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Plant Expression Systems for Bioproduct Production

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A special issue of Plants (ISSN 2223-7747).

Deadline for manuscript submissions: closed (31 December 2019) | Viewed by 33635

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Special Issue Editor

Dr. Hyun-Soon Kim

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Guest Editor

Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Korea
Interests: plant biotechnology; plant gene expression; plant molecular farming; plant-derived vaccine; virus-like particle; plant cell culture; plant regeneration
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Special Issue Information

Dear Colleagues,

Plant-based expression systems for bioproducts, including recombinant pharmaceutical proteins, have emerged as an acceptable alternative to conventional expression platforms such as bacterial, yeast, and animal cell. The benefits or advantages cited most often are rapid scalability with transgenic plants or transient expression, higher stability of recombinant proteins, safety due to lack of harmful substances, and capability of producing proteins with desired post-translational modifications.

Due to several significant events, such as ElelysoTM (Protalix, Carmiel, Israel), non-Hodgkin’s lymphoma vaccines (Large Scale Biology, Icon Genetics/Bayer, Halle, Germany), the influenza serotype H1N1 and H5N1 vaccines (Fraunhofer CMB, Newark, DE, and Medicago, Quebec, QC, Canada), and ZMappTM cocktail (Mapp Biotherapeutics, San Diego, CA), readers around the world have become very interested in this plant molecular farming area.

Plant expression systems still have several limitations to be overcome to improve protein yields/quality. The level of protein expression is a critical factor in plant molecular farming, and this level varies according to the plant species and the organs involved and target protein of interest. The expression/production of recombinant native and engineered proteins is a complicated procedure that requires an inter- and multi-disciplinary effort including immunology, genomics, bioinformatics, protein structure, etc. Possible topics in this issue cover research in important plant resources, affecting factors, and the recombinant-protein expression techniques relevant to the plant molecular farming process. Further, this Special Issue will highlight recent advances that have been achieved in plant-based bioproduct production platforms in terms of expression strategies, product yields, and downstream processing development.

I wish to thank all authors for their contributions to this Special issue.

Dr. Hyun-Soon Kim
Guest Editor

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Keywords

plant-based expression system
molecular farming/pharming
recombinant protein
biopharmaceuticals
bioproduct
gene expression
transgenic plant
transient expression

Published Papers (5 papers)

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Research

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14 pages, 2979 KiB

Open AccessArticle

Agrobacterium-Mediated Transformation of Chrysanthemum with Artemisinin Biosynthesis Pathway Genes

by Aleksey Firsov, Tatiana Mitiouchkina, Lyubov Shaloiko, Alexander Pushin, Alexander Vainstein and Sergey Dolgov

Plants 2020, 9(4), 537; https://doi.org/10.3390/plants9040537 - 21 Apr 2020

Cited by 13 | Viewed by 4873

Abstract

Artemisinin-based drugs are the most effective medicine for the malaria treatment. To date, the main method of artemisinin production is its extraction from wormwood plants Artemisia annua L. Due to the limitation of this source, considerable efforts are now directed to the development of methods for artemisinin production using heterologous expression systems. Artemisinin is a sesquiterpene lactone, synthesized through the cyclization of farnesyl diphosphate involved in other sesquiterpene biosynthetic systems. Chrysanthemum species as well as A. annua, belong to Asteraceae family, and had been characterized by containing highly content of sesquiterpenes and their precursors. This makes chrysanthemum a promising target for the production of artemisinin in heterologous host plants. Chrysanthemum (C. morifolium Ramat.) was transformed by Agrobacterium tumefaciens carrying with the binary vectors p1240 and p1250, bearing artemisinin biosynthesis genes coding: amorpha-4,11-diene synthase, artemisinic aldehyde Δ11(13) reductase, amorpha-4,11-diene monooxygenase (p1240 was targeted to the mitochondria and p1250 was targeted to the cytosol), cytochrome P450 reductase from A. annua, as well as yeast truncated 3-hydroxy-3-methylglutarylcoenzyme A reductase. This study obtained 8 kanamycin-resistant lines after transformation with the p1240 and 2 lines from p1250. All target genes were detected in 2 and 1 transgenic lines of the 2 vectors. The transformation frequency of all target genes were 0.33% and 0.17% for p1240 and p1250, relative to the total transformed explant numbers. RT-PCR analysis revealed the transcription of all transferred genes in two lines obtained after transformation with the p1240 vector, confirming the possibility of transferring genetic modules encoding entire biochemical pathways into the chrysanthemum genome. This holds promise for the development of a chrysanthemum-based expression system to produce non-protein substances, such as artemisinin. Full article

(This article belongs to the Special Issue Plant Expression Systems for Bioproduct Production)

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9 pages, 1382 KiB

Open AccessArticle

Fast Track to Discover Novel Promoters in Rice

by Yo-Han Yoo, Yu-Jin Kim, Sunok Moon, Yun-Shil Gho, Woo-Jong Hong, Eui-Jung Kim, Xu Jiang and Ki-Hong Jung

Plants 2020, 9(1), 125; https://doi.org/10.3390/plants9010125 - 18 Jan 2020

Cited by 1 | Viewed by 3386

Abstract

Promoters are key components for the application of biotechnological techniques in crop plants. Reporter genes such as GUS or GFP have been used to test the activity of promoters for diverse applications. A huge number of T-DNAs carrying promoterless GUS near their right borders have been inserted into the rice genome, and 105,739 flanking sequence tags from rice lines with this T-DNA insertion have been identified, establishing potential promoter trap lines for 20,899 out of 55,986 genes in the rice genome. Anatomical meta-expression data and information on abiotic stress related to these promoter trap lines enable us to quickly identify new promoters associated with various expression patterns. In the present report, we introduce a strategy to identify new promoters in a very short period of time using a combination of meta-expression analysis and promoter trap lines. Full article

(This article belongs to the Special Issue Plant Expression Systems for Bioproduct Production)

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12 pages, 1925 KiB

Open AccessArticle

Recombinant Human Dentin Matrix Protein 1 (hDMP1) Expressed in Nicotiana benthamiana Potentially Induces Osteogenic Differentiation

by Aktsar Roskiana Ahmad, Pornjira Kaewpungsup, Narach Khorattanakulchai, Kaewta Rattanapisit, Prasit Pavasant and Waranyoo Phoolcharoen

Plants 2019, 8(12), 566; https://doi.org/10.3390/plants8120566 - 3 Dec 2019

Cited by 9 | Viewed by 3357

Abstract

Inductive molecules are critical components for successful bone tissue engineering. Dentin matrix protein-1 (DMP1), a non-collagenous protein in the bone matrix, has been shown to play roles in osteogenic differentiation and phosphate homeostasis. This study aimed to produce recombinant human dentin matrix protein-1 (hDMP1) in Nicotiana benthamiana and investigated the ability of this plant-produced DMP1 to induce osteogenesis in human periodontal ligament stem cells (hPDLSCs). The hDMP1 gene was cloned into the geminiviral vector for transient expression in N. benthamiana. We found that hDMP1 was transiently expressed in N. benthamiana leaves and could be purified by ammonium sulphate precipitation followed by nickel affinity chromatography. The effects of hDMP1 on the induction of cell proliferation and osteogenic differentiation were investigated. The results indicated that plant-produced hDMP1 could induce the cell proliferation of hPDLSCs and increase the expression levels of osteogenic genes, including osterix (OSX), type I collagen (COL1), bone morphogenetic protein-2 (BMP2), and Wnt3a. Moreover, the plant-produced hDMP1 promoted calcium deposition in hPDLSCs as determined by alizarin red S staining. In conclusion, our results indicated that plant-produced hDMP1 could induce osteogenic differentiation in hPDLSCs and could potentially be used as a bone inducer in bone tissue engineering. Full article

(This article belongs to the Special Issue Plant Expression Systems for Bioproduct Production)

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10 pages, 2397 KiB

Open AccessArticle

Plant-Produced Anti-Enterovirus 71 (EV71) Monoclonal Antibody Efficiently Protects Mice Against EV71 Infection

by Kaewta Rattanapisit, Zhang Chao, Konlavat Siriwattananon, Zhong Huang and Waranyoo Phoolcharoen

Plants 2019, 8(12), 560; https://doi.org/10.3390/plants8120560 - 1 Dec 2019

Cited by 26 | Viewed by 4880

Abstract

Enterovirus 71 (EV71) is the main causative agent of severe hand-foot-mouth disease. EV71 affects countries mainly in the Asia-Pacific region, which makes it unattractive for pharmaceutical companies to develop drugs or vaccine to combat EV71 infection. However, development of these drugs and vaccines is vital to protect younger generations. This study aims to develop a specific monoclonal antibody (mAb) to EV71 using a plant platform, which is a cost-effective and scalable production technology. A previous report showed that D5, a murine anti-EV71 mAb, binds to VP1 protein of EV71, potently neutralizes EV71 in vitro, and effectively protects mice against EV71 infection. Herein, plant-produced chimeric D5 (cD5) mAb, variable regions of murine D5 antibody linked with constant regions of human IgG1, was transiently expressed in Nicotiana benthamiana using geminiviral vectors. The antibody was expressed at high levels within six days of infiltration. Plant-produced cD5 retained its in vitro high-affinity binding and neutralizing activity against EV71. Furthermore, a single dose (10 µg/g body weight) of plant-produced cD5 mAb offered 100% protection against infection in mice after a lethal EV71 challenge. Therefore, our results showed that plant-produced anti-EV71 mAb is an effective, safe, and affordable therapeutic option against EV71 infection. Full article

(This article belongs to the Special Issue Plant Expression Systems for Bioproduct Production)

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Review

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21 pages, 755 KiB

Open AccessEditor’s ChoiceReview

Development of Systems for the Production of Plant-Derived Biopharmaceuticals

by Ki-Beom Moon, Ji-Sun Park, Youn-Il Park, In-Ja Song, Hyo-Jun Lee, Hye Sun Cho, Jae-Heung Jeon and Hyun-Soon Kim

Plants 2020, 9(1), 30; https://doi.org/10.3390/plants9010030 - 24 Dec 2019

Cited by 70 | Viewed by 15588

Abstract

Over the last several decades, plants have been developed as a platform for the production of useful recombinant proteins due to a number of advantages, including rapid production and scalability, the ability to produce unique glycoforms, and the intrinsic safety of food crops. The expression methods used to produce target proteins are divided into stable and transient systems depending on applications that use whole plants or minimally processed forms. In the early stages of research, stable expression systems were mostly used; however, in recent years, transient expression systems have been preferred. The production of the plant itself, which produces recombinant proteins, is currently divided into two major approaches, open-field cultivation and closed-indoor systems. The latter encompasses such regimes as greenhouses, vertical farming units, cell bioreactors, and hydroponic systems. Various aspects of each system will be discussed in this review, which focuses mainly on practical examples and commercially feasible approaches. Full article

(This article belongs to the Special Issue Plant Expression Systems for Bioproduct Production)

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