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Planta Med 2017; 83(10): 862-869
DOI: 10.1055/s-0043-103964
Natural Product Chemistry and Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Recombinant Human Acidic Fibroblast Growth Factor (aFGF) Expressed in Nicotiana benthamiana Potentially Inhibits Skin Photoaging

Jang-Ho Ha   *
1   Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
,
Ha-Neul Kim   *
2   Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeonnam, Korea
,
Ki-Beom Moon
1   Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
,
Jae-Heung Jeon
1   Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
,
Dai-Hyun Jung
3   Medical Nanomaterial Institute, Bio-FD&C, Hwasun, Jeonnam 58141, Korea
,
Su-Jung Kim
3   Medical Nanomaterial Institute, Bio-FD&C, Hwasun, Jeonnam 58141, Korea
,
Hugh S. Mason
4   Biodesign Institute, School of Life Sciences, Arizona State University, Tempe, AZ, USA
,
Seo-Yeon Shin
2   Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeonnam, Korea
,
Hyun-Soon Kim
1   Sustainable Resources Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
,
Kyung-Mok Park
2   Department of Pharmaceutical Engineering, Dongshin University, Naju, Jeonnam, Korea
› Author Affiliations
Further Information

Publication History

received 25 September 2016
revised 26 January 2017

accepted 07 February 2017

Publication Date:
01 March 2017 (online)

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Abstract

Responding to the need for recombinant acidic fibroblast growth factor in the pharmaceutical and cosmetic industries, we established a scalable expression system for recombinant human aFGF using transient and a DNA replicon vector expression in Nicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered following Agrobacterium infiltration of N. benthamiana. The optimal time point at which to harvest recombinant human acidic fibroblast growth factor expressing leaves was found to be 4 days post-infiltration, before necrosis was evident. Commassie-stained SDS-PAGE gels of His-tag column eluates, concentrated using a 10 000 molecular weight cut-off column, showed an intense band at the expected molecular weight for recombinant human acidic fibroblast growth factor. An immunoblot confirmed that this band was recombinant human acidic fibroblast growth factor. Up to 10 µg recombinant human-acidic fibroblast growth factor/g of fresh leaves were achieved by a simple affinity purification protocol using protein extract from the leaves of agroinfiltrated N. benthamiana. The purified recombinant human acidic fibroblast growth factor improved the survival rate of UVB-irradiated HaCaT and CCD-986sk cells approximately 89 and 81 %, respectively. N. benthamiana-derived recombinant human acidic fibroblast growth factor showed similar effects on skin cell proliferation and UVB protection compared to those of Escherichia coli-derived recombinant human acidic fibroblast growth factor. Additionally, N. benthamiana-derived recombinant human acidic fibroblast growth factor increased type 1 procollagen synthesis up to 30 % as well as reduced UVB-induced intracellular reactive oxygen species generation in fibroblast (CCD-986sk) cells.

UVB is a well-known factor that causes various types of skin damage and premature aging. Therefore, the present study demonstrated that N. benthamiana-derived recombinant human acidic fibroblast growth factor effectively protects skin cell from UVB, suggesting its potential use as a cosmetic or therapeutic agent against skin photoaging.

Key words

Nicotiana benthamiana - Solanaceae - recombinant human aFGF - transient expression - DNA replicon vector - skin photoaging - UVB irradiation

* These two authors contributed equally to this work.


Supporting Information

A schematic representation of the expression vector used in this study is available as Supporting Information.

 

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