Production of bioactive, post-translationally modified, heterotrimeric, human recombinant type-I collagen in transgenic tobacco.
Collagen's biocompatibility, biodegradability and low immunogenicity render it advantageous for extensive application in pharmaceutical or biotechnological disciplines. However, typical collagen extraction from animal or cadaver sources harbors risks including allergenicity and potential sample contamination with pathogens. In this work, two human genes encoding recombinant heterotrimeric collagen type I (rhCOL1) were successfully coexpressed in tobacco plants with the human prolyl-4-hydroxylase (P4H) and lysyl hydroxylase 3 (LH3) enzymes, responsible for key posttranslational modifications of collagen. Plants coexpressing all five vacuole-targeted proteins generated intact procollagen yields of approximately 2% of the extracted total soluble proteins. Plant-extracted rhCOL1 formed thermally stable triple helical structures and demonstrated biofunctionality similar to human tissue-derived collagen supporting binding and proliferation of adult peripheral blood-derived endothelial progenitor-like cells. Through a simple, safe and scalable method of rhCOL1 production and purification from tobacco plants, this work broadens the potential applications of human recombinant collagen in regenerative medicine.
Stein
Hanan
H
Collplant Ltd., 3 Sapir St, Weizmann Science Park, PO Box 4132, Ness-Ziona 74140, Israel.
Wilensky
Michal
M
Tsafrir
Yehuda
Y
Rosenthal
Michal
M
Amir
Rachel
R
Avraham
Tal
T
Ofir
Keren
K
Dgany
Or
O
Yayon
Avner
A
Shoseyov
Oded
O
D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D030821:Plants, Genetically Modified; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D011393:Procollagen-Proline Dioxygenase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins; D014026:Nicotiana / Q000235:genetics
D024042:Collagen Type I; D011994:Recombinant Proteins; C475767:PLOD3 protein, human; D011393:Procollagen-Proline Dioxygenase; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase