High-level transient production of a protease-resistant mutant form of human basic fibroblast growth factor in Nicotiana benthamiana leaves
Issued Date
2022-01-01
Resource Type
ISSN
13424580
eISSN
13476114
Scopus ID
2-s2.0-85138731047
Journal Title
Plant Biotechnology
Volume
39
Issue
3
Start Page
291
End Page
301
Rights Holder(s)
SCOPUS
Bibliographic Citation
Plant Biotechnology Vol.39 No.3 (2022) , 291-301
Suggested Citation
Macauyag E.A., Kajiura H., Ohashi T., Misaki R., Fujiyama K. High-level transient production of a protease-resistant mutant form of human basic fibroblast growth factor in Nicotiana benthamiana leaves. Plant Biotechnology Vol.39 No.3 (2022) , 291-301. 301. doi:10.5511/plantbiotechnology.22.0628a Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/87122
Title
High-level transient production of a protease-resistant mutant form of human basic fibroblast growth factor in Nicotiana benthamiana leaves
Author(s)
Author's Affiliation
Other Contributor(s)
Abstract
The human basic fibroblast growth factor (bFGF) is a protein that plays a pivotal role in cellular processes like cell proliferation and development. As a result, it has become an important component in cell culture systems, with applications in biomedical engineering, cosmetics, and research. Alternative production techniques, such as transient production in plants, are becoming a feasible option as the demand continues to grow. High-level bFGF production was achieved in this study employing an optimized Agrobacterium-mediated transient expression system, which yielded about a 3-fold increase in production over a conventional system. This yield was further doubled at about 185 µg g−1 FW using a mutant protease-resistant version that degraded/aggregated at a three-fold slower rate in leaf crude extracts. To achieve a pure product, a two-step purification technique was applied. The capacity of the pure protease-resistant bFGF (PRbFGF) to stimulate cell proliferation was tested and was found to be comparable to that of E. coli-produced bFGF in HepG2 and CHO-K1 cells. Overall, this study demonstrates a high-level transient production system of functional PRbFGF in N. benthamiana leaves as well as an efficient tag-less purification technique of leaf crude extracts.