A Review on Bioreactor Technology Assisted Plant ...

_____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected];

Asian Journal of Biotechnology and Bioresource Technology 5(3): 1-13, 2019; Article no.AJB2T.51795 ISSN: 2457-0125

A Review on Bioreactor Technology Assisted Plant Suspension Culture

Renugaa Su1, M. Sujarani1, P. Shalini1 and N. Prabhu1*

1Department of Biotechnology, Vivekanandha College of Engineering for Women, Tiruchengode,

Namakkal-637 205, Tamilnadu, India.

Authors’ contributions

This work was carried out in collaboration among all authors. Authors RS, MS and PS designed the study, performed the statistical analysis and wrote the first draft of the manuscript. Author NP

managed the analyses of the study and literature searches. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/AJB2T/2019/v5i330062

Editor(s): (1) Dr. M. M. Youssef, Retired Professor, Food Science and Technology, Faculty of Agriculture, Alexandria University,

Alexandria, Egypt. Reviewers:

(1) Kartika Rathore, Jai Narain Vyas University, India. (2) Hamdy M. Abdel-Rahman, National Research Centre, Egypt.

(3) Mostafa M. Abo Elsoud, National Research Centre, Egypt. (4) T. Gajendran, Anna University, India.

Complete Peer review History: https://sdiarticle4.com/review-history/51795

Received 02 August 2019 Accepted 07 October 2019

Published 17 October 2019

ABSTRACT

This review is related to bioreactors for plant suspension culture and its products. Bioreactor plays an important role in bioprocess engineering. The core of bioprocessing technology is the bioreactor. A bioreactor is basically a device in which the organisms are cultivated and helps in production of desired products in a contained environment. Bioreactors are usually a containment which provides optimal condition for microorganisms in order to produce desired products. In this review, the bioreactor’s principle, working and its types are discussed. Enclosed by unit operations that carry out physical changes for medium preparation and recovery of products, the reactor is where the major chemical and biochemical transformations occur. In many bioprocess, characteristic of the reaction determined to a large extent the economic feasibility of the project. The integration of biosynthesis and separation is considered as a possible approach towards more efficient plant cell and tissue culture. In this review article, the aspects of bioprocess engineering for plant suspension culture and its products, bioreactor types, optimized strategies for production of secondary metabolites also and its industrial applications.

Review Article

Su et al.; AJB2T, 5(3): 1-13, 2019; Article no.AJB2T.51795

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Keywords: Bioreactor; secondary metabolite production; industrial applications. 1. INTRODUCTION Plant tissue culture is generally refers to the in vitro cultivation of any plant segment, that may be a single cell, a tissue or an organ. They are five main types of plant tissue cultures: 1) seed lines of plants, 2) Isolated embryo,3) Isolated plant organs,4) Explant cultures, 5) Isolate cells or small aggregates dispersed in liquid media. Generally, commercial interests are secondary metabolites, which is categorized into essential oils, glycosides alkaloids. These are synthesized by means of bioprocess engineering using bioreactors [1]. Plants have evolved a vast chemical copiousness determined primarily by the requirement to protect themselves from microbial pathogens, Insects and pests, grazing animals. Special bioreactors are developed for some application [2]. Typical bioreactors for plant cell and tissue cultures have been made of glass or stainless steel for more than 40 years. In this area, stirred reactors, rotating drum reactors, airlift reactors, bubble columns, fluidized bed reactors, packed bed reactors and trickle bed reactors with culture volumes up to 75 m

3 as well

as their modifications are the most commonly used bioreactor types in research and commercial production processes [3]. Bioreactors have been developed with disposable manner. These bioreactors consist of a sterile plastic chamber that is partially filled with media (10% to 50%), inoculated with cells and discarded after harvest. The single use chamber eliminating any need for cleaning or sterilization is made of FDA approved biocompatible plastics such as polyethylene, polystyrene and polypropylene. Generally, the disposable bioreactors are low cost, simple to operate and undertaking high process security. It is suggested that their use could improve process efficiency and results by reducing the time to time market of new products. The objective of this review is to critically outline the potential of the bioreactors for secondary metabolite production from suspension cultures, hairy roots and embryogenic cultures [4]. With respect to the types of bioreactors reported in the literature, their classification, application and characterization, here we describe the features of bioreactors as well as summaries the results of hydrodynamic studies (characterization of fluid flow, estimation of mixing time, distribution time, energy input) and investigations of oxygen transport efficiency [5].

2. PLANT CELL CULTURE

There are different plant cell culture techniques, used in industries and research purpose. Majorly, there are different culture types of plant cell culture. They are, Callus, plant cell suspension, Hairy root cultures, Embryogenic and shoot cultures [6].

2.1 Callus Culture Callus is defined as a developing mass of unorganized parenchyma cells, the formation of callus normally is achieved by placing the explant on an appropriate solid growth medium with the necessary components such as plant growth regulators in a Petri dish and incubated. Callus cultures have a developmental stage of cells described as dedifferentiated cells.

2.2 Hairy Roots Hairy roots are generated by the transformation of plants or explants with agro pine and Mann opine type strains of Agrobacterium rhizogenic [7].

2.3 Embryogenic and Shoot Culture

It is used for micro propagation and plant breeding and like hairy roots. They belong to the group of differentiated organ culture. Meristem, seed germination or embryonic culture is used to stable this kind of cultures.

2.4 Plant Cell Suspension Plant cell suspension are highly variable in terms of morphology, rheological characteristics ,growth and metabolic pattern .Rheological characters are important in the process of scale up ,Plant cell suspensions are non-Newtonian fluids ,so the density of this cultures rises proportional to cell density. As a result, plant suspension cells very rarely grow as single cells, they form aggregates based on cell adhesion and results from the segregation of wall extra cellular polysaccharides, this prevents cell separation [8].

3. CULTURE CONDITIONS Mostly of the plant cell cultures requires similar culture conditions as temperature, between 25 and 27ºC, a medium pH of 5.0 and 6.0. Also,


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culture cells need some aeration; this aeration is extremely lower than microbial system. Some cultures have periodic dark/light cycle of 8 hr and 16 hr or continuous introduction of light (0.6 -10 Klux), mammalian cells have to be cultivated in dark. Long lag phases (nearly 120 hours) can be eliminated by plant cell cultures can be initiated with high cell concentrations (10% of culture volume) [9].

3.1 Media

Culturing plant cells are based on nutrient supply. Doubled distilled and deionized water which represents 95% of the media are the main substances used for culture. All culture media have a basal medium with carbon source. Carbon sources include fructose, sucrose, glucose and sorbitol, with organic and inorganic supplements. Supplements such as amino acids, vitamins and cofactors as tocopherol are used for BY-2 sigmoid growth source. The inorganic supplements are macro and microelements, the microelements are in µM concentrations and microelements in mm.

In addition to this nutrient supply needs some phytohormones as growth regulators like auxins, cytokines and gibberellins. Growth regulators affect the growth process, cytokinin which promotes cell division and auxins, like indole-3-acetic acid (IAA), or 2, 4-dichlorophenoxyacetic acid (2, 4-D) is used as a dedifferentiating hormone for rapid callus induction. Under low auxin concentration and a high cytokinin concentration the cell growth are stimulated and switched concentrations cell division are promoted [10].

4. PROCESS DESIGN CONSIDERATIONS Plant biotechnology requires a interdisciplinary research in plant physiology, cell and molecular biology, pharmacology, toxicology, chemistry, and chemical engineering to assess:

1. Tissue morphology and configuration. 2. Flow and mass transfer environments in

the bioreactor. 3. Kinetics of cell growth and product

formation. 4. Genetic stability of high-producing cell

lines. 5. Control of bioreactor micro and macro

environs. 6. Implications of bioreactor design on

downstream processes. 7. The potential for process scale up.

The goals of plant tissue process development is to achieve high productivity, high product yield and high product concentration by bioreactor operating conditions. Insitu removal of product by extracts of plant cell culture, the use of cell polymer constructs, application of different precursors, elicitors and medium exchange strategies can improve the plant metabolite production [11].

5. BIOREACTORS

A bioreactor is defined as a closed system (vessel/bag or apparatus) in which a biochemical reaction takes place with help of biocatalyst. The biocatalysts is converted into an expressed protein which is biomass or expressed proteins in the process. The term “fermenter” is used only for bioreactors which involves fast growing microorganisms, but in American English this term is used in both bioreactors. The primary role of a bioreactor is to provide containment with optimal conditions for cell growth and formation of products.

5.1 Types of Bioreactors

Suitable bioreactors for plant suspension culture are Stirred bioreactor, Bubble column reactor, Airlift reactor, Bio Wave reactor, Wave & Undertow Bioreactor Slug Bubble Bioreactor, Spray bioreactor, Mist bioreactor. The serious shear-related effects for plant cells generally arise from the aeration and mixing system, the aeration rate or impeller tip speed used. Through sparger rings, air (0.1 and 0.5 v m) is introduced directly, which is located in the lower part of the bioreactor for plant cell cultures. In this way, air bubbles are generated which can damage very sensitive plant suspension cell [12]. It causes bubble bursting. For aerated airlift bioreactors and bubble columns it results in extensive foaming resulting from extracellular polysaccharides, fatty acids and high sugar concentrations in the medium. This can cause the dreaded wall growth phenomenon and blockage of the air exhaust filter, which increases risk of contamination. Ceramic or sintering steel porous spargers, bubble free aeration via tubes of silicone, external aeration via special devices, oxygen supplementation, and changed geometry of the reactor and antifoam agents are suggested to overcome these difficulties [13]. In the case of aeration in high biomass concentrations more than 30 g dry weight, a further difficulty was reported for airlift bioreactors and bubble columns is poor transfer of oxygen and heterogeneous biomass circulation. Therefore


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stirred bioreactors are preferable for cell cultivation, with plants of high densities [14]. An impeller system and its pumping mode for stirred bioreactors is chose on the basis of the aeration system type and arrangement. The location of the sparger in the flow direction of the impeller guarantees mass and temperature homogeneity as well as optimal gas dispersion. Hence, impellers considered have included large slow-moving axial flow impellers with low tip speeds , such as marine impellers or special pitched blade impellers till now, as well as impellers positioned near the vessel wall, for example spiral stirrers, helical-ribbon impellers and anchor impellers. Radial flow Rushton impellers with concave blades are occasionally used. In addition, alternative impeller systems that includes cell-lift impeller, centrifugal-pump impeller have been mounted in stirred vessels [15].

The advantage of low-cost and disposable bioreactors having a gas-permeable cultivation bag of plastic film was effectively proved in a number of plant cell cultivations. Whereas Osmotek’s Life-Reactor and Curtis’ Plastic-lined Bioreactor represent pneumatically driven bubble columns, the BioWave investigated by our team was the first mechanically driven bag bioreactor [16]. Nowadays, further examples of such bag bioreactors is characterized by wave-induced motion (where the mass and energy transfer is manually adjusted via rocking angle, rocking rate, and filling level and culture bag geometry) are available, including, the AppliFlex, Tsunami-Bioreactor, Optima and OrbiCell. A wave bag containing culture medium keep in rocking their platform induces a wave in the bag. While mixing and oxygenation occur, the surface of the medium is continuously renewed and bubble-free surface aeration takes place. The BioWave has a leading position as a result of its scale-up capability to 300 l culture volume and the availability of scale-up criteria based on its hydrodynamic characterization [17]. Moreover, our studies reveal the potential of the BioWave for growing tobacco, grape, apple and yew suspension cells up to 10 l culture volume. We achieved maximum biomass productivities of 40 g fresh weight l-1 d-1 with excellent doubling times of 2 days. Furthermore, savings in time (no preparation, cleaning and sterilization are required), reduced shearing (indicated by higher viabilities, biomass productivities and no significant change in cell morphology), reduced foaming and a lower risk of contamination were found in the Bio-Wave when

compared to cultivations in dominating stirred bioreactor [18]. The recent disposable bioreactor developments, namely the Wave & Undertow Bioreactor and the Slug Bubble Bioreactor, were successfully used to grow tobacco and soya suspension cells expression. There will be shear stress in negligible level [19]. 5.1.1 Disposable bioreactors for In-vitro

culture

For cell culture, the Wave Bioreactor instrument is an effective, cost-efficient device. Culture medium and cells made contact with each other by making contact with a presterile, disposable chamber that is situated on a special rocking platform. Waves in the culture fluid are created by rocking motion of the platform and offers continual mixing and oxygen transfer, resulting in a healthy environment for cell growth. The Bioreactor requires no sterilization, providing comfort of operation and protection against contamination.

5.2 Continuous Stirred Tank Bioreactor A cylindrical vessel with motor driven central shaft that supports one or more agitators were used in continuous stirred tank bioreactors. Usually, agitators also called as impellers. The bottom of the bioreactor contains shaft (Fig. 2A). According to, height to diameter ratio, number of impellers vary. That height to diameter ratio is referred to as aspect ratio. The aspect ratio of a stirred tank bioreactor is generally 3-5. The aspect ratio is less than 2, for animal cell culture. Usually 1/3 rd. of the vessel diameter, gives the diameter of the impellers. The distance between two impellers is approximately 1.2 impeller diameter. Different types of impellers like Rustom disc, concave bladed, marine propeller etc., are in use [20]. In short stirred tank reactors (STRs), the air is added to the culture medium under pressure. A device called sparger is used for aeration. Sparger may contains a ring with many holes or a tube with a single orifice. For better gas division system throughout the vessel, spargers along with impellers are used in this reactors.

Impellers broken down the bubbles generated and discrete throughout the medium. This enables the formation of optimal environment throughout the bioreactor [21].

Fig. 2. A) Continuous stirred bioreactor, B) Bubble column bioreactor, C) Internal loop airlift

bioreactor, D) External loop airlift bioreactor

5.3 Bubble Column Bioreactors

The air or gas is introduced at the base of the column in bubble column reactor, all the way

Su et al.; AJB2T, 5(3): 1-13, 2019; Article no.

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Fig. 1. Wave bioreactor

(C) (D)

Fig. 2. A) Continuous stirred bioreactor, B) Bubble column bioreactor, C) Internal loop airlift bioreactor, D) External loop airlift bioreactor

Bioreactors

The air or gas is introduced at the base of the column in bubble column reactor, all the way

through perforated pipes or plates, or metal micro porous spargers (Fig. 2B). The gas flow rate influences the performance factors like O2 transfer and proper mixing. For better

; Article no.AJB2T.51795

Fig. 2. A) Continuous stirred bioreactor, B) Bubble column bioreactor, C) Internal loop airlift

through perforated pipes or plates, or metal 2B). The gas flow

rate influences the performance factors like ng. For better

performance the bubble column bioreactors may be fitted with perforated plates. In bubble column reactors the vessel is cylindrical. The aspect ratio of this reactor is 4-6 with height to diameter ratio.

5.4 Airlift Bioreactors The medium of the vessel is divided into two interconnected zones by means of a baffle or draft tube in air lift reactors. Among the two zones, one is stated as riser in which the air/gas is pumped. The other zone is the down comer, which receives no gas. The dispersthe riser zone. The down flow occurs in the down comer. There are two types of airlift bioreactors. They are: internal loop airlift reactor and external loop air lift reactor. Internal-loop airlift bioreactor has a single container. Central draft tube that creates interior liquid circulation channels are equipped in that single container. These bioreactors are simple in design, with volume and circulation at a fixed rate for fermentation.

External loop airlift bioreactor (Fig. possesses an external loop. This external loop facilitates the liquid that circulates through separate independent channels. For different fermentation process these reactors are suitablymodified. In general, the airlift bioreactors are more efficient than bubble columns. They are used particularly for denser suspensions of microorganisms. Because in these bioreactors, the mixing of the contents is better compared to bubble columns.

Fig.


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performance the bubble column bioreactors may be fitted with perforated plates. In bubble column reactors the vessel is cylindrical. The aspect ratio

6 with height to diameter ratio.

of the vessel is divided into two interconnected zones by means of a baffle or draft tube in air lift reactors. Among the two zones, one is stated as riser in which the air/gas is pumped. The other zone is the down comer, which receives no gas. The dispersion flows up the riser zone. The down flow occurs in the down

f airlift bioreactors. : internal loop airlift reactor and external

loop airlift bioreactor has a single raft tube that creates interior

liquid circulation channels are equipped in that single container. These bioreactors are simple in design, with volume and circulation at a fixed rate

External loop airlift bioreactor (Fig. 2D) n external loop. This external loop

facilitates the liquid that circulates through separate independent channels. For different fermentation process these reactors are suitably modified. In general, the airlift bioreactors are

olumns. They are used particularly for denser suspensions of microorganisms. Because in these bioreactors, the mixing of the contents is better compared to

For aerobic bioprocessing technology, air lift bioreactors are used. By pumping, thcontrolled liquid flow in a recycle system. Airlift bioreactors are sometimes preferred for methanol production [22], waste water treatment [23], single-cell protein production [24] due to its high efficiency. Airlift bioreactor’s performance idependent on the pumping of air and the liquid circulation [25].

5.4.1 Two-stage airlift bioreactors

For the temperature dependent formation of products, two stage air lift bioreactors are used [26]. Growing cells from one bioremaintained at 30ºC are pumped into another bioreactor [27] which is of temperature 42ºvery difficult to raise the temperature quickly from 30ºC to 42ºC in the same vessel, therefore we use this reactor. Each bioreactors is fitted with valves. The transfer tube and pump are used to connect the bioreactors [28] (Fig. 3). Growing of cells takes place in the first bioreactor and the second reactor is for bioprocessing

5.5 Tower Bioreactors

A tower bioreactor constitutes pressure cycle fermenter with large dimensions. At the bottom of the reactor high hydrostatic stress is created that increases the solubility of O2 in the medium [30]. At the top of the riser, reduces pressure and facilitates expulsion of CO2. The down comer facilitates the back flow of the medium and completes the cycle. The tower bioreactor has high aeration capacities without having moving parts [31].

Fig. 3. Two stage airlift bioreactor


For aerobic bioprocessing technology, air lift bioreactors are used. By pumping, they ensure a controlled liquid flow in a recycle system. Airlift bioreactors are sometimes preferred for methanol production [22], waste water treatment

cell protein production [24] due to its high efficiency. Airlift bioreactor’s performance is

air and the liquid

bioreactors

For the temperature dependent formation of products, two stage air lift bioreactors are used [26]. Growing cells from one bioreactor which is

are pumped into another [27] which is of temperature 42ºC. It is

the temperature quickly from C in the same vessel, therefore we

use this reactor. Each bioreactors is fitted with transfer tube and pump are used to

connect the bioreactors [28] (Fig. 3). Growing of cells takes place in the first bioreactor and the

nd reactor is for bioprocessing [29].

A tower bioreactor constitutes pressure cycle with large dimensions. At the bottom of

the reactor high hydrostatic stress is created that in the medium [30].

At the top of the riser, reduces pressure and . The down comer

flow of the medium and completes the cycle. The tower bioreactor has

es without having moving

5.6 Fluidized Bed Bioreactors

Fluidized bed bioreactor is similar to that of the bubble column bioreactor. But the top position is expanded to reduce the velocity of the fluid in fluidized reactors. The design of the fluidized bioreactors is mainly to facilitate the solids that are retained in the reactor. When solids are retained, the liquid flows out (Fig.4). Fluid suspended biocatalysts such as immobilized enzymes, immobilized cells, and microbial flocs reactions are carried out in these reactors [32].

Fig. 5. Fluidized bed bioreactor


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Fluidized bed bioreactor is similar to that of the bubble column bioreactor. But the top position is expanded to reduce the velocity of the fluid in fluidized reactors. The design of the fluidized bioreactors is mainly to facilitate the solids that

ained in the reactor. When solids are retained, the liquid flows out (Fig.4). Fluid suspended biocatalysts such as immobilized enzymes, immobilized cells, and microbial flocs reactions are carried out in these reactors [32].

Gas is spared to create a suitable gasfluid bed, for efficient function of this reactor. The suspended solid particles are not too light or too dense [33], and they are in a fair suspended state. To maintain continuous contact between the reaction contents [34] and biocatalysts, recycling is needed periodically. It ensures better efficiency of bioprocessing. Fluidized bed reactors also facilitates the efficient transfer of heat to biomass particles [35]. Circulating fluidized beds are employed for biomass fast pyrolysis [36]. However, the scale

Fig. 4. Tower bioreactor

Fig. 5. Fluidized bed bioreactor


red to create a suitable gas-liquid-solid fluid bed, for efficient function of this reactor. The suspended solid particles are not too light or too dense [33], and they are in a fair suspended state. To maintain continuous contact between

ents [34] and biocatalysts, recycling is needed periodically. It ensures better

Fluidized bed also facilitates the efficient transfer of

heat to biomass particles [35]. Circulating fluidized beds are employed for biomass

[36]. However, the scale-up


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Table 1. Types of bioreactors

S. no. Types Driven Aeration Product Starter culture

Advantages Disadvantages Uses

1 Stirred bioreactor

Mechanic

Bubbles/ airlift

- - Easy scale up Simple Well known

Shear stress Mechanic stress Foam

Suspensions

2 Bubble Column Airlift

Pneumatically

Bubbles/ airlift

- - High aeration Low cost High efficient mass transfer

Viscosity Foam Shear stress

Immobilized Suspensions.

3 Hollow fiber pneumatically Perfusion pumps

- - Easy downstream Fast, small Scale, flexible

Contamination. Mass transfer limitation. Low culture of volume.

Adherent cells. Proteins.

4 Orbitally Mechanic Bubbles Airlift absence

18 mg/l Shaken: 20 mg/l

5 L 50 Ml

Simple. Well cost efficiency. Easy to scale up

Only suitable for certain cell lines.

low oxygen cell Lines.

5 Life reactor Pneumatically Bubbles - - Easy scale up Simple. Well known.

Low culture Volume

embriocrops

6 Wave Mechanic bubbles Non detected 5 L for 15% inoculum

Bubble free surface aeration. Well investigated Uniformity of energy negligible foaming.

Rheological Issues of plant culture

Suspension and Immobilized cells

process can be limited by overall heat transfer in this reactor. By adjusting the fluidization gas flow rates, vapor residence time can be controlled; But in bubbling fluidized beds, the undesired biomass entry of [37] particles can occur at high gas velocities. Small, inert sand pprovided in reactor bed to increase the heat transfer rate to the biomass particles. The fluidized bed also contains catalytically active solid particles, which is assimilated in the reactor [38].

5.7 Packed Bed Bioreactors In a packed bed reactors, a bed of solid particles, with biocatalysts on or within the matrix of solids, packed in a column is provided (Figsolids used may be porous or noncompressible or rigid in nature. Over the immobilized biocatalyst, medium is continuously flowed inside the reactors. In packed bed bioreactor, products are released into the fluid and removed. The flow of the fluid can be upward or downward .Mostly down flow under gravity is preferred [39].

The he flow rate of the nutrient broth with increase in concentration of the nutrients. It is rather difficult to control the pH of packed bed bioreactors by the addition of acid or alkali, because of poor mixing. Hence productreactions use these reactors. These bioreactodo not allow accumulation of the prsignificant level [40].

Fig.


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overall heat transfer in this reactor. By adjusting the fluidization gas flow rates, vapor residence time can be controlled; But in bubbling fluidized beds, the undesired biomass entry of [37] particles can occur at high gas velocities. Small, inert sand particles are provided in reactor bed to increase the heat transfer rate to the biomass particles. The fluidized bed also contains catalytically

h is assimilated in the

packed bed reactors, a bed of solid particles, with biocatalysts on or within the matrix of solids, packed in a column is provided (Fig. 6). The solids used may be porous or non-porous gels, compressible or rigid in nature. Over the

, medium is continuously flowed inside the reactors. In packed bed bioreactor, products are released into the fluid and removed. The flow of the fluid can be upward or downward .Mostly down flow under

rient broth with increase in concentration of the nutrients. It is rather difficult to control the pH of packed bed bioreactors by the addition of acid or alkali, because of poor mixing. Hence product-inhibited reactions use these reactors. These bioreactors do not allow accumulation of the products to any

5.8 Photo-bioreactors

Photo bioreactors are closed systems. Photo bioreactor working either outdoors or indoors.photo bioreactors, single species is inoculated to keep a clean-culture operation. Closed cultivation systems prevents contamination. It leads to higher growth and quality of the product harvested [41]. Manufacturing costs increases for photo bioreactor’s products. Some process can be carried out either by exposing to sunlight or artificial illumination. Since artificial illumination is expensive, therefore outdoor photo-bioreactors are preferred. Important compounds are produced by using photobioreactors are: p-carotene, asthaxanthin. The different types of photo-bioreactors are depicted in Fig. 7. Glass or more commonly transparent plastic are used for making this reactors. The arrangement of tubes or flat panels establish light receiving systems, which is commonly known as solar receivers. Through the solar receivers culture can be circulated by methods such as using centrifugal pumps or airlift pumps. It is necessary to keep cells in continuous circulation without forming any sediments. Adequate penetration of sunlight should be maintained continuously. To prevent rise in temperature, the tubes should be cooled periodically.bioreactors are usually operated in a continuous mode and temperature should be maintainedthe range of 25-40ºC. Microalgae and cyanobacteria are normally used to cultivate in photo bioreactor. The products are produced in night time and the cell grows in daylight [42].

Fig. 6. Packed bed bioreactor


are closed systems. Photo bioreactor working either outdoors or indoors. In photo bioreactors, single species is inoculated to

culture operation. Closed cultivation systems prevents contamination. It leads to

uality of the product Manufacturing costs increases for

photo bioreactor’s products. Some fermentation process can be carried out either by exposing to sunlight or artificial illumination. Since artificial illumination is expensive, therefore outdoor

erred. Important compounds are produced by using photo-

carotene, asthaxanthin. The bioreactors are depicted

in Fig. 7. Glass or more commonly transparent plastic are used for making this reactors. The

of tubes or flat panels establish light receiving systems, which is commonly known as solar receivers. Through the solar receivers culture can be circulated by methods such as using centrifugal pumps or airlift pumps. It is

inuous circulation without forming any sediments. Adequate penetration of sunlight should be maintained

To prevent rise in temperature, the tubes should be cooled periodically. Photo-bioreactors are usually operated in a continuous

temperature should be maintained in C. Microalgae and

cyanobacteria are normally used to cultivate in photo bioreactor. The products are produced in night time and the cell grows in daylight [42].

Fig. 7. Photo bioreactor

Fig. 8. Mist reactor

5.9 Mist Reactor

The cultivation of hairy roots with high potential is done by the gas phase reactors called mist reactors. The roots have an optimal growth, critical droplet size is between 1µm and 35 µm Liquid nutrients are homogeneously distributed in this case and the gas transfer into the rots is does not experience oxygen stress.vessel with stainless steel upper and lower lids is equipped in mist reactors. The vessel stands vertically on four legs. In the bioreactor’s lower lid thermos regulation system is located. Sensors for temperature and levelling are locatebioreactor’s upper lid [43]. It is supplied with a LED jacket, a hydraulic spray nozzle, three sample holder plates and a ring sparger. Gas inlet/outlet ports are located on the upper lid of the bioreactor. The exhaust gas condenser is fixed on the upper part of the bioreactor. It prevent unwanted liquid particle flowing out frothe vessels environment. For easier autoclaving the design of the condenser provides an option


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Fig. 7. Photo bioreactor

The cultivation of hairy roots with high potential is done by the gas phase reactors called mist reactors. The roots have an optimal growth, if critical droplet size is between 1µm and 35 µm Liquid nutrients are homogeneously distributed in this case and the gas transfer into the rots is does not experience oxygen stress. A glass vessel with stainless steel upper and lower lids is

ist reactors. The vessel stands vertically on four legs. In the bioreactor’s lower lid thermos regulation system is located. Sensors for temperature and levelling are located on the

It is supplied with a spray nozzle, three

sample holder plates and a ring sparger. Gas inlet/outlet ports are located on the upper lid of the bioreactor. The exhaust gas condenser is fixed on the upper part of the bioreactor. It prevent unwanted liquid particle flowing out from the vessels environment. For easier autoclaving the design of the condenser provides an option

for the user to adjust its position. The condenser is provided with an adjustable gas valve for pressure regulation [44].

5.10 Low Cost Mist Bioreactor (LCMB) The largest gas-phase bioreactor with a capacity of 60 L. Low cost mist bioreactors were designed to grow Artemisia annul transformed roots and Dianthus caryophyllus shoots. The reactors use similar mist generators but the culture chambers were modified to meet the requirements of each application [45].

5.11 Wilson Bioreactor The largest hybrid bioreactor system with a capacity of 500 L. The spray reactor reminds a gas-phase bioreactor with a cultivation container with horizontal meshes. This bioreasome problems such as very laborious handling and often associated with contamination issues, now is no longer in use [46]. 5.12 Balloon-type It is a spherical-glass bioreactor, which is used for root culture due their advantages of easy moisture and simplicity, although these assets are no-longer in use due to easy contamination and difficulty of take out the roots useful for extracellular products [47].

6. CONCLUSION A variety of bioreactor types providing growth and expression of bioactive substances are developed for plant suspension culturePlant cell suspension culture as a bio production platform provides a number of unique advantages for recombinant protein production [49]. These bioreactors types used for synthesisof complex chemicals whereas low biomass and product level from plants can be achieved.

In the early past, industries use stirred bioreactors, hollow fibers and bubble columns for their production as it’s possible to see in Phyton Biotech [50], which uses bioreactors of stainless steel. But now, single-use bioreactors are gone to set aside this last device, Phytocelltec [51] uses Malus Domestica for obtaining a component for a liposomal active ingredient based on stem cells from the Tutwiler Spat Lauper apple, this cosmetic industry uses bio wave bioreactors in sequence instead of huge stainless-steel bioreactors [52]. Single


for the user to adjust its position. The condenser is provided with an adjustable gas valve for

5.10 Low Cost Mist Bioreactor (LCMB)

phase bioreactor with a capacity of 60 L. Low cost mist bioreactors were designed to grow Artemisia annul transformed roots and Dianthus caryophyllus shoots. The reactors use similar mist generators but the culture chambers

d to meet the requirements of each

The largest hybrid bioreactor system with a capacity of 500 L. The spray reactor reminds a

phase bioreactor with a cultivation container with horizontal meshes. This bioreactor has some problems such as very laborious handling and often associated with contamination issues,

glass bioreactor, which is used for root culture due their advantages of easy moisture and simplicity, although these assets

longer in use due to easy contamination and difficulty of take out the roots useful for

A variety of bioreactor types providing growth bioactive substances are

developed for plant suspension culture [48]. Plant cell suspension culture as a bio production platform provides a number of unique advantages for recombinant protein production [49]. These bioreactors types used for synthesis of complex chemicals whereas low biomass and product level from plants can be achieved.

In the early past, industries use stirred bioreactors, hollow fibers and bubble columns for their production as it’s possible to see in Phyton

es bioreactors of use bioreactors

are gone to set aside this last device, Phytocelltec [51] uses Malus Domestica for obtaining a component for a liposomal active ingredient based on stem cells from the Tutwiler

er apple, this cosmetic industry uses bio wave bioreactors in sequence instead of

steel bioreactors [52]. Single-use


11

bioreactors are a reality in R+D laboratories due to their high amount of data in less time, low cost and safety but in industry stainless steel bioreactors are yet implemented; however low-cost safety, short time of production (it’s not necessary to get clean and disinfected), high flexibility and simplicity are going to change this trend. Products as their culture types are different, consequently the disposable bioreactors are lots [53], it seems that stirred bioreactors with different impellers are the most used kind of device, it has plenty of advantages and low cons, so the combination of stirred devices with bubble columns or Airlifts, suggests a great solution [54]. Orbital bioreactors have more advantages than stirred but they only can be used for low-oxygen cell lines. Hydraulic and hollow fibers are only focused in Immobilized and attached cells. Root culture, due their difficulty and own characteristics has specific bioreactors, now the most used is the Mist bioreactor [55], Wilson bioreactor have a great capacity and product titer but it has plenty of contamination. In the other hand, single use bioreactors, stirred ones have importance, but their lack in no foam formation and shear stress improves the acceptance of wave-mixing bioreactors, the only problem of this kind of device is the rheological issues of plant cultures, but this could be avoided by rise the rocking angle and velocity.

COMPETING INTERESTS Authors have declared that no competing interests exist.

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