Transgenic Soybean Production of Bioactive Human Epidermal Growth Factor (EGF).
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.
School of Plant Sciences, University of Arizona, Tucson, Arizona, United States of America.
Yonghua
Y
He
School of Plant Sciences, University of Arizona, Tucson, Arizona, United States of America.
Monica A
MA
Schmidt
St. Louis Children's Hospital and Washington University School of Medicine, St. Louis, Missouri, United States of America.
Christopher
C
Erwin
St. Louis Children's Hospital and Washington University School of Medicine, St. Louis, Missouri, United States of America.
Jun
J
Guo
St. Louis Children's Hospital and Washington University School of Medicine, St. Louis, Missouri, United States of America.
Raphael
R
Sun
School of Animal & Comparative Biomedical Sciences, University of Arizona, Tucson, Arizona, United States of America.
Ken
K
Pendarvis
St. Louis Children's Hospital and Washington University School of Medicine, St. Louis, Missouri, United States of America.
Brad W
BW
Warner
School of Plant Sciences, University of Arizona, Tucson, Arizona, United States of America.
Eliot M
EM
Herman
D020345:Enterocolitis, Necrotizing / Q000235:genetics* / Q000473:pathology; D004815:Epidermal Growth Factor / Q000008:administration & dosage / Q000096:biosynthesis*; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism; D000069196:Gastrointestinal Microbiome / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D010766:Phosphorylation; D030821:Plants, Genetically Modified / Q000235:genetics*; D012639:Seeds / Q000235:genetics; D013025:Glycine max / Q000235:genetics*
D004815:Epidermal Growth Factor; D066246:ErbB Receptors