CA2801768A1

CA2801768A1 - A photobioreactor bag with built-in sparger tube, agitator and water

Info

Publication number: CA2801768A1
Application number: CA 2801768
Authority: CA
Inventors: Soheyl S. M. Mottahedeh
Original assignee: Individual
Current assignee: Individual
Priority date: 2013-01-09
Filing date: 2013-01-09
Publication date: 2014-07-09

Abstract

The invention features a pond-like photobioreactor that combines a reusable bioreactor bag with a translucent elongate semi-rigid container that lines it. Agitation is provided, in one embodiment of the invention by alternating gas flow between two built-in sparger tubes formed from the bag itself.
In another embodiment, two elongate side pockets bordering the bioreactor bag provide the double function as water jacket for controlling temperature and as wave generator when inflating them alternatingly. In addition to aeration, agitation, cultivation and temperature control the bag isolates a portion of the medium for harvesting secretions of cyanobacteria when the bag is combined with a container having multiple recesses, one of them containing an inflatable system.

Description

A PHOTOBIOREACTOR BAG WITH BUILT-IN SPARGER TUBE, AGITATOR
AND WATER JACKET
FIELD OF THE INVENTION
This invention relates to the field of photobioreactor bags for culturing algae and cyanobacteria. In particular, this invention combines a pond-like elongate container enclosing a reusable photobioreactor sleeve that incorporates two sparger tubes, an agitation system and a water jacket for temperature control, all built-in into the bag.
BACKGROUND OF THE INVENTION
Current photobioreactor equipment are often complex and expensive. These factors make their use valid only for production of expensive pharmaceuticals, nutraceuticals and chemicals.
To address environmental or energy issues such as need for capturing of carbon dioxide emissions, treatment of waste waters, food security, shortness of fresh water, need for renewable energy, cost of photobioreactors must come down For this, major technology breakthrough are needed.
Furthermore, for large-scale culture of algae and cyanobacteria to become viable, agricultural farmers experienced with sustainable large-scale farming may need to become involved in algae farming.
Supplementing their crops with algae farming will make the process even more sustainable. For this, the complexity of algae farming must be reduced to a minimum, scale-up has to become within reach and costs have to be reduced by many folds.
The present invention therefore represents a fresh solution to the aforementioned problems, providing a low-cost photobioreactor system that integrates the scalability of ponds with the controls provided by photobioreactors. In the present cultivation system, a low-cost semi-disposable bioreactor bag is provided with all the controls of photobioreactors and is made to line the interior space of a durable pond-like container, thus enabling the large-scale production of algae and cyanobacteria in a controlled environment by people not specifically trained in microbiology or aseptic technique.
The reusable, modular bag of the present invention, while essentially disposable, may be used continuously for multiple consecutive culturing/harvesting cycles.
The multiple features of the modular bioreactor bag of the present invention when associated with the versatility in shapes provided by an associated container that encloses the bag, enables the bag to aerate, agitate, cool, cultivate, milk cyanobacteria and harvest secretions in an all-in-one bioreactor bag, thus keeping sterility and costs of the process within control.
When eventually the reusable bioreactor bag does become contaminated, it may then be disposed of with relatively little economic loss. Such modules may be cheaply manufactured, even for production volumes of 1,500 liters or more of culture. Further, the ability to perform a number of culturing/harvesting cycles is economically lucrative, lowering even further the effective cost per module. A farm of such modular bags in associated containers can be economically arranged, and the number of modules in the farm may be controlled to closely match production to demand. Thus, the transition from pilot plant bioreactors to large scale production may be achieved in a relatively simple and economic manner by adding more modules to the farm.
Furthermore, eliminating the need for external sparger piping systems, for external motive device to agitate the medium, for external heat exchangers to control temperature, for pumps to flow a portion of the medium to milk cyanobacteria and aseptic equipment to harvest the rich secretions, enables to keep capital costs down and attracts more investors and farmers to invest in algae farming.
SUMMARY OF THE INVENTION
According to a preferred embodiment of a combination bag/container photobioreactor for culturing and agitating algae and cyanobacteria in a medium, the bag comprises a flexible light transmitting plastic sheet forming a closed chamber that further includes at least two alternately pressurized sparger tubes spaced apart from each other and formed from the light transmitting plastic sheet itself. The two sparger tubes are located preferably at locations not difficult to effect circulation of medium with bubbles alone.
For generally oval-shape containers, sparger tubes are located on the bioreactor close to bag extreme borders. For bioreactor bag associated with container having multiple recesses, the distance between sparger tubes may vary according on the size and shape of the bioreactor, but preferably, tubes may be located as far as possible to each other as long as they don't interfere with the bioreactor's operation such as culturing, milking and harvesting in the case of cyanobacteria.
While one end of sparger tubes is closed, the other end is in fluid communication with a three-way valve that alternately diverts incoming gases to each tube in a timely manner.
This alternating flow creates pressure variation from two opposite directions and causes increased agitation of the medium.
To further increase agitation's amplitude, an elongate pivotable flap comprised of a strip of flexible plastic that extends along the bag length is bonded on one edge end to the plastic sheet with the other edge end being positioned above a sparger tube; as a sparger tube is inflated and deflates by incoming flowing gases, the flap moves cyclically up and down increasing agitations of the medium.
By coordinating movement of two flaps in a timely manner, agitation is further amplified.
According to some embodiments of the bioreactor, the bag includes two side water jackets for circulating a conditioning fluid to control temperature of the medium in the bag. Inflating alternatingly these two side water jackets generates waves, solar reflector film bonded to the bottom portion of the bag added side layer increases sunlight penetration from the bag two sides.
According to some preferred embodiments, the bag cooperates liningly with the inner surface of a semi-rigid flexible translucent W-shape container configured with multiple recesses. This arrangement associated with an expandable chamber in one of the recesses, enables to raise or lower the level of the medium in the bag. Deflating the expandable chamber in a main recess causes the medium to retrieve in that main recess while portions of the medium sunk in other recesses becomes isolated for being processed, for example for milking cyanobacteria.
The present invention also discloses a method of building a sparger tube into a closed bag. This method teaches to first perforate near one edge a folded portion of a bag laid in a layflat position, then tuck in this perforated portion to form a gusset and finally heat bond the newly created gusset edge to seal the bag.
Thus, a bioreactor bag of the present invention may incorporate concurrently a sparger tube, an agitation system, a temperature control system, a medium isolation system, a bacteria milking space, and a harvesting, all-in-one built in the same bag, eliminating the need for costly accessories.
Other features and advantages of the present invention will be better understood by reference to the drawings and detailed description that follow.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 Perspective view of a photobioreactor bag with two built-in sparger tubes, delivering gases alternately.
FIG. 2 Close-up view A of the cross-sectional view of sparger tube of FIG. 1 illustrating how gases exit sideways.
FIG. 3 Cross-sectional view of a photobioreactor bag and two side water jackets inside a container with two sparger tubes each enclosing a flap.
FIG. 4 Cross-sectional view of a photobioreactor bag and two side water jackets inside a container with two sparger tubes each activating an external flap welded to the bag.
FIG. 5 Cross-sectional view of a photobioreactor bag and two side water jackets inside a container with two sparger tubes each activating an external flap enclosed in a closed tube.
FIG. 6. Cross-sectional view of a photobioreactor bag lining a reverse U-shape container enclosing an expanded height-adjustable chamber.
FIG. 7. Cross-sectional view of photobioreactor bag of FIG. 6 with a retracted chamber.
FIG. 8. Perspective view of compacting roller over a land shaped for supporting a bioreactor container.
FIG. 9 Perspective view of compacting roller over a land shaped for supporting a W-shape bioreactor container FIG. 10 Perspective view of a W-shape bioreactor container resting over a shaped land.
FIG. 11 Perspective view of a method for building a sparger tube in a bioreactor bag.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a combination photobioreactor bag lining a trough-like container for culturing microalgae cells and cyanobacteria organisms. The combination is easy to use, inexpensive and versatile. The invention enables the preparation of microorganisms by people such as rural and urban algae farmers not specifically trained in microbiology or aseptic technique. It enables microorganisms to be grown safely, for a variety of purposes, without the need for specialized facilities such as temperature controlled rooms, externally driven agitators, external insertable aeration and cooling pipes and other associated accessories.
The photobioreactor bag of the present invention, also referred interchangeably as bioreactor, bag chamber and sleeve are designed for culturing a wide variety of microorganisms, both aerobic and anaerobic. They are suitable for culturing microorganisms in generally sunny and environmentally friendly temperatures, and preferably in arid lands and other locations not competing with food production. They can also be used for fermentation of organic and non-organic products including fruits, vegetables and grains; or be used for growing marine organisms as small as 1 micron to as big as larvae and smaller fishes. Some of microorganisms that may be successfully cultured in the present reusable bioreactor bags of the present invention include, but not limited to, bacteria, cyanobacteria, fungi, algae, protozoans and nematodes.
Referring now to the drawings, FIG. 1 shows a preferred embodiment of the photobioreactor bag 10 made of a light transmitting flexible plastic sheet 12 forming an elongate closed chamber lining a generally horizontal translucent surface provided with optional recesses, including two sparger tubes 22 and 24 which are formed by shaping portions of plastic material 12.
FIG. 2 illustrates a close-up view of the sparger tube 22 perforated with two side holes for exiting gases such as air and carbon dioxide. The bioreactor bag 10 is provided with multiple ports (not shown) for introduction or removal of gases or liquids. These ports are located at the transversal ends of plastic sheet 12.
FIG. 3 shows in a preferred embodiment of a combination bioreactor bag 10 lining the interior of a translucent C-shape container 40. In this illustration, agitation is provided with flaps 16 and 18 being encased in sparger tube 26 and 28 having holes on one side of the tube. Flaps 16 and 18 increase amplitude of the agitation provided by bubbles exiting from sparger tubes 26 and 28.
FIG. 3 also illustrates two side water jackets 32 and 34 that envelop from one side the shape of the centrally-located bioreactor bag 10 and from the other side adopt the interior shape of the translucent container 40 that supports and contains all three bags 32,10, and 34. Water jackets 32 and 34 contain fluids that are displaced therein for controlling temperature of medium 50 contained in bioreactor bag 10. Inflating alternatingly water jackets 32 and 34 generates waves.
Additionally, fluids in water jackets 32 and 34 may contain electrochromic polymers or nanoparticles 62 that electrically change their color to reduce bag 10 exposure to excess sunlight or to provide an on-off filtering effect, such as biotuning by flickering the color change at desired frequencies to enhance algae and cyanobacteria growth. Water jackets 32 and 34 may be made of the same or of a different material than the light transmitting plastic sheet used for bioreactor bag 12.
The translucent container 40 illustrated in FIG. 3 is made of a semi-rigid flexible plastic such as, but not limited to, a fiberglass sheet 40 provided with a plastic memory C-shape designed to keep its trough-like borders in an elevated position, like a container. Wedges 70 further assist container 40 to maintain and reinforce its trough-like shape. Container 40 is made of a semi-rigid material such as fiberglass or of thin plastic sheet that is bendable so that the container 40 elevated borders may flatten when container 40 is rolled-up, thus reducing costs in packaging, transport, installation and maintenance.
Furthermore, FIG. 3 illustrates a solar reflector 60 positioned underneath the translucent container 40.
This reflector 60 directs added solar reflection to container 40 two sides and partially to the container 40 underside. To prevent container 40 from rocking sideways bricks or wedges 70 block movement of container 40.
FIG. 4 shows a preferred embodiments of a combination container and bioreactor bag 10 wherein spargers tubes 26 or 28 expand when pressured by flowing gases. This causes to lift pivotable flaps 16 or 18. The welding of flap 16 to plastic sheet 12 along the flap 16 one elongate edge creates a spring effect bringing back flap 16 to its lowest position. By alternating gas flow from one sparger tube 26 to another sparger tube 28, flap 16 and 18 are radially moved, causing agitation of medium 50.
Using a programmable timer to alternate an incoming gas flow in flow communication with sparger tubes 26 and 28 via a three-way valve (not shown) enables to harmonize the interplay between flaps 16 and 18 in order to maximize agitation amplitude which leads to the creation of waves. This flow alternation between two sparger tubes 26 and 28 eliminates the need for an external drive that agitate a medium as practiced in prior art.
FIG. 5 shows in an embodiment of a combination container 40 and bioreactor bag 10, wherein a gusseted sealed tube 20 encloses a flap 16 that is resting over a sparger tube 26. Inflating sparger tube 26 causes radial pivoting and lifting of flap 16. This in turn causes agitation of medium 50.
Keeping the sealing line portion of the sparger tube 26 as close as possible to the bonded edge of the gusseted tube 20 increases the amplitude of flap 16 by virtue of the leveraging effect. By alternating gas flow from one sparger tube 26 to another sparger tube 28, flap 16 and flap 18 (not shown) respectively pivot, causing increased agitation of medium 50. Using a programmable timer to control gas flow via a three-way valve (not shown) enables to harmonize the interplay between flaps 16 and 18 in order to maximize amplitude of waves. This arrangement eliminates the need of external drive that agitate a medium as practiced in prior art.
FIGS. 6 and 7 show some embodiments of the combination container 40 and bioreactor bag 10 wherein the level medium 50 is made to vary as per need. This embodiment of the invention may be preferably used for culturing, milking cyanobacteria and harvesting secretions in a single bioreactor bag 10. In this embodiment, bioreactor bag 10 further includes an expandable chamber 36 cooperating with a translucent container 40 configured with a generally flat surface 80 flanked between two recesses 82 and 84. Chamber 36 may be created as an integral part of plastic sheet 12 formed by an elongate sealing line that separates bag 12 major portion from a chamber portion 36.
This chamber 36 may optionally be positioned inside bag 12 resulting from the creation of an internal sealed gusset. Alternatively, this chamber 36 may be annexed to bag 12 as a separate tubular, flexible expandable plastic container 36 in wall contact with bag 10. In this embodiment, height of medium 50 in bag 12 can be elevated above the level of flat surface 80 when chamber 36 is fully expanded maximizing surface exposure of medium 50 to sunlight.
Similarly, as shown in FIG. 7, level in medium 50 can be lowered below flat surface 80. To achieve this, chamber 36 in emptied from its fluid content causing in turn medium 50 to retrieve itself driven by gravity force into the bag portion present in recess 82. This causes in turn one or multiple portions of medium 50 trapped into one or more other recesses, such as in recess 84, to become isolated from the medium portion retrieved in recess 82. Once isolated, medium portions in each recess may be processed separately.
As an example of such a process, the bag portion associated with recess 84 may contain highly porous matrices or gel, such as gel silica 90, for entrapping cyanobacteria.
Subjecting said entrapped cyanobacteria to conditions that trigger secretions, such as, but not limited to an acidic condition, through injection of a product via one or multiple entry ports associated to said recess portion 84 causes cyanobacteria to release secretions that can be collected and retrieved through associated exit ports.
FIG. 7 shows other processes associated with a same or a separate recess 82 and 84. These processes may include, but are not limited to, exposing medium 50 to intense light of selected wavelength 92, to heat, to cold, to electromagnetic fields 94, to osmosis exchange, to catalysts, to cell density counting, to physical measurement, to continuous removal of secondary metabolites, to bio organic solvents, to electrocatalysts, to biocatalytic reductions, to fermentation and to a combination thereof.
The bag portion present in recess 82 containing height-adjustable chamber 36 may be insulated from outside environmental conditions such as, but not limited to, excess heat, excess cold, excess light, excess wind, or a combination thereof.
Medium 50 contained in the bag portion associated with some of the recesses such as 82 and 84 may be further subject to downstream processes such as, but not limited to, dewatering, harvesting, extraction, bacteria milking, collection of secretion or to a combination thereof.
FIGS. 8 and 9 show an embodiment of an embossed surface acting as support for container 40. The configuration of such an embossed support enables light penetration to support 40 from all directions.
To emboss a C-shape, a W-shape or other shapes in land 200, a soil compacting roller 100 is provided with protrusions and recesses. These are configured to provide protrusions that will support intermittently the bottom of the translucent container 40 for containing the heavy medium content 50.
configuration having and recesses that enable light entry from all directions around container 40. It is known that dirt roads in rural areas are stabilized by various means such as by addition of lime, cement or enzymes to withstand harsh environmental conditions. Similarly, supports made of embossed dirt and clay may be stabilized to withstand environmental conditions. To bring added light to underside of container 40, recesses may be covered with a solar reflector material such as a solar reflector film 60 or a reflective mineral.
FIG. 10 shows a land surface 200 embossed with protrusions and recesses configured for supporting a W-shape container 40. Such a configuration is particularly suitable for growing cyanobacteria as the container surface is provided with recesses that enable to isolate medium 50 for milking bacteria using a single bioreactor bag 10 rather than requiring multiple accessories and equipment as practiced in practiced in prior art.
FIG. 11 shows an embodiment of plastic sleeve 2 being processed by various equipment for building a sparger tube 22 in sleeve 12. The continuous manufacturing process can be applied in-line with a blown film extrusion process or applied post extrusion after the film is made.
In both cases, the folded plastic film or sleeve 12 is first drawn over a perforating equipment 120, such as a punch or a needle wheel, then is pushed by a gusseting wheel 130 to tuck the perforated portion inside bag 12 before a sealing machine 140 bonds the newly formed edge. Sealing may be performed using ultrasonic, heat or radio-wave welding 140.
The process of FIG. 11 creates a sparger tube 22 with two side-holes. This overcomes disadvantages present in prior art with single-hole sparger tubes that shoot gases away from a bioreactor surface and creates "dead space" around single-hole sparger tubes and where it would be difficult to effect circulation with bubbles alone. In the present invention, sparger tubes 22, 24, 26 and 28 exit bubbles sideways, slightly downwards, just slightly above tube's sealing line. The generally oval-shape of bioreactor bag 10 eliminates corners, therefore minimizing settling of microorganisms. Similarly, sparger tubes such as 26 and 28 located on recess walls of W-shape bioreactors shoot in both directions therefore dislodging cells in dead spaces.
In a standard photobioreactor, two means are used to achieve proper aeration and optimum exposure of micro organisms to light (1) bubbling of gases through the growth medium and (2) agitation by a pump, a stirrer or other means to effect a mechanical circulation. Using the present photobioreactor 10 , aeration and mechanical agitation are combined into a single process ¨ this maximizes the use of existing components for creating a multifunctional bioreactor 10, thus avoiding the need of a pump or a stirrer that add capital, maintenance and operation costs with additional use of energy. This has been accomplished by coordinating alternately generation of gas bubbles from two opposite directions exiting from two sparger tubes spaced apart. In this bioreactor 10, each sparger tubes 22 and 24 also project gas bubbles from two opposite directions. Thus, in this bioreactor 10, four sources of bubbles provide aeration, mixing and agitation without need of additional mechanical equipment, air supply or energy except for operating a small controller that operates a three-way valve.
The bioreactor bag 10 is constructed from flexible or semi-flexible, waterproof material, preferably plastic. Low Density Polyethylene LDPE has been used in the instant bioreactor, but other kinds of plastic such as, but not limited to, High Density Polyethylene (HDPE) are suitable, and may be desirable in special circumstances. Plastic may be chosen for light transmitting qualities. Certain cells may be sensitive to bright white light or ultraviolet radiation, and for these cells the container can be made of plastics that absorb these wavelengths. Likewise, other kinds of cells, plant cells for example, may be developmentally regulated by the spectral quality of light, and for these cells the bioreactor bag 10 can be made of plastics that selectively transmit the desired wavelengths of light.
The strength of the plastic is also an important consideration. Different thicknesses of plastic can be used, according to end purposes and standard practice. For example, for single-use bioreactor bags thicknesses between 75 microns to 150 microns may be sufficient. For multiple-use bioreactor bags 10, thicknesses may vary from 100 micron to 300 microns.
Shaping thin, durable fiberglass container 40 with a plastic memory shape to contain a bioreactor bag 10 is not only cost-effective but protects the bag from harsh environmental conditions by providing it with very durable skin. Manufacturing complex-shape containers such as a W-shape container 40 with multiple recesses may be accomplished by joining transversally, side-by-side shaped fiberglass sheets that have one recess in each sheet. This enables also to increase the width of the bioreactor 10.
Accordingly the overall size of the bioreactor bag width shall be adjusted when using a wider container.
Inserting two pre-shaped fiberglass sheets with plastic memory shape into each other creates a container 40 that has reinforced structure. Inserting a spacer for creating a space between the two pre-shaped fiberglass sheets enables to create a water jacket for circulating a temperature conditioning fluid in between them. Inserting a reversed pre-shaped sheet that has plastic memory to envelop the borders of another pre-shaped sheet creates a closed container 40 with excellent resistance to harsh weather conditions.
The container 40 includes one or more ports to serve as inlets or outlets for gases or liquids. The ports are constructed of rigid or semi-rigid materials that are compatible with the material used for construction of bag 10. In preferred embodiments, any standard plastic tubing or molded plastic can be used to construct the ports, and they are welded into the seams of the container according to standard techniques. A variety of such ports are commercially available, e.g., for use in medical bags and similar water containers.

The disposable bioreactors of the present invention preferably are pre-sterilized prior to shipping to the local user. Various sterilization techniques may be used. Sterilization techniques that are common in the art include, but are not limited to, dry heat, autoclaving, radiation, and ethylene oxide gas.
Air or other gases are introduced into the bioreactor bag10 via a gas inlet port. Tubing connecting the port to a gas pump is fitted with at least one filter, for filtering microbes from the airstream prior to introduction of the gas into the bioreactor bag 10.
The bioreactor bag 10 comprises a vent to exhaust gases. The vent in the bioreactor bag 10 of the present invention preferably comprises an outlet port fitted with tubing, of varying length.
For convenience and ease of use, the reusable bioreactor bag 10 of the present invention is packaged and sold as a kit. Typically, a kit can contain one or more reusable bioreactor bags (e.g., 1500 L in size), a 15m long and lm wide container with a C-shape configuration by virtue of its plastic memory shape imparted during manufacturing. When rolled, the elevated walls of the C-shape container flatten and regain their original flat dimension of1.2m wide. Also is included in the kit, a programmable three-way valve for alternating flow of incoming gases between the two sparger tubes in bioreactor bag 10. An optional air pump is also included. The user supplies water, electricity, wedges such as bricks to keep the container borders elevated and a land space to support the combination container / bioreactor bag. Not only the length of the container can be increased but also the width of the container can be augmented by joining multiple sheets side-by-side. Accordingly, the size of the bag can also be increased to line the associated container.
To operate the combination container / bioreactor bag, the user unrolls the container which automatically takes its plastic memory shape and places the elongate bag or sleeve inside the container by also unrolling it. If temperature control is required, a separate plastic bag with two side water jackets or the side pockets in the bioreactor bag are filled with water at a desired temperature.
The filling is done by connecting a water hose to the jacket entry port. The jacket exit port is either connected to an evaporative cooler or to other temperature conditioning device, which in turn connects to the jacket entry port.
Then, the user connects a first gas inlet port with tubing containing an in-line micro-filter to sterilize the air to one side to the air pump and the other side via an air flow meter to a first entry port of a double entry connector; the hose carrying CO₂ is connected via a CO₂ flowmeter to the second entry port of the double entry connector. The double entry connector feeds gases to entry of the the three-way valve. The user then connects the two exiting hoses from the three-way valve to the two sparger tubes entry ports attached to the bag. The user fills the bioreactor bag with a medium comprising of tapwater (or sterilized sea water) mixed with nutrients, via faucet or garden hose using the existing third inlet port. The incoming water is sterilized via one or more in-line filters (e.g., 5 µM
pre-filter obtained from a household water filtration unit, combined with a microbiological final filter (0.45 µM). The exhaust port, namely for 0₂ is fitted with an exhaust tube. Finally algae or cyanobacteria inoculum is introduced in the bag before electricity is turned on.
Once the combination container / bioreactor bag kit is filled with medium, inoculated and connected to the air pump, the microorganisms are cultured for a pre-determined amount of time, under specified culture conditions, according to standard procedures. When the culture is complete the contents are harvested, e.g., by draining the container through one of the bag exit ports.
The aforementioned bioreactor kit has been exemplified for the cultivation of algae and cyanobacteria.
However modest adjustments can adapt the kit for use with many other cells and organisms.
Adjustments include different media and inoculum formulations, all of which would be known to one skilled in the art.
The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The described implementations are to be considered in all respects only as illustrative and not restrictive. The scope of the invention is, therefore, indicated by the appended claims rather than by the foregoing description. All changes which come within the meaning and range of equivalency of the claims are to be embraced within their scope.

Claims

1. A photobioreactor combining a reusable bioreactor bag lining a durable container, said combination comprising:
a generally horizontal, translucent, wide, elongate, flexible, semi-rigid container;
a bag lining said container for culturing and agitating an algal medium therein; said bag comprising a self-standing flexible light transmitting plastic sleeve forming an elongate closed chamber;
the plastic sleeve further including at least two sparger tubes positioned at locations favorable to effect circulation of medium with bubbles alone, such as on the chamber floor or near walls;
said sleeve one end is provided with at least three inlet ports comprising one in fluid communication with said chamber and two inlets in fluid communication with said two sparger tubes, and other end with at least two outlet ports comprising one in fluid communication with the chamber and one for removal of gases; and a source of gases in alternating flow communication with each of the sparger tubes, said flow alternation between the two tubes amplifying agitation in the medium.

2. The combination of claim 1, wherein said sparger tubes are continuously formed from said plastic sheet; each of said tubes created by first perforating an elongate optionally one-layer or folded sheet and then tucking said perforated portion for creating pleats which edges are bonded together to form a perforated closed tube inside said self-standing plastic sleeve.

3. The combination of claim 1, wherein the medium consists of bacteria, cyanobacteria, fungi, algae, protozoans, nematodes or a combination thereof.

4. The combination of claim 1, wherein said container is made from, but not limited to, semi-rigid flexible sheets, such as, but not limited to, a fiberglass sheet of about 0.6m to about 2.5m wide, about 0.5mm to about 1.3mm thick.

5. The combination of claim 1, wherein said container shape includes multi-radius cross-sectional shapes such as, but not limited to, a C-shape, W-shape or other shapes including multiple recesses; said shapes optionally fixed imparted during manufacturing to become plastic memory or gained by bending and enclosing said sheet into multiple adjacent multi-radius shape brackets.

6. The combination of claim 1, wherein two of said pre-shaped sheets having plastic memory reinforce each other when one is inserted into the other.

7. The combination of claim 5, wherein said two inserted plastic memory further include a spacer for creating a space therebetween, said space for circulating a temperature conditioning fluid therein.

8. The combination of claim 4, wherein two of said pre-shaped sheets having plastic memory form a closed container when one pre-shaped sheet is reversely enveloping the borders of the other.

9. The combination of claim 1, wherein each of said sparger tubes one end is sealed and the other end is in fluid communication with a controllable three-way valve alternatingly diverting gases to each tube.

10. The combination of claim 1, further including at least two pivotable flaps, each pressingly contacting on one elongate border one of said sparging tubes; said flap comprising an elongate semi-rigid strip of plastic extending along the sparger tube length and being bonded on the other elongate border to said light transmitting plastic sheet; each of the flaps being moved radially by contacting sparger tube that expands and contracts cyclically; thereby amplifying medium agitation as bubbles exit.

11. The combination of claim 1, further including at least two flaps, each enclosed in a sealed gusseted tube, each gusseted tube resting over one of said sparging tubes;
each of said flaps comprising an elongate semi-rigid strip of plastic extending along the sparger tube length and being enclosed in said sealed gusseted tube formed from said light transmitting plastic sheet;
each flap being moved radially by underneath sparger tube that expands and contracts cyclically; thereby amplifying medium agitation as bubbles exit.

12. The combination of claim 1, further including at least two flaps, each enclosed in a sparger tube formed from said light transmitting plastic sheet, each of said flaps comprising an elongate semi-rigid strip of plastic extending along the sparger tube length;
each flap moving radially as sparger tube one side bonded to said chamber wall expands and contracts cyclically; thereby amplifying medium agitation as bubbles exit.

13. The combination of claims 1 to 5, further including at least two elongate light transmitting sleeves containing a flowing temperature-controlling fluid therein forming two water jackets, each positioned on one side of said self-standing plastic sleeve, the two side sleeves connected via an intermediary plastic sheet; said water jacket sleeves configured to jointly envelop self-standing plastic sleeve and configured collectively with the self-standing plastic sleeve to be enclosed in said elongate translucent container.

14. The combination of claims 13, wherein said two water jacket sleeves with the connecting intermediary sheet form a self-standing jacket separable from the self-standing plastic sleeve.

15. The combination of claims 13, wherein said two water jacket sleeves are inside the self-standing plastic sleeve formed from sealed gussets in said self-standing plastic sleeve, in positions not interfering with the operation of said sparger tubes.

16. The combination of claims 13, wherein said two water jacket sleeves are further alternatingly inflated; said alternating inflation causing generation of waves in said bag medium.

17. The combination of claim 13, wherein said flowing fluid further containing electrochromic polymers or nanoparticles providing fluid color change of said jackets for biotuning and enhancing algae and cyanobacteria growth by pulsing electrical current at selected frequencies or for reducing photoinhibition effects from excess sunlight.

18. The combination of claim 13, wherein said two water jacket sleeves have jacket sleeve portions bonded on each side of said centrally-located self-standing plastic sleeve,

19. The combination of claims 1 and 4, further including a height-adjustable system to control medium level comprising lining said container provided with multiple recesses with said light transmitting plastic sheet; one of said recesses including a volume-expandable flexible translucent chamber; expanding said chamber volume causing said medium level to rise above all said recesses; retracting said chamber volume causing said medium to retrieve in the recess containing said retracted chamber; medium present in other recesses becomes isolated from other recesses and becomes available for processing.

20. The combination of claims 1 and 19, further including performing similar or dissimilar processes in self-standing plastic sleeve portions lining each separate recesses, such as, but not limited to containing highly porous matrices or gel, such as gel silica, for entrapping and milking cyanobacteria.

21. The combination of claims 19 and 20, wherein said processes are selected from the group consisting of exposing said medium to intense light of selected wavelength, to heat, to cold, to electromagnetic fields, to osmosis exchange, to catalysts, to cell density counting, to physical measurement, to continuous removal of secondary metabolites, to bio organic solvents, to electrocatalysts, to biocatalytic reductions, to fermentation, to dewatering, to harvesting, to extraction, to bacteria milking, to collection of secretion and to a combination thereof.

22. The combination of claims 1 to 21, wherein said bag is configured to line a surface selected from the group consisting of a flexible semi-rigid translucent fiberglass container provided with plastic memory container, a translucent container created by encasing a flat flexible semi-rigid plastic sheet into multiple bracket-type supports, a translucent profiled container positioned over a surface of same-container profile including recesses for receiving sunlight from all directions, embossed surfaces created by a soil compactor on stabilized earth, on stabilized clay, on cement, a translucent fiberglass container supported by buoys and floating over water, a translucent fiberglass container container to trusses of a greenhouse or of a warehouse, and a combination thereof.

23. The combination of claims 1 to 22, wherein a solar reflector film or a reflective mineral is positioned generally underneath said bag in a location such as under said semi-rigid container, bonded to the underside of said water jackets, or covering recess portions in said embossed surfaces.

24. The combination of claims 1 to 23, wherein a photobioreactor bag module is about 15m to 30m long (50 to 100 ft), is about lm wide layflat ( 40 inches) with a thickness of about 100µm to about 300µm (0.004 inches to about 0.012 inches)

25. The combination of claims 1 to 24, wherein each said sparger tubes is comprising a self-standing plastic sleeve portion having perforated holes being tucked into said sleeve similar to a gusset and pleats with two edge portions including four plastic sheet layers from said gusseted edges being bonded together.

26. A kit for culturing and agitating an algal medium, comprising:
a translucent, wide, semi-rigid elongate container, an elongate photobioreactor bag lining said container comprising a self-standing light transmitting plastic sleeve forming a closed chamber, the sleeve formed to include two parallel sparger tubes positioned at a location favorable to effect circulation of medium with bubbles alone;
a water jacket comprising a light transmitting tubular sleeve cooperating contactingly and heat exchangingly with said self-standing plastic sleeve;
a solar reflector film positioned under said container;
a three-way valve controller for diverting alternately incoming gases to said two sparger tubes;
and the bag further comprising at least three inlet ports for introducing gases or liquids and at least two exit ports for exhausting gases or removing liquids.

27. The kit of claim 26, wherein said water jacket sleeve is separable from said self-standing plastic sleeve.

28. The kit of claim 26, wherein said two water jacket sleeves are inside the self-standing plastic sleeve formed from sealed gussets in said self-standing plastic sleeve, in positions not interfering with the operation of said sparger tubes.

29. The kit of claim 26, wherein said two water jacket sleeves are further alternatingly inflated; said alternating inflation causing generation of waves in said bag medium.

30. The kit of claim 26, wherein said two water jacket sleeves have jacket sleeve portions bonded on each side of said centrally-located self-standing plastic sleeve.

31. The kit of claim 26, further including a height-adjustable system to control medium level comprising lining said container provided with multiple recesses with said light transmitting plastic sheet; one of said recesses including a volume-expandable flexible translucent chamber; expanding said chamber volume causing said medium level to rise above all said recesses; retracting said chamber volume causing said medium to retrieve in the recess containing said retracted chamber; medium present in other recesses becomes isolated from other recesses and becomes available for processing.

32. The kit of claim 31, further including performing similar or dissimilar processes in self-standing plastic sleeve portions lining each separate recesses, such as, but not limited to containing highly porous matrices or gel, such as gel silica, for entrapping and milking cyanobacteria.

33. The kit of claim 31, wherein said processes are selected from the group consisting of exposing said medium to intense light of selected wavelength, to heat, to cold, to electromagnetic fields, to osmosis exchange, to catalysts, to cell density counting, to physical measurement, to continuous removal of secondary metabolites, to bio organic solvents, to electrocatalysts, to biocatalytic reductions, to fermentation, to dewatering, to harvesting, to extraction, to bacteria milking, to collection of secretion and to a combination thereof.

34. The kit of claims 26 to 33, wherein said bag is configured to line a surface selected from the group consisting of a flexible semi-rigid translucent fiberglass container provided with plastic memory container, a translucent container created by encasing a flat flexible semi-rigid plastic sheet into multiple bracket-type supports, a translucent profiled container positioned over a surface of same-container profile including recesses for receiving sunlight from all directions, embossed surfaces created by a soil compactor on stabilized earth, on stabilized clay, on cement, a translucent fiberglass container supported by buoys and floating over water, a translucent fiberglass container container to trusses of a greenhouse or of a warehouse, and a combination thereof.

35. The kit of claim 26, further including elongate pivotable flaps, each radially movable by each of said sparger tubes thereby increasing amplitude of agitation in said medium.

36. A continuous method of inserting tangentially a fluid along the length of a closed flexible heat bondable sleeve without cutting open said sleeve; said method comprising first perforating apertures into an optionally flat or a folded sleeve portion, second tucking said perforated portion into said sleeve creating a gusset and third bonding the two pleated edges of said gusset forming an internal perforated tube inside said sleeve.

37. The continuous method of claim 26, wherein said perforated tube is a sparger tube.