ijms-logo

Journal Browser

Journal Browser

Plant-Derived Pharmaceuticals by Molecular Farming 2016

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Bioactives and Nutraceuticals".

Deadline for manuscript submissions: closed (30 August 2016) | Viewed by 87041

Special Issue Editor

Special Issue Information

Dear Colleagues,

Molecular farming (MF), in terms of production scale and economy, product safety, ease of storage, and distribution, is superior to any other current commercial alternative. Despite MF’s promising benefits, the commercialization of MF-derived products is lagging behind due to uncertainty and safety issues. A particularly troubling issue concerns the adaptation of good manufacturing practice regulations for transgenic organisms. Although MF is not yet routinely commercialized, interest and investment in MF are extending rapidly. Ultimately, MF will progress and become a core commercial development in biotechnology.

This Special Issue aims to provide high quality research concerning a novel gene expression in target organisms, such as green algae or plants, and the utilization of this technique to manufacture pharmaceutical proteins. Possible MF products include biopharmaceuticals, nutraceuticals, vaccines, metabolites, and foreign proteins for industrial purposes. Possible topics cover research in MF’s industrial and preclinical/clinical applications, or MF’s environmental impacts. I wish to thank all authors for their contributions to this Special Issue.

Chang Won Choi
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • molecular farming
  • transgenic organisms and/or transient expression
  • industrial, preclinical/clinical applications
  • environmental impacts
  • algae, plants, biopharmaceuticals, nutraceuticals, vaccines, metabolites, foreign proteins for industrial purposes

Published Papers (10 papers)

Order results
Result details
Select all
Export citation of selected articles as:

Research

Jump to: Review

2941 KiB  
Article
Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana
by Kalimuthu Karuppanan, Sifti Duhra-Gill, Muchena J. Kailemia, My L. Phu, Carlito B. Lebrilla, Abhaya M. Dandekar, Raymond L. Rodriguez, Somen Nandi and Karen A. McDonald
Int. J. Mol. Sci. 2017, 18(1), 89; https://doi.org/10.3390/ijms18010089 - 04 Jan 2017
Cited by 6 | Viewed by 6208
Abstract
Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused [...] Read more.
Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The N-glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of N-glycans consists of complex type structures in both protein samples. The most abundant (>50%) N-glycan structure was GlcNAc2(Xyl)Man3(Fuc)GlcNAc2 in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose N-glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in Nicotiana benthamiana plants with apoplast targeting. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Figure 1

6533 KiB  
Article
Direct LAMP Assay without Prior DNA Purification for Sex Determination of Papaya
by Chi-Chu Tsai, Huei-Chuan Shih, Ya-Zhu Ko, Ren-Huang Wang, Shu-Ju Li and Yu-Chung Chiang
Int. J. Mol. Sci. 2016, 17(10), 1630; https://doi.org/10.3390/ijms17101630 - 24 Sep 2016
Cited by 8 | Viewed by 6627
Abstract
Papaya (Carica papaya L.) is an economically important tropical fruit tree with hermaphrodite, male and female sex types. Hermaphroditic plants are the major type used for papaya production because their fruits have more commercial advantages than those of female plants. Sex determination [...] Read more.
Papaya (Carica papaya L.) is an economically important tropical fruit tree with hermaphrodite, male and female sex types. Hermaphroditic plants are the major type used for papaya production because their fruits have more commercial advantages than those of female plants. Sex determination of the seedlings, or during the early growth stages, is very important for the papaya seedling industry. Thus far, the only method for determining the sex type of a papaya at the seedling stage has been DNA analysis. In this study, a molecular technique—based on DNA analysis—was developed for detecting male-hermaphrodite-specific markers to examine the papaya’s sex type. This method is based on the loop-mediated isothermal amplification (LAMP) and does not require prior DNA purification. The results show that the method is an easy, efficient, and inexpensive way to determine a papaya’s sex. This is the first report on the LAMP assay, using intact plant materials-without DNA purification-as samples for the analysis of sex determination of papaya. We found that using high-efficiency DNA polymerase was essential for successful DNA amplification, using trace intact plant material as a template DNA source. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

3778 KiB  
Article
New Natural Pigment Fraction Isolated from Saw Palmetto: Potential for Adjuvant Therapy of Hepatocellular Carcinoma
by Hor-Yue Tan, Ning Wang, Masao Takahashi, Yigang Feng, Hongyun Li and Yibin Feng
Int. J. Mol. Sci. 2016, 17(8), 1277; https://doi.org/10.3390/ijms17081277 - 05 Aug 2016
Cited by 3 | Viewed by 4931
Abstract
For the first time, we discovered a small proportion of aqueous fraction from Saw Palmetto apart from the fatty acid-rich fraction exhibited pharmacological activity. Therefore, this study aims to explore the anti-tumor potential of red pigmented aqueous fraction of Saw Palmetto, NYG on [...] Read more.
For the first time, we discovered a small proportion of aqueous fraction from Saw Palmetto apart from the fatty acid-rich fraction exhibited pharmacological activity. Therefore, this study aims to explore the anti-tumor potential of red pigmented aqueous fraction of Saw Palmetto, NYG on human hepatocellular carcinoma and its possible targets. Subcutaneous xenograft and orthotopic implantation models of HCC were used to evaluate the tumor inhibitory effect of NYG. Human hepatocellular carcinoma (HCC) cell lines and human umbilical vein endothelial cells (HUVEC) were used as in vitro model. The mRNA expression was conducted by qPCR. Protein expression was monitored by immunoblotting and immunohistochemistry. Cell migration and blood vessel formation were determined by chamber assay and tube formation assay, respectively. Significant tumor inhibition of NYG in dose-dependent manner was observed on subcutaneous xenograft and orthotopic HCC model. NYG has no direct action on cell viability or VEGF secretion of HCC cells. However, NYG reduced in vitro migration and vessel formation activities of HUVEC cells, as well as in vivo intratumoral neovascularization. NYG attenuated extracellular signal-regulated kinases (ERK) activation in endothelial cells, which may be associated with the suppression of migration and tube formation of HUVEC. NYG suppressed tumor expansion of HCC via inhibiting neovascularization, and may be potential adjuvant treatment for HCC. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

4779 KiB  
Article
Up-Regulation of PAI-1 and Down-Regulation of uPA Are Involved in Suppression of Invasiveness and Motility of Hepatocellular Carcinoma Cells by a Natural Compound Berberine
by Xuanbin Wang, Ning Wang, Hongliang Li, Ming Liu, Fengjun Cao, Xianjun Yu, Jingxuan Zhang, Yan Tan, Longchao Xiang and Yibin Feng
Int. J. Mol. Sci. 2016, 17(4), 577; https://doi.org/10.3390/ijms17040577 - 16 Apr 2016
Cited by 35 | Viewed by 7013
Abstract
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death and its prognosis remains poor due to the high risk of tumor recurrence and metastasis. Berberine (BBR) is a natural compound derived from some medicinal plants, and accumulating evidence has shown its [...] Read more.
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death and its prognosis remains poor due to the high risk of tumor recurrence and metastasis. Berberine (BBR) is a natural compound derived from some medicinal plants, and accumulating evidence has shown its potent anti-tumor activity with diverse action on tumor cells, including inducing cancer cell death and blocking cell cycle and migration. Molecular targets of berberine involved in its inhibitory effect on the invasiveness remains not yet clear. In this study, we identified that berberine exhibits a potent inhibition on the invasion and migration of HCC cells. This was accompanied by a dose-dependent down-regulation of expression of Cyclooxygenase-2 (COX-2), nuclear factor kappa B (NF-κB), urokinase-type plasminogen activator (uPA) and matrix metalloproteinase (MMP)-9 in berberine-treated HCC cells. Furthermore, berberine inactivated p38 and Erk1/2 signaling pathway in HCC cells. Primarily, this may be attributed to the up-regulation of plasminogen activator inhibitor-1 (PAI-1), a tumor suppressor that can antagonize uPA receptor and down-regulation of uPA. Blockade of uPA receptor-associated pathways leads to reduced invasiveness and motility of berberine-treated HCC cells. In conclusion, our findings identified for the first time that inactivation of uPA receptor by up-regulation of PAI-1 and down-regulation of uPA is involved in the inhibitory effect of berberine on HCC cell invasion and migration. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

7823 KiB  
Article
Chemical Elicitor-Induced Modulation of Antioxidant Metabolism and Enhancement of Secondary Metabolite Accumulation in Cell Suspension Cultures of Scrophularia kakudensis Franch
by Abinaya Manivannan, Prabhakaran Soundararajan, Yoo Gyeong Park and Byoung Ryong Jeong
Int. J. Mol. Sci. 2016, 17(3), 399; https://doi.org/10.3390/ijms17030399 - 18 Mar 2016
Cited by 44 | Viewed by 7740
Abstract
Scrophularia kakudensis is an important medicinal plant with pharmaceutically valuable secondary metabolites. To develop a sustainable source of naturaceuticals with vital therapeutic importance, a cell suspension culture was established in S. kakudensis for the first time. Friable calli were induced from the leaf [...] Read more.
Scrophularia kakudensis is an important medicinal plant with pharmaceutically valuable secondary metabolites. To develop a sustainable source of naturaceuticals with vital therapeutic importance, a cell suspension culture was established in S. kakudensis for the first time. Friable calli were induced from the leaf explants cultured on a Murashige and Skoog (MS) medium containing 3.0 mg·L−1 6-benzyladenine (BA) in a combination with 2 mg·L−1 2,4-dichlorophenoxy acetic acid (2,4-D). From the callus cultures, a cell suspension culture was initiated and the cellular differentiation was investigated. In addition, the effect of biotic elicitors such as methyl jasmonate (MeJa), salicylic acid (SA), and sodium nitroprusside (SNP) on the accumulation of secondary metabolites and antioxidant properties was demonstrated. Among the elicitors, the MeJa elicited the accumulation of total phenols, flavonoids, and acacetin, a flavonoid compound with multiple pharmaceutical values. Similarly, the higher concentrations of the MeJa significantly modulated the activities of antioxidant enzymes and enhanced the scavenging potentials of free radicals of cell suspension extracts. Overall, the outcomes of this study can be utilized for the large scale production of pharmaceutically important secondary metabolites from S. kakudensis through cell suspension cultures. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

2024 KiB  
Article
Prenylated Flavonoids from Cudrania tricuspidata Suppress Lipopolysaccharide-Induced Neuroinflammatory Activities in BV2 Microglial Cells
by Dong-Cheol Kim, Chi-Su Yoon, Tran Hong Quang, Wonmin Ko, Jong-Su Kim, Hyuncheol Oh and Youn-Chul Kim
Int. J. Mol. Sci. 2016, 17(2), 255; https://doi.org/10.3390/ijms17020255 - 19 Feb 2016
Cited by 28 | Viewed by 5952
Abstract
In Korea and China, Cudrania tricuspidata Bureau (Moraceae) is an important traditional medicinal plant used to treat lumbago, hemoptysis, and contusions. The C. tricuspidata methanol extract suppressed both production of NO and PGE2 in BV2 microglial cells. Cudraflavanone D (1), isolated from [...] Read more.
In Korea and China, Cudrania tricuspidata Bureau (Moraceae) is an important traditional medicinal plant used to treat lumbago, hemoptysis, and contusions. The C. tricuspidata methanol extract suppressed both production of NO and PGE2 in BV2 microglial cells. Cudraflavanone D (1), isolated from this extract, remarkably suppressed the protein expression of inducible NO synthase and cyclooxygenase-2, and decreased the levels of NO and PGE2 in BV2 microglial cells exposed to lipopolysaccharide. Cudraflavanone D (1) also decreased IL-6, TNF-α, IL-12, and IL-1β production, blocked nuclear translocation of NF-κB heterodimers (p50 and p65) by interrupting the degradation and phosphorylation of inhibitor of IκB-α, and inhibited NF-κB binding. In addition, cudraflavanone D (1) suppressed the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 MAPK pathways. This study indicated that cudraflavanone D (1) can be a potential drug candidate for the cure of neuroinflammation. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

1558 KiB  
Article
Cissus sicyoides: Pharmacological Mechanisms Involved in the Anti-Inflammatory and Antidiarrheal Activities
by Fernando Pereira Beserra, Raquel De Cássia Santos, Larissa Lucena Périco, Vinicius Peixoto Rodrigues, Luiz Ricardo De Almeida Kiguti, Luiz Leonardo Saldanha, André Sampaio Pupo, Lúcia Regina Machado Da Rocha, Anne Lígia Dokkedal, Wagner Vilegas and Clélia Akiko Hiruma-Lima
Int. J. Mol. Sci. 2016, 17(2), 149; https://doi.org/10.3390/ijms17020149 - 23 Jan 2016
Cited by 14 | Viewed by 6050
Abstract
The objective of this study was to evaluate the pharmacological mechanisms involved in anti-inflammatory and antidiarrheal actions of hydroalcoholic extract obtained from the leaves of Cissus sicyoides (HECS). The anti-inflammatory effect was evaluated by oral administration of HECS against acute model of edema [...] Read more.
The objective of this study was to evaluate the pharmacological mechanisms involved in anti-inflammatory and antidiarrheal actions of hydroalcoholic extract obtained from the leaves of Cissus sicyoides (HECS). The anti-inflammatory effect was evaluated by oral administration of HECS against acute model of edema induced by xylene, and the mechanisms of action were analysed by involvement of arachidonic acid (AA) and prostaglandin E2 (PGE2). The antidiarrheal effect of HECS was observed and we analyzed the motility and accumulation of intestinal fluid. We also analyzed the antidiarrheal mechanisms of action of HECS by evaluating the role of the opioid receptor, α2 adrenergic receptor, muscarinic receptor, nitric oxide (NO) and PGE2. The oral administration of HECS inhibited the edema induced by xylene and AA and was also able to significantly decrease the levels of PGE2. The extract also exhibited significant anti-diarrheal activity by reducing motility and intestinal fluid accumulation. This extract significantly reduced intestinal transit stimulated by muscarinic agonist and intestinal secretion induced by PGE2. Our data demonstrate that the mechanism of action involved in the anti-inflammatory effect of HECS is related to PGE2. The antidiarrheal effect of this extract may be mediated by inhibition of contraction by acting on the intestinal smooth muscle and/or intestinal transit. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

Review

Jump to: Research

318 KiB  
Review
The Last Ten Years of Advancements in Plant-Derived Recombinant Vaccines against Hepatitis B
by Young Hee Joung, Se Hee Park, Ki-Beom Moon, Jae-Heung Jeon, Hye-Sun Cho and Hyun-Soon Kim
Int. J. Mol. Sci. 2016, 17(10), 1715; https://doi.org/10.3390/ijms17101715 - 13 Oct 2016
Cited by 21 | Viewed by 9429
Abstract
Disease prevention through vaccination is considered to be the greatest contribution to public health over the past century. Every year more than 100 million children are vaccinated with the standard World Health Organization (WHO)-recommended vaccines including hepatitis B (HepB). HepB is the most [...] Read more.
Disease prevention through vaccination is considered to be the greatest contribution to public health over the past century. Every year more than 100 million children are vaccinated with the standard World Health Organization (WHO)-recommended vaccines including hepatitis B (HepB). HepB is the most serious type of liver infection caused by the hepatitis B virus (HBV), however, it can be prevented by currently available recombinant vaccine, which has an excellent record of safety and effectiveness. To date, recombinant vaccines are produced in many systems of bacteria, yeast, insect, and mammalian and plant cells. Among these platforms, the use of plant cells has received considerable attention in terms of intrinsic safety, scalability, and appropriate modification of target proteins. Research groups worldwide have attempted to develop more efficacious plant-derived vaccines for over 30 diseases, most frequently HepB and influenza. More inspiring, approximately 12 plant-made antigens have already been tested in clinical trials, with successful outcomes. In this study, the latest information from the last 10 years on plant-derived antigens, especially hepatitis B surface antigen, approaches are reviewed and breakthroughs regarding the weak points are also discussed. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

294 KiB  
Review
The Potential for Microalgae as Bioreactors to Produce Pharmaceuticals
by Na Yan, Chengming Fan, Yuhong Chen and Zanmin Hu
Int. J. Mol. Sci. 2016, 17(6), 962; https://doi.org/10.3390/ijms17060962 - 17 Jun 2016
Cited by 153 | Viewed by 13504
Abstract
As photosynthetic organisms, microalgae can efficiently convert solar energy into biomass. Microalgae are currently used as an important source of valuable natural biologically active molecules, such as carotenoids, chlorophyll, long-chain polyunsaturated fatty acids, phycobiliproteins, carotenoids and enzymes. Significant advances have been achieved in [...] Read more.
As photosynthetic organisms, microalgae can efficiently convert solar energy into biomass. Microalgae are currently used as an important source of valuable natural biologically active molecules, such as carotenoids, chlorophyll, long-chain polyunsaturated fatty acids, phycobiliproteins, carotenoids and enzymes. Significant advances have been achieved in microalgae biotechnology over the last decade, and the use of microalgae as bioreactors for expressing recombinant proteins is receiving increased interest. Compared with the bioreactor systems that are currently in use, microalgae may be an attractive alternative for the production of pharmaceuticals, recombinant proteins and other valuable products. Products synthesized via the genetic engineering of microalgae include vaccines, antibodies, enzymes, blood-clotting factors, immune regulators, growth factors, hormones, and other valuable products, such as the anticancer agent Taxol. In this paper, we briefly compare the currently used bioreactor systems, summarize the progress in genetic engineering of microalgae, and discuss the potential for microalgae as bioreactors to produce pharmaceuticals. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

932 KiB  
Review
Plants as Factories for Human Pharmaceuticals: Applications and Challenges
by Jian Yao, Yunqi Weng, Alexia Dickey and Kevin Yueju Wang
Int. J. Mol. Sci. 2015, 16(12), 28549-28565; https://doi.org/10.3390/ijms161226122 - 02 Dec 2015
Cited by 191 | Viewed by 18865
Abstract
Plant molecular farming (PMF), defined as the practice of using plants to produce human therapeutic proteins, has received worldwide interest. PMF has grown and advanced considerably over the past two decades. A number of therapeutic proteins have been produced in plants, some of [...] Read more.
Plant molecular farming (PMF), defined as the practice of using plants to produce human therapeutic proteins, has received worldwide interest. PMF has grown and advanced considerably over the past two decades. A number of therapeutic proteins have been produced in plants, some of which have been through pre-clinical or clinical trials and are close to commercialization. Plants have the potential to mass-produce pharmaceutical products with less cost than traditional methods. Tobacco-derived antibodies have been tested and used to combat the Ebola outbreak in Africa. Genetically engineered immunoadhesin (DPP4-Fc) produced in green plants has been shown to be able to bind to MERS-CoV (Middle East Respiratory Syndrome), preventing the virus from infecting lung cells. Biosafety concerns (such as pollen contamination and immunogenicity of plant-specific glycans) and costly downstream extraction and purification requirements, however, have hampered PMF production from moving from the laboratory to industrial application. In this review, the challenges and opportunities of PMF are discussed. Topics addressed include; transformation and expression systems, plant bioreactors, safety concerns, and various opportunities to produce topical applications and health supplements. Full article
(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)
Show Figures

Graphical abstract

Back to TopTop