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Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors

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Chapter 3
Plant Derived Bioactive Molecules:
Culture Vessels to Bioreactors

Poojadevi Sharma, Sonal Sharma, Sheetal Yadav, Anshu Srivastava,

Indira Purohit, and Neeta Shrivastava

Abstract Bioactive compounds are the compounds having pharmacological or

toxicological effects on humans and animals. At present, the bioactive compounds
are categorized into two groups, secondary metabolites (originating from natural
sources like plants, mammals, fungi, bacteria etc.) and therapeutic recombinant
molecules (which are produced by using recombinant DNA technology in bacteria,
mammals, plants etc.). Of the compounds produced from various sources, second-
ary metabolites produced from plants command highest market demand. Plants are
also proved to be an ideal host system for the production of recombinant therapeutic
molecules. Therefore, there has been a long pursuit for development of a technology
which can provide high yielding plant based bioactive production system. An amal-
gam of plant cell culture and bioreactor technology was crucial in this direction. The
plant bioreactor technology, so developed, has been put to the test many times for
commercial scale production of plant bioactive molecules. There have been
instances of success, but in general growth of plant bioreactor industry has been
very slow. This chapter highlights various aspects of slow but successful growth of
plant based bioactive production from culture vessel to bioreactor. We have evalu-
ated the key drivers and accelerators which have made the journey of plant bioreac-
tor industry successful. Speed breakers of this journey have also been discussed.
Thorough and rigorous analysis of these parameters may help the industry/academia
to speed up the growth of plant bioreactor industry for the production of
bioactives.

Keywords Bioactives • Plant Bioreactors • Culture vessels

P. Sharma • S. Sharma • S. Yadav • A. Srivastava • I. Purohit • N. Shrivastava (*)

Department of Pharmacognosy and Phytochemistry, B. V. Patel
Pharmaceutical Education and Research Development (PERD) Centre,
Sarkhej – Gandhinagar Highway, Thaltej, Ahmedabad, Gujarat 380054, India
e-mail: neetas@perdcentre.com; neetashrivastava@hotmail.com

© Springer Science+Business Media Dordrecht 2014 47

K.-Y. Paek et al. (eds.), Production of Biomass and Bioactive Compounds Using
Bioreactor Technology, DOI 10.1007/978-94-017-9223-3_3
48 P. Sharma et al.

3.1 Plant Bioreactor for Bioactives: Achievements vs

Expectations

Bioactive compounds are the compounds having pharmacological or toxicological

effects on humans and animals. Historic representatives of bioactives are secondary
metabolites obtained from various natural sources like bacteria, fungi, mammals,
plant etc. With the advent of rDNA technology therapeutic recombinant molecules
are also included in this category. Amongst the various natural sources, bioactive
molecules from plants command highest consumer demand. However, the commer-
cial productivity of these molecules is influenced by low yield from natural sources
(bioactive molecules are usually less than 1 % of the plant dry weight) and high cost
of chemical synthesis. Therefore, plant cell culture technology was looked upon as
an alternative production system for these high valued molecules. Subsequently, an
amalgamation of plant cell culture and bioreactor technology has resulted in devel-
opment of successful plant bioreactor technology for commercial scale production
of plant bioactive molecules. Shikonin was the first bioactive to be produced at com-
mercial level, Mitsui Petrochemical Industry Co. Ltd. (Japan) achieved this feat
from plant cells of Lithospermum erythrorhizon in 750 L bioreactor in 1984. After
a dry spell of about 20 years, another success was reported in the year 2002 for the
production of Taxol by Bristol-Myers Squibb and Phyton Biotech, Inc. Till date, it
is the largest commercial application of plant cell culture, utilizing the Chinese yew
(Taxus chinensis) cultivated in 75,000 L bioreactors [1].
Realizing the potential of plant as an apt host system for recombinant therapeutic
protein, the plant bioreactor industry took a leap in this area as well. Year 2012 was
a hallmark year for plant bioreactor industry for recombinant protein production.
The FDA (USA) gave its first ever clearance to a plant-made pharmaceutical prod-
uct, Elelyso™ for treating Gaucher’s disease [2]. A list of plant based bioactives
produced commercially at bioreactor level and their manufacturers is given in
Table 3.1. These examples clearly indicate that plant based bioactive production can
be viably upscaled from culture vessels to bioreactors to achieve market scale pro-
ductivities. However, the timelines of success do hint a fact that growth of plant
bioreactor industry has been slow (Fig. 3.1). This chapter aims at discussion of vari-
ous aspects of slow but successful growth of plant based bioactive production from
culture vessel to bioreactor and also makes plant scientists to ponder over exploiting
the potential of plant cell with measures to take care about its limitations (Fig. 3.2).

3.2 Key Drivers for Success in Plant Bioreactor Technology

Applied facet of plant tissue culture technique has provided a platform for mass pro-
duction of plant derived bioactives using bioreactor technologies. Culture vessel
phase of plant tissue culture technique helps in providing proof of concept which can
further be translated into mass production strategies. Mass production in plant
3 Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors 49

Table 3.1 Plant based bioactives produced commercially at bioreactor level [10, 38]
Plant system Bioactive Manufacturer
Heterologous proteins
Daucus carota Elelyso™ Protalix (Israel)
Oryza sativa Cell culture products Invitria (USA)
(rh-lactoferrin, rh-albumin),
(rh-lyzozyme, rh-transferrin)
Secondary metabolites
Catharanthus roseus Arbtin Mitsui Chemicals, Inc. (Japan)
Coptis japonica Berberines Mitsui Chemicals, Inc. (Japan)
Echinacea purpurea Echinacea polysaccharides Diversa (Germany)
Echinacea angustifolia
Panax ginseng Ginseng Nitto Denko Corporation (Japan)
Taxus spp Paclitaxel Phyton Biotech, Inc. (USA/
Germany) Genexol® – Samyang
Genex (Korea)
Podophyllum Podophyllotoxin Nippon oil (Japan)
Coleus blumei Rosmarinic acid A. Nattermann & Cie.GmbH
(Germany)
Duboisia spp Scopolamine Sumitomo Chemical Co., Ltd.
(Japan)
Lithospermum Shikonin Mitsui Chemicals, Inc.(Japan)
erythrorhizon

Fig. 3.1 Industrial bioreactor based bioactive production and market launch timeline. First mar-
keted secondary metabolite (SM) was Shikonin by Mitsui Petrochemical Industry Co. Ltd in 1984
and second was Paclitaxel by Bristol-Myers Squibb in 2002. The gap was about 20 years. Following
this, many secondary metabolites were launched in market using bioreactor technology. The only
recombinant protein (RP) produced by bioreactor technology is human glucocerebrosidase (human
prGCD) by Protalix BioTherapeutics which was launched recently in 2012
 50

Fig. 3.2 Key drivers responsible for success stories of plant bioactives, accelerators to excel this success and different speed breakers which hamper the
growth. Large scale production of natural as well as recombinant therapeutic molecules can be done by three ways- field cultivation, plant tissue culture and
biomass production in bioreactors. Plant tissue culture can provide platform for production of bioactives for bioreactors. Process can be developed and opti-
mized in culture vessels at small scale and then taken to fermenters. Some allied advance technologies can be applied in vivo to enhance the production
P. Sharma et al.
3 Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors 51

bioreactors provides stable yields in less space and controlled conditions. Following
systems are recognized as key drivers for the success of plant bioreactor technology.

3.2.1 Callus and Cell Suspension Culture

Cell suspension culture provides higher biomass, fast growth and homogeneity in
cells. Various secondary metabolites such as taxol, catharanthine have been pro-
duced using the cell suspension culture and callus culture at laboratory scale [3].
BY-2 and NT-1 cell lines of Tobacco are well characterized to support the produc-
tion of recombinant bioactive molecules [4]. Many proteins including human anti-
rabies monoclonal antibody and erythropoietin have been successfully produced
using these cell lines [5]. Further, these cell lines have potentiality to secrete 0.6–
80 KD protein molecules in the medium which helps in the reduction of down-
stream processing cost [6]. Other plant cells such as rice, tomato and carrot have
also been used for this purpose [7–9].
Suspension cultures have also proven their potential for their immediate applica-
tion in bioreactors for industrial scale production. Elelyso (Taliglucerase alfa), a
May 2012 USFDA approved recombinant drug for the treatment of Gaucher’s
disease and high paclitaxel producing Taxus baccata cells are some other examples
for commercial scale production using cell suspension cultures in bioreactors [2,
10]. Suspension cultures of Glycine max and Nicotiana tabacum at capacity of
10–100 L, suspension culture of Hyoscymus muticus for the production of
hyoscyamine in 100 L capacity bioreactor, production of anthraquinones form the
cultures of Frangula alnus, production of azadirachtin from Azadirachta indica are
some of the culture in the pipeline of commercialization [11–13].
In our laboratory, we have seen the potential of callus culture for the production
of plant bioactive molecule vasicine. We found 0.011 % w/w vasicine concentration
in leaf callus of Adhatoda vasica and 0.13 % w/w concentration of total sennosides
in callus mass of Cassia senna [14].

3.2.2 Hairy Root Culture

Hairy root cultures being differentiated cultures are successful system in the
production of bioactive molecules using bioreactors. Various secondary metabolites
such as berberine, resveratrol and taxol as well as recombinant proteins such as
murine lgG1 monoclonal antibody are successfully produced in this system [5, 15–
17]. Hairy root cultures are genetically stable for long time and provide consistent
expression of proteins in a relatively short time with higher biomass [18]. Hairy root
culture of Panax ginseng is the present successful example for the commercialization
of this technique for biomass production [3]. Reports have demonstrated the success
of Atropin and Ginseng production through hairy roots which is more than the field
52 P. Sharma et al.

grown plants. ROOTec, a German based company produces camptothecin and

podophyllotoxin from the hairy root cultures [19].

3.2.3 Bioreactor Design

Novel design concept has played a pivotal role in success stories of plant bioreac-
tors. Different culture types require different optimized design considerations.
Although many of the bioactives produced successfully in the conventional micro-
bial bioreactors such as stirred tank reactor, bubble column, airlift etc. but the differ-
ent prerequisites of plants such as higher sensitivity to shear stress due to a rigid cell
wall, long generation time etc. raise the need of some modifications. Bioreactors
with good mixing and lower shear stress are preferable for cell cultures [12, 20].
However, the most recent development in this field is the use of disposable bioreac-
tors containing growth chamber made up of FDA approved biocompatible plastics
[12]. These bioreactors use pre-sterilized bags which reduce the possibility of con-
tamination and hence unnecessary efforts and costs required to maintain the produc-
tion safety are negligible. US FDA approved Elelyso (Taliglucerase alfa) was
produced in 400 L capacity disposable bioreactors by Protalix [10]. High paclitaxel
production using Taxus baccata cells is another successful example of Big wave™
disposable bioreactor. Nestle R&D centre in France developed two disposable bio-
reactors of capacity 10–100 L (the wave and undertow bioreactors and slug bubble
bioreactors) for culturing cell suspension of Glycine max and Nicotiana tabacum.
Ebb and flow bioreactor is used to produce Hyoscyamine from the cell suspension
cultures of Hyoscymus muticus of 100 L capacity [12, 13].
The bioreactors used for hairy root cultures are of three types: liquid phase, gas
phase and hybrid [10]. Gas phase nutrient mist bioreactors and temporary immersion
systems save the roots from hyperhydricity [21]. Introduction of meshes, cages and
polyurathene foam for immobilization of roots give opportunity to culture roots in
submerged conventional bioreactors [12]. Bubble column and airlift reactors are
more successful conventional reactors than stirred tank at commercial level for roots
due to their simplicity. However, hyperhydricity is one of the major concerns, so mist
bioreactors or temporary immersion systems are preferred. The hairy root based com-
pany ROOTec developed their own mist bioreactors for the production of many sec-
ondary metabolites from the hairy root cultures of different plant species [19] (http://
www.rootec.com/en/products/all-products). Many high capacity disposable reactors
are also available commercially for the production from hairy roots [12].

3.2.4 Process Design

Operational considerations are also important factors for success of bioreactors based
on cell cultures and hairy root cultures. Designing of bioreactor systems for suspension
cultures is directed towards the reduction shear stress. Proper oxygen supply and
3 Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors 53

gaseous exchange are a very critical factor in case of cell suspension cultures. More
gaseous flow than required can cause evaporation of some essential components.
Improper aeration gives rise to foaming problem in cell culture during scale up.
Temperature should be such which should not affect the bioprocess activities in the
culture with the maintenance of the metabolite production. The optimum temperature
should be 20–23 °C but can vary with the species as well as with the type of product
[20, 22]. Many cell suspension cultures have tendency to form aggregates with differ-
ent morphology. Deviation from the desired type of aggregation may affect the culture
growth and cell-cell interaction which is necessary to maintain the productivity.
Viscosity of culture medium changes due to formation of aggregates and high biomass.
Accumulation of other metabolites can also change the rheology of the medium and
hence growth. Consideration of shear stress is also necessary for the hairy roots as it can
activate the wound response in hairy root cultures resulting in callus formation, thus
causing reduction in productivity [20]. Roots can form a network due to which uniform
supply of the medium may be obstructed. Mineral elements are the very important fac-
tors for the growth of roots and to increase biomass. Immersion time is a critical factor
for root cultures and over lodging can cause hyperhydricity. Inoculum size also affects
the productivity. More tissue mass can cause problems during scaling up.

3.3 Accelerators for the Plant Bioreactor Technology

Use of in vitro culture system coupled with genetic engineering has proved as an
accelerator in the bioreactor technology. Prospects and skills of production of non-
botanical products like expression of human genes in the plant system, transfer of
non-botanical or trans-botanical metabolic pathway specific gene in the culture sys-
tem have proved to be potential accelerators of the technology. Some of the examples
like overexpression of genes encoding Limonene synthase in peppermint, overex-
pression of Chalcone isomerase resulting in increased flavonoids upto 78 % in tomato
peel (Lycopersicum esculentum) [23], expression of a set of genes from marine
sources encoding the fatty acid chain elongation and desaturation enzymes required
for the synthesis of LC-PUFA from their C18 PUFA precursors in Arabidopsis thali-
ana seed, have helped the technology to be commercially more viable [24].

3.4 Speed Breakers in Plant Bioreactor Technology

As evident in Fig. 3.1 the pace of applied and commercialized growth of plant bio-
reactors is very slow in spite of having lot of potential and advantage over other
bioreactor systems. Major reasons of this slow pace are associated with inherent
complicated nature and rich diversity in the plant systems, and some are with the
gap between academics and industries. Limited knowledge/acceptability of genetic
engineering and transgenics are another eclipse in this growth. Following are some
of the troughs which make the growth slow and which should be worked out to get
maximum output by investing time and energies very smartly.
54 P. Sharma et al.

3.4.1 Plant Culture Systems

Many plants are non-amenable to grow in culture, those that could be easily grown
under suitable conditions may lack desired biosynthetic activity or yield is too low
to be commercially viable. Inherent features of plant cell system chosen for up scal-
ing in plant bioreactor itself sometimes limit the process. Plant cell suspension cul-
tures are heterogeneous in nature, therefore, bioactive yield from such cultures is
variable. Probability of genetic instability makes the production inconsistent. In
case of hairy root cultures, their growth creates a tight matrix within a culture, lead-
ing to nutrient transport limitations that result in areas of senescent cells [25]. These
nutrient gradients also attribute to variability in root growth and productivity. Yield
can also impede due to hyperhydricity which makes the roots very vulnerable.
Further, both plant cell suspensions and hairy root cultures are sensitive to shear
stress and mechanical agitation resulting in cell wounding [25]. Low protein pro-
duction, contamination, high downstream processing cost, degradation of secreted
protein in medium by proteolytic enzymes are the major limitations for the produc-
tion of recombinant bioactive molecules in plant systems.

3.4.2 Post Translational Modifications

Being safer for humans, plants are considered as the most suitable host system for
engineered biothearpeutics. Plants are easily approachable and can be utilized in
raw form, this has given birth to the concept of edible vaccines. However, the highly
modified and evolved internal physiological system of plants works as a barrier to
produce foreign proteins in their original efficient form. There are many events such
as proteolysis, misfolding, aggregation, oxidation of methionine, deamination of
asparagine and glutamine, and glycosylation occurring in cells during the post
translational modifications (PTMs) of recombinant bioactive molecules. All of
these PTMs are important for the stability, solubility, bioactivity, efficacy and effi-
cient secretion of the proteins. Glycosylation is one of the most crucial steps regard-
ing the addition of correct glycans to the proteins in cells. Plants are highly
competent and capable for the production of large and complex proteins and addi-
tionally plants provide higher biomass which is directly linked to the maximum
production of bioactive molecules. However, during the post translational modifica-
tions plants add some unwanted β (1,2)-Xylose and core α (1,3)-Fucose residues
instead of core α (1,6)-Fucose residues and terminal Neuraminic acid (NeuAc)
resulting it to become immunogenic to humans [26, 27]. NeuAc, which is responsi-
ble for the activity and stability of the proteins, is not found in a plant. Till date, many
bioactive molecules such as erythropoietin, human haemoglobin, human epidermal
growth factors and human serum albumin etc. have been produced in plants [28].
Still, none of these products are available in market except glucocerebrosidase pro-
duced in the carrot suspension cells [8]. Therefore, protein glycosylation is one of
3 Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors 55

the major concern in the plant cells for the production of bioactive molecules which
needs proper attention to resolve the problem of improper glycosylation in plants.

3.4.3 Bioreactor System Designs

The concept of bioreactors originated mainly for the microbial products and then
extended to the higher eukaryotic systems. Earlier the microbial bioreactor designs
were utilized for plants but different requirements of the systems raise the need of
designs specifically developed for the plants like bubble column bioreactor, tempo-
rary immersion systems, continuous flow reactor, etc. An important critical step in
general for all bioreactors is determination of inoculum size for differentiated plant
in vitro systems [29]. Achieving uniform distribution of the cultures in the growth
chamber is very difficult. Different techniques need to be used for inoculation in a
bioreactor like using of a seed vessel to obtain inoculum roots that were transferred
aseptically by means of helical screw in a 500-L hybrid reactor; homogenization and
transfer of the biomass into bioreactors as slurry; change of the cultivation mode
from batch to fed-batch [30]. Different bioreactor types can significantly affect the
culture growth and product accumulation. Therefore, development of such processes
needs optimization in direction to develop plant bioreactor based technologies.

3.4.4 Scientific Mindset

Scientists focus primarily on research rather than business or regulatory aspects.

Basic researches are driven mainly by the curiosity and interest of a particular sci-
entist. Research areas like development of in vitro cultures, elicitation, plant genetic
engineering, bioreactor upscaling, etc. requires long period of time to get success-
fully translated into commercial value. Scientists are more comfortable with their
slow pace of research and have patience to carry on research for years even with
negative results because curiosity to solve the unresolved mystery. Such slow and
long time frame for a research solution cannot cope up with the fast changing indus-
try market interests. Academia should make to stand stable to cope up with fast
changing technologies. Lack of proper resources and infrastructure also inhibit the
scientists to take their efforts to commercially practical scale.

3.4.5 Industrial Mindset

Establishing plant fermentation systems involve large capital start-up costs. The
batch times for plant cultures are very large, so maintenance and monitoring needs
are also big. Industry thinks in terms of short range goals and wants result in short
expected time frames. Industry cannot risk delays and loss of profits. While
56 P. Sharma et al.

considering money investment, industry prefers a low risk industry with proved
profitable products. Mammalian cells are widely used by industry for the produc-
tion of recombinant therapeutics which exhibit satisfactory glycosylation. Further
industry is quite experienced with handling regulatory guidelines and cGMP issues
for mammalian bioreactor industry as compared to plant bioreactor industry which
is still emerging. On account of these factors, industry is still hesitant to venture into
commercial plant bioreactor sector.

3.4.6 Lack of Academic and Industry Synergism

Differences in mindsets of an academician and a corporate person stop them to come

together on a single platform. The working culture differences keep these two heads
apart. An industrialist always thinks for profit and prefers to secure their researches in
the form of patents or in the name of trade secrets. In contrast to institutes, leakage and
sharing of knowledge is unacceptable in an industry. Demands of huge returns limit
the institutes to work with industries. Mutually exclusive preferences, demands,
visions and research achievements criteria weaken the faculty and firm collaboration.
Slow pace of research in the academic institutes does not match with the higher expec-
tations of industries of getting quick results. Examples are there which exhibited the
power of crossing the lines to work together and enhances the surety of success. The
successful production of shiknonin from the plant culture was a result of combined
efforts of Kayoto University and Mitusi petrochemical in Japan. Another example of
successful alliance is Kitasato University and Nitto Denko in Japan, resulting in the
production of ginseng [31]. Existence of very few illustrations shows the necessity for
more synergistic efforts with crystalline purity, sincere, honesty and immaculate trans-
parency which are very essential to create and also to maintain the collaborations.

3.4.7 Public Mindset

The public i.e. consumers are the ultimate decision makers for a product to be com-
mercially successful in market. At present, public has many issues with plant
recombinant bioactive molecules.

Ethical Issues

Ethical issues of certain groups of public, including religious bodies have been
major determinants in withdrawal of many such bioactive molecule producing
plants from fields. They find it unethical or inhumane to introduce gene of animal or
human origin into plants [32, 33].
3 Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors 57

Health Issues

General public is also worried about risk of allergenicity (usually glycoproteins) of

these new recombinant bioactives from plants [34]. Transgenic technologies use
sensitive genes such as, antibiotic marker genes and promoter sequences derived
from viruses. During repetitive plant transformations, antibiotic resistance genes
will accumulate and plant breeders will soon encounter difficulties in locating new,
harmless antibiotic marker genes. The obvious fear is that antibiotic marker genes
could be recruited into humans along with the gene for foreign proteins (and domes-
tic animals) rendering antibiotics ineffective in curing bacterial infections. Plants
producing insulin, growth hormones, plantibodies etc. are direct applications of
genetic engineering for human health but for strict vegetarians it could pose an ethi-
cal issue.

Socio-economic Issues

Developing countries believe that genetically modified whole plant bioreactors are
profit crops majorly for western developed countries and developing countries are
only being exploited in the business for growing these plants at cheap rates. In midst
of such anti-GM public perception, it is not easy to convince investors to fund plant
bioreactor industry or even any related academic research.

3.4.8 Environmental Risks from Transgenics

Plants prove their potential to be a good choice for foreign protein production
hence they are themselves considered as a natural bioreactors as genetically modi-
fied crops but these efficient bioreactors can harm ecosystem, food webs, biodiver-
sity and germplasm. There are ample evidences that transgenic crops and their
genes, through pollen dispersal, can spread even between species. There is also a
low probability of chloroplast movement from transgenic oilseed rape into wild
species [35]. The effects of transgene escape on the environment are uncertain,
result into “genetic pollution”, for example tailoring herbicide resistance, poses
threat that what and how much of the herbicide should be used, its persistence and
residual effects and development of resistant target species or gene flow to non-
target species. This can also threat ecosystems and biodiversity. Gene flow
increases outcrossing that out competes in the ecosystem. Genetic transformation
can harm biodiversity by reduction in insects that serve as food at higher trophic
levels. For instance monarch butterflies feeding on GM corn leaves had deduced
growth [36]. These risks may not be visible instantaneously but should be taken
care of.
58 P. Sharma et al.

3.4.9 Regulatory Concerns/IPR Conflicts

The commercialization of plant biotechnology has advanced rapidly over the past
5 years. Intellectual property rights, mainly in the form of patents, have been funda-
mental to the commercial development of the technology. Several hundred patents on
plant genes, techniques for genetic modification and transgenic plants have now been
granted and many more have been filed. Although patenting in biotechnology gener-
ally is now widely practiced by public and private sector researchers alike, exces-
sively broad claims and restrictive licensing remain a potential threat for innovation.
Patenting and licensing in this area restricts competition and increases monopolies on
key plant technologies. This may further restrict innovation, fair access and trade. The
ultimate outcome in this direction could be decline in willingness to invest in research
and development and share knowledge in public domain. Ownership of genes and the
need for patents is a further area for ethical debate. Innumerable IPR court cases are
filed among farmers and public sectors of developing countries, plant bioreactor
industry, anti-GM NGOs, academicians who developed the technology etc.
Patents based on the natural therapeutic products are also a matter for concern.
Most of the biodiversity is concentrated to some developing countries. They have
ample resources to generate plant based natural products but are deprived in research
resources. Most of the plant based therapeutics extract from the traditional knowl-
edge acquired by the indigenous communities since time immemorial. Traditional
knowledge of these countries is exploited by the multinational companies, modi-
fied, utilized and filed as patents. Resource limitations in developing countries
restrict them to use their legacy for themselves and bound to buy their modified
versions in high amount. Many examples of biopiracy are there like use of turmeric,
neem, hoodia plant, banana extract; melon extract etc. for treating various diseases
provokes the governments to raise voice to protect their traditional knowledge.
Claiming rights related to improvements in plant traits (like enhanced yield of sec-
ondary metabolite) using advance technologies can also raise clashes because the
basic genetic information used for transgenic plant bioreactor development is
extracted from the ecosystems of developing nations [37]. These unnecessary IPR
conflicts slow the research pace and reduce the productivity.
It is clearly evident that neither academia nor even industry would want to
involve in such legal issues, therefore the industry would more likely feel comfort-
able in investing for a proposed plant bioreactor strategy only after thorough thought
process which would require some considerable time investment.

3.5 Conclusion and Future Perspectives

Advances in the biotechnology particularly methods for culturing plant cell cultures
has provided new means for the commercial production of even the rare medicinal
plants and chemical they provide, so there has been a considerable interest in plant
cell cultures as the potential alternative to the traditional agriculture for the
3 Plant Derived Bioactive Molecules: Culture Vessels to Bioreactors 59

industrial production of the secondary metabolites. The objectives of many indus-

tries are to develop plant cell culture techniques to the stage where they yield sec-
ondary products more economically than the whole plant grown under natural
conditions or synthesizing the product.
Design of a suitable bioreactor with low-shear impeller, and selection of an
appropriate mode of cultivation is required for increased metabolite production.
Optimization of medium ingredients by statistical techniques, application of appro-
priate mathematical models for optimized cell cultivation, feeding strategy of meta-
bolic precursors, and extraction of intracellular metabolites by organic solvents can
lead to significant enhancement in productivity of secondary metabolites.

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