PI239) was produced from a plant system. Both wild-type (WT) and plant codon-optimized (OP) PI239 gene sequences were synthesized and cloned into a geminivirus-based single-vector DNA replicon system. Both vectors were independently expressed in tobacco (Nicotiana tabacum) leaves transiently by agroinfiltration. Overexpressed PI239 resulted in sudden tissue necrosis 3 days after infiltration. Remaining proteins were purified through His-tag affinity chromatography and analyzed with SDS-PAGE and Western blot methods. Purified PI239 successfully degraded artificial fibrin with relative activity of 13,400 U/mg when compared with commercial lumbrokinase product. In vitro tests demonstrated that plant-derived PI239 dissolved human blood clots and that the plant expression system is capable of producing functional PI239.

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