U.S. patent application number 15/273887 was filed with the patent office on 2017-08-17 for methods and materials for cultivation and/or propagation of a photosynthetic organism.
The applicant listed for this patent is Forelight, Inc.. Invention is credited to Adam Flynn, Jeff Kantarek.
Application Number | 20170233689 15/273887 |
Document ID | / |
Family ID | 49161839 |
Filed Date | 2017-08-17 |
United States Patent
Application |
20170233689 |
Kind Code |
A1 |
Flynn; Adam ; et
al. |
August 17, 2017 |
METHODS AND MATERIALS FOR CULTIVATION AND/OR PROPAGATION OF A
PHOTOSYNTHETIC ORGANISM
Abstract
The present disclosure provides methods and materials for the
cultivation and/or propagation of a photosynthetic organism. Such
methods may comprise the use of a lamp assembly that comprises a
plurality of circuit boards, each comprising at least three edges,
arranged in a substantially spherical shape defining an interior
lamp assembly volume, wherein the plurality of circuit boards
comprise a first planar surface in contact with the interior lamp
assembly volume and an opposing second planar surface comprising
light emitting diodes (LEDs); and a barrier that surrounds the
plurality of circuit boards forming the substantially spherical
shape.
Inventors: |
Flynn; Adam; (Chicago,
IL) ; Kantarek; Jeff; (Chicago, IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Forelight, Inc. |
Chicago |
IL |
US |
|
|
Family ID: |
49161839 |
Appl. No.: |
15/273887 |
Filed: |
September 23, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14704516 |
May 5, 2015 |
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15273887 |
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13833079 |
Mar 15, 2013 |
9057043 |
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14704516 |
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61612001 |
Mar 16, 2012 |
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Current U.S.
Class: |
435/134 |
Current CPC
Class: |
C12M 23/02 20130101;
C12M 23/48 20130101; C12N 1/12 20130101; C12M 23/22 20130101; C12M
41/06 20130101; C12P 7/6427 20130101; C12P 39/00 20130101; C12M
41/24 20130101; C12M 41/48 20130101; C12M 21/02 20130101; C12P 7/62
20130101; C12M 31/08 20130101; C12M 31/10 20130101 |
International
Class: |
C12M 1/00 20060101
C12M001/00; C12P 7/64 20060101 C12P007/64; C12M 3/00 20060101
C12M003/00; C12N 1/12 20060101 C12N001/12; C12M 1/36 20060101
C12M001/36; C12M 1/02 20060101 C12M001/02 |
Claims
1. A photobioreactor for cultivation and/or propagation of a
photosynthetic organism comprising: a.) a vessel having a wall
defining an interior vessel volume; and b.) a lamp assembly
positioned within the interior vessel volume, wherein the lamp
assembly comprises: a plurality of circuit boards, each comprising
at least three edges, arranged in a substantially spherical shape
defining an interior lamp assembly volume, wherein the plurality of
circuit boards comprise a first planar surface in contact with the
interior lamp assembly volume and an opposing second planar surface
comprising light emitting diodes (LEDs); and a barrier that
surrounds the plurality of circuit boards forming the substantially
spherical shape.
2. The bioreactor of claim 1, wherein the vessel is cylindrical and
comprises a cylindrical wall, an upper wall, and a lower wall each
defining the interior tank volume.
3. The bioreactor of claim 1, wherein the vessel is substantially
spherical.
4. The bioreactor of claim 1, wherein the vessel comprises a hole
for a gas inlet, a hole for a gas outlet, or both.
5-12. (canceled)
13. The bioreactor of claim 1, wherein the lamp assembly comprises:
a plurality of circuit boards, each comprising at least three
edges, arranged in a substantially spherical shape defining an
interior lamp assembly volume, wherein the plurality of circuit
boards comprise a first planar surface in contact with the interior
lamp assembly volume and an opposing second planar surface
comprising light emitting diodes (LEDs); and a barrier that
surrounds the plurality of circuit boards forming the substantially
spherical shape.
14-27. (canceled)
28. A method of producing docosahexaenoic acid (DHA), the method
comprising: a.) providing one or more photosynthetic organisms
comprising enzymes for generating DHA; b.) adding the
photosynthetic organisms to a vessel of a bioreactor comprising a
liquid growth media; c.) contacting the one or more photosynthetic
organisms with light emitted from a lamp assembly, wherein the lamp
assembly comprises: a plurality of circuit boards, each comprising
at least three edges, arranged in a substantially spherical shape
defining an interior lamp assembly volume, wherein the plurality of
circuit boards comprise a first planar surface in contact with the
interior lamp assembly volume and an opposing second planar surface
comprising light emitting diodes (LEDs); and a barrier that
surrounds the plurality of circuit boards forming the substantially
spherical shape; and d.). producing DHA from the one or more
photosynthetic organisms.
29. The method of claim 28, wherein the one or more photosynthetic
organisms are algae.
30. A method for storage of a light energy, the method comprising
a.) providing one or more photosynthetic organisms comprising
enzymes for generating one or more compounds from a light energy;
b.) adding the one or more photosynthetic organisms to a tank of a
bioreactor comprising a liquid growth media; c.) contacting the one
or more photosynthetic organisms with light emitted from a lamp
assembly, wherein the lamp assembly comprises: a plurality of
circuit boards, each comprising at least three edges, arranged in a
substantially spherical shape defining an interior lamp assembly
volume, wherein the plurality of circuit boards comprise a first
planar surface in contact with the interior lamp assembly volume
and an opposing second planar surface comprising light emitting
diodes (LEDs); and a barrier that surrounds the plurality of
circuit boards forming the substantially spherical shape; and d.).
producing one or more compounds from the light energy.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 14/704,516 filed May 5, 2015, which is a
continuation of U.S. patent application Ser. No. 13/833,079 filed
Mar. 15, 2013 (now U.S. Pat. No. 9,057,043), which claims priority
to U.S. Provisional Application No. 61/612,001 filed Mar. 16,
2012.
BACKGROUND
[0002] Photobioreactors have been described for the use of
cultivating alga and generally employ shallow lagoons agitated with
one or more paddle wheels. Such bioreactors are plagued with
problems including poor production of algae due to seasonal and
daily climatic changes and contamination. Given that such
bioreactors are generally constructed to receive the sun's daylight
light, productivity is limited by intensity of the sun which
depends on the photoperiod and the season, among other factors.
SUMMARY
[0003] Methods and materials are provided for the cultivation
including, for example, propagation of a photosynthetic
organism.
[0004] The methods disclosed herein may comprise the use of a
photobioreactor that comprises the use of an electromagnetic source
in the visible light spectrum. In one embodiment, the
electromagnetic source comprises a plurality of circuit boards. In
another embodiment, each circuit board comprises at least three
edges, arranged in a substantially spherical shape defining an
interior lamp assembly volume. In another embodiment, the plurality
of circuit boards arranged in a substantially spherical shape
comprise a first planar surface in contact with the interior lamp
assembly volume and an opposing second planar surface comprising
high intensity lamps including, for example, light emitting diodes
(LEDs). In another embodiment, the photobioreactor comprises a
barrier that surrounds the plurality of circuit boards, said
barrier having a substantially spherical, ovoid, egg, cylindrical,
rectangular prismic or other similar shape. Such methods may be
used to produce compounds (e.g., biomolecules) including, for
example, fatty acids, phycobiliproteins such as C-Phycocyanin,
allophycocyanin, phycoerythrin, biofuels such as phytol, and other
various petrol fuel substitutes.
[0005] The present disclosure also provides a photobioreactor for
cultivation and/or propagation of a photosynthetic organism. In one
embodiment, the photobioreactor comprises an electromagnetic source
in the visible light spectrum. In another embodiment, the
photobioreactor comprises a vessel having a wall defining an
interior vessel volume. In one embodiment, the vessel is
substantially cylindrical, spherical, rectangular prismic or ovoid
in shape. In another embodiment, the photobioreactor comprises a
lamp assembly positioned within the interior vessel volume, wherein
the lamp assembly optionally comprises a plurality of circuit
boards, each optionally comprising at least three edges, arranged
in a substantially spherical or ovoid shape defining an interior
lamp assembly volume. In one embodiment, the plurality of circuit
boards each comprise a first planar surface in contact with the
interior lamp assembly volume and an opposing second planar surface
comprising light emitting diodes (LEDs). In one embodiment, the
photobioreactor comprises a barrier that surrounds the plurality of
circuit boards. In one embodiment, the barrier is substantially
cylindrical, spherical, rectangular prismic or ovoid in shape.
[0006] In some embodiments, which may be combined with any of the
above or below embodiments, the barrier is cylindrical and
comprises a cylindrical wall, an upper wall, and a lower wall each
defining an interior tank volume.
[0007] In some embodiments, which may be combined with any of the
above or below embodiments, the vessel is substantially spherical
or ovoid.
[0008] In some embodiments, which may be combined with any of the
above or below embodiments, the vessel comprises an opening for a
gas inlet.
[0009] In some embodiments, which may be combined with any of the
above or below embodiments, the vessel comprises an opening for a
gas outlet.
[0010] In some embodiments, which may be combined with any of the
above or below embodiments, the vessel comprises an opening for
wiring the light source.
[0011] In some embodiments, which may be combined with any of the
above or below embodiments, the lamp assembly is positioned
substantially in the center of the vessel.
[0012] In some embodiments, which may be combined with any of the
above or below embodiments, two or more lamp assemblies are
positioned in the vessel.
[0013] In some embodiments, which may be combined with any of the
above or below embodiments, the two or more lamp assemblies are
positioned at different heights in the vessel.
[0014] In some embodiments, which may be combined with any of the
above or below embodiments, three or more lamp assemblies are
positioned in the vessel.
[0015] In some embodiments, which may be combined with any of the
above or below embodiments, the three or more lamp assemblies are
positioned at different heights in the vessel.
[0016] In some embodiments, which may be combined with any of the
above or below embodiments, the three or more lamp assemblies are
positioned in a helical arrangement in the vessel.
[0017] The present disclosure also provides a light source for use
in cultivation and/or propagation of a photosynthetic organism. In
one embodiment, the light source comprises: a plurality of circuit
boards, each comprising at least three edges. In one embodiment,
the circuit boards are arranged in a substantially spherical shape
defining an interior lamp assembly volume, wherein the plurality of
circuit boards comprise a first planar surface in contact with the
interior lamp assembly volume and an opposing second planar surface
comprising light emitting diodes (LEDs); and a barrier that
surrounds the plurality of circuit boards forming the substantially
spherical shape.
[0018] In some embodiments, which may be combined with any of the
above or below embodiments, the substantially spherical shaped
arrangement of the planar circuit boards has a side devoid of at
least one circuit board to permit electrical connectivity.
Alternatively the spherical shaped arrangement of the planer
circuit boards has an aperture to permit electrical
connectivity.
[0019] In some embodiments, which may be combined with any of the
above or below embodiments, the circuit boards comprise two or more
tabs around their perimeter that form one or more notches that
permit the circuit boards to interlock.
[0020] In some embodiments, which may be combined with any of the
above or below embodiments, the circuit boards are pentagon
shaped.
[0021] In some embodiments, which may be combined with any of the
above or below embodiments, eleven pentagons are joined to form a
dodecahedron devoid of one side. In another embodiment, twelve
pentagons are joined together to form a dodecahedron with an
aperture in one or more of the pentagons.
[0022] In some embodiments, which may be combined with any of the
above or below embodiments, the circuit boards are triangular
shaped.
[0023] In some embodiments, which may be combined with any of the
above or below embodiments, twenty triangles are joined to form an
icosahedron devoid of one side. In another embodiment, twenty one
triangles are joined to form an icosahedron with an aperture in one
or more triangles.
[0024] In some embodiments, which may be combined with any of the
above or below embodiments, the circuit boards comprise red, white,
and blue LEDs.
[0025] In some embodiments, which may be combined with any of the
above or below embodiments, the red, white, and blue LEDs are
positioned adjacent to an LED of opposing color. In some
embodiments, which may be combined with any of the above or below
embodiments, the LEDs are pulse width modulated.
[0026] In some embodiments, which may be combined with any of the
above or below embodiments, the barrier is plastic.
[0027] In some embodiments, which may be combined with any of the
above or below embodiments, the barrier is substantially spherical
or ovoid.
[0028] In some embodiments, which may be combined with any of the
above or below embodiments, the plastic permits transmission of
light.
[0029] In some embodiments, which may be combined with any of the
above or below embodiments, the barrier has an opening to permit
electrical connectivity.
[0030] In some embodiments, which may be combined with any of the
above or below embodiments, a void between the barrier and the
circuit boards comprises a fluid for dispersal of heat.
[0031] In some embodiments, which may be combined with any of the
above or below embodiments, the fluid is mineral oil.
[0032] The present disclosure also provides methods of producing
docosahexaenoic acid (DHA) comprising: providing one or more
photosynthetic organisms comprising enzymes for generating DHA;
adding the photosynthetic organisms to a vessel of a bioreactor,
for example as described herein, comprising a liquid growth media;
contacting the one or more photosynthetic organisms with light
emitted from a lamp assembly. In one embodiment, the lamp assembly
comprises a plurality of circuit boards. In another embodiment,
each of the circuit boards are arranged in a substantially
spherical shape defining an interior lamp assembly volume, wherein
the plurality of circuit boards comprise a first planar surface in
contact with the interior lamp assembly volume and an opposing
second planar surface comprising light emitting diodes (LEDs). In
one embodiment, the lamp assembly comprises a barrier that
surrounds the plurality of circuit boards forming the substantially
spherical or ovoid shape. In other embodiments, the plurality of
circuit boards are arranged in a shape of any 4-, 6- or 8-sided
triangular, planar geometric shape such as a tetrahedron,
two-stacked tetrahedrons, an octahedron, or a 20 sided planar
geometric shape, such as an icosahedron.
[0033] In some embodiments, which may be combined with any of the
above or below embodiments, the one or more photosynthetic
organisms comprise algae and/or a productive algal culture.
[0034] In some embodiments, which may be combined with any of the
above or below embodiments, two or more algae are provided that
have natural environments that are similar in salinity and
dissimilar in temperature.
[0035] In some embodiments, which may be combined with any of the
above or below embodiments, the algae are selected from the group
consisting of Isochrysis aff. Galbana, pavlova lutheri, arthrospira
platensis, chiorella pyrenoidosa, synechococcus elongates,
including naturally occurring or genetically modified/recombinant
strains of the foregoing.
[0036] In some embodiments, which may be combined with any of the
above or below embodiments, the methods further comprise isolating
the DHA from the growth media.
[0037] The present disclosure also provides methods for storage of
a light energy comprising: providing one or more photosynthetic
organisms comprising enzymes for generating one or more compounds
from a light energy; adding the one or more photosynthetic
organisms to a tank of a bioreactor comprising a liquid growth
media; contacting the one or more photosynthetic organisms with
light emitted from a lamp assembly, for example as set forth
herein, and producing one or more compounds from the light energy.
In one embodiment, the lamp assembly comprises: a plurality of
circuit boards, each comprising at least three edges, arranged in a
substantially spherical shape defining an interior lamp assembly
volume, wherein the plurality of circuit boards comprise a first
planar surface in contact with the interior lamp assembly volume
and an opposing second planar surface comprising light emitting
diodes (LEDs); and a barrier that surrounds the plurality of
circuit boards forming the substantially spherical shape.
[0038] In some embodiments, which may be combined with any of the
above or below embodiments, energy is subsequently released from
the one or more compounds.
BRIEF DESCRIPTION OF THE DRAWINGS
[0039] The foregoing summary, as well as the following detailed
description of the disclosure, will be better understood when read
in conjunction with the appended figures. It should be understood
that the disclosure is not limited to the precise arrangements,
examples and instrumentalities shown.
[0040] FIG. 1 shows a schematic of an exemplary spherical-shaped
photobioreactor as described herein.
[0041] FIG. 2 shows a schematic of an exemplary dodecahedron shaped
light source comprised of pentagonal-shaped circuit boards (FIG.
2A), an exemplary icosahedron shaped light source comprised of
triangular-shaped circuit boards (FIG. 2B).
[0042] FIG. 3 shows a schematic of an exemplary lamp assembly
comprising a light source comprised of pentagonal-shaped circuit
boards (FIG. 3A), an exemplary lamp assembly comprising a light
source comprised of triangular-shaped circuit boards (FIG. 3B).
[0043] FIG. 4 shows a schematic of the interior of an exemplary
vertical barrel-shaped photobioreactor as described herein.
[0044] FIG. 5 shows a schematic of an exemplary vertical
barrel-shaped photobioreactor as described herein.
[0045] FIGS. 6A-6F show various positions of lamp assemblies within
a horizontal barrel shaped photobioreactor as described herein.
[0046] FIG. 7 shows an exemplary rack configuration for horizontal
barrel shaped photobioreactors as described herein.
[0047] FIGS. 8A-8B show exemplary rack connection configurations
for horizontal barrel shaped photobioreactors A and B as described
herein.
[0048] FIG. 9 shows a block diagram of an example control system
for managing the growth of cultures, according to an example
embodiment of the present invention.
[0049] FIG. 10 shows a flow diagram illustrating example procedures
to control the growth of a culture, according to an example
embodiment of the present invention.
[0050] FIGS. 11A-11B show an illustrative vessel lid (FIG. 11A) and
illustrative pebble configurations (FIG. 11B).
[0051] FIGS. 12 to 28 are diagrams showing embodiments of lamp
assemblies described herein.
DETAILED DESCRIPTION
[0052] The present disclosure provides methods and materials for
the cultivation and/or propagation of a photosynthetic organism
such as an alga using a photobioreactor. The photobioreactor
provided herein comprises a lamp assembly that comprises a
substantially spherical light source positioned in a vessel of the
photobioreactor that comprises a liquid medium and a photosynthetic
organism. The use of such a photobioreactor permits unexpectedly
high growth and density of the photosynthetic organism thus
maximizing bioconversion efficiencies and product yields from the
photosynthetic organism. The methods provided herein may be used to
produce one or more compounds (e.g., biomolecules) including but
not limited to fatty acids such as docosahexaenoic add (DHA),
docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA) or other
fatty acids or other compounds such as phycobiliproteins (e.g.
C-Phycocyanin, Allophycocyanin, Phycoerythrin, etc), and biofuels
such as phytol and other various petrol fuel substitutes) from the
photosynthetic organism. Additionally or alternatively, such
methods disclosed herein may be used to provide for storage of a
light energy.
[0053] Referring now to FIG. 1, a photobioreactor in accordance
with an embodiment of the present invention is shown.
Photobioreactor 101 comprises a vessel 102 (e.g., a tank) for
containing a liquid culture medium 103 for cultivating and/or
propagating a photosynthetic organism.
[0054] The photobioreactor 101 of the invention is suitable for the
culture of any kind of photosynthetic microorganism, such as a
plant cell and unicellular or multicellular microorganisms having a
light requirement. As used herein, the term "photosynthetic
microorganism" also includes organisms genetically modified by
techniques well known to one skilled in the art.
[0055] The liquid culture medium is sometimes referred to herein as
an "algal" culture, but it will be appreciated that the
photobioreactor may be employed for the cultivation of any type of
photosynthetic microorganism.
[0056] The vessel 102 may be covered by lid 104. In one embodiment,
lid 104 is constructed of an inert material including, for example,
plastics such as polyvinyl chloride, high-density polyethylene,
low-density polyethylene and polypropylene. The top rim of the
vessel 102 may be formed into a lip which permits lid 104 to be
secured (e.g., bolted or clamped) to the top of vessel 102. In
addition, the top rim of vessel may be fitted with a gasket
material to provide a liquid and gas-tight seal with lid 104. In an
embodiment, lid 104 may be lined with a reflective material. In an
embodiment, a rigid framing (e.g., metal framing) may be added to
solidify the vessel.
[0057] Vessel 102 may comprise a hole for a gas inlet 105. In an
embodiment, a portion of the gas inlet inside the vessel is capped
with an air stone or metal foam 107. Additionally, vessel 102 may
comprise gas vents 106 to permit exit of gas from the vessel.
[0058] The vessel 102 may be of any convenient shape, for example
substantially spherical or cylindrical. Vessel 102 may be made of
food grade or highly inert materials that do not leech and are
corrosion resistant including, for example, plastics such as
high-density polyethylene, low-density polyethylene and
polypropylene. Alternatively, vessel 102 may be constructed of
stainless steel, glass and the like. It is preferred that the
vessel be constructed of a heat resistant material and/or a
material that can withstand light pressurization.
[0059] A lamp assembly 108 (e.g. pebble) comprising light source
109, barrier 110, and an electrical connector 111 is suspended in
the interior vessel volume 112. In an embodiment, the portion of
the electrical connector 111 that resides inside the vessel is
waterproofed. In one embodiment, the light source comprises a
plurality of circuit boards 113, each comprising at least three
edges, at least 4 edges or at least 5 edges, arranged in a
substantially spherical shape defining an interior light source
volume 114, wherein the plurality of circuit boards comprise a
first planar surface 115 in contact with the interior light source
volume and an opposing second planar surface 116 comprising high
intensity lamps such as light emitting diodes (LEDs) 117 with an
emission spectrum suitable for the growth of a photosynthetic
organism; and a barrier 110 that surrounds the plurality of circuit
boards forming the substantially spherical shape. Alternatively, a
single circuit board (e.g., a flexible board) may be molded into a
substantially spherical shape and used in the light assembly.
[0060] The circuit boards 116 may comprise red, white, and blue
LEDs. Such LEDs may be positioned in groups containing two or more
LEDs of the same color positioned adjacent to two or more LEDs of
another color. Alternatively, an LED including, for example, a red,
white, or blue LED, may be positioned adjacent to an LED of
opposing color. In some embodiments, the LEDs may be positioned in
rows such that a single LED is adjacent to four or six LEDs. The
LEDs may emit light of uniform intensity or of varying
intensity.
[0061] The substantially spherical shaped arrangement of the planar
circuit boards may have an opening or a side devoid of at least one
circuit board to permit electrical connectivity.
[0062] The circuit board may be of any polygonal shape. In one
embodiment, the polygonal shape permits several circuit boards to
be joined into a substantially spherical shape with an interior
volume. In some embodiments, the circuit boards may be pentagon
shaped. In a further embodiment, eleven pentagons are joined to
form a dodecahedron devoid of one side (FIG. 2A). Alternatively, in
some embodiments, the circuit boards may be triangular in shape. In
a further embodiment, twenty triangles are joined to form an
icosahedron devoid of one side (FIG. 2B). In another embodiment,
six triangles are joined to form a double pyramid with a triangular
base (FIG. 2C). In another embodiment, eight triangles are joined
to form a double pyramid with a square base (FIG. 2D).
[0063] The circuit boards may also comprise a copper or other metal
layer that faces the interior volume for dissipation of heat from
the lamps (e.g., LEDs) or signaling.
[0064] In some embodiments, the circuit boards comprise two or more
tabs around their perimeter that form one or more notches that
permit the circuit boards to interlock (see for example FIG. 3A
(triangle) and FIG. 3B (pentagon).
[0065] The barrier 110 surrounding the plurality of circuit boards
forming the substantially spherical shape may be constructed of a
variety of inert materials, such as various plastic materials
including, for example, transparent plastic and/or plastic tolerant
to extreme temperatures. The barrier functions to provide a
water-tight seal around the plurality of circuit boards to prevent
a culture medium in the interior vessel volume 112 coming in
contact with the plurality of circuit boards 116. In some
embodiments, the barrier 110 surrounding the plurality of circuit
boards forms a container (e.g., a jar) around the circuit boards
which may be substantially cylindrical (FIG. 3A) or spherical (FIG.
3B) in shape. In some embodiments, the container formed by the
barrier holds one or more objects to reduce the buoyancy of the
light source (e.g., inert beads such as glass beads 119).
[0066] The circuit board may be populated with LEDs that emit light
of one or more wavelengths to optimize for a particular strain of
photosynthetic life or to express a specific feature for abnormal
growth. In some embodiments, the circuit boards may be populated
with lamps that emit electromagnetic radiation outside the visible
spectrum including, for example, UV and/or IR light. Such lamps may
be used to sterilize the liquid media in the vessel (e.g., lamps
emitting UV light) and/or heat the liquid media in the vessel
(e.g., lamps emitting IR light). In some embodiments, the LEDs may
be pulse width modulated. In one embodiment, the pulse width
modulation is optimized to maximize growth of the particular
microorganism in the photobioreactor. In another embodiment, the
duty cycle is at least 50%, at least 55%, at least 50%, at least
55%, at least 60%, at least 65%, at least 70%, at least 75%, at
least 80%, at least 85%, at least 90%, or at least 95%.
[0067] The circuit board may comprise sensors for collecting data
from the interior of the vessel. For example, sensors may be
configured to collect data on the culture media including, for
example, optical density, pH, and/or conductivity.
[0068] The plurality of circuit boards forming a substantially
spherical shape with an interior volume may comprise one or more
microprocessors in the interior volume including for individual
control of LEDs or control of banks of LEDS (e.g., two or more
LEDs) and/or feedback monitoring.
[0069] Vessel 102 may comprise one or more holes or access ports
for providing electrical connectivity to the lamp assembly 108.
[0070] One or more lamp assemblies 108 may be distributed
throughout the interior vessel volume 112. A single light source
may be suspended at the center of the interior vessel volume.
Alternatively, when two or more lamp assemblies are used in the
photobioreactor, the lamp assemblies may be suspended at the same
or different heights in the interior vessel volume (see FIGS. 1 and
4). Alternatively, when three or more lamp assemblies are used, the
lamp assemblies may be suspended in any geometric arrangement such
as a helical or double helical arrangement.
[0071] A cooling device may be provided to control the temperature
of the vessel. Such cooling device may be in the form of a cooling
jacket surrounding a portion of the wall of vessel 102. Such
cooling jacket provides circulating cooling water or other fluid
across the wall of vessel 102 to absorb heat and assist in
controlling the temperature of culture medium 103 contained in the
vessel. Cooling water may enter jacket through an inlet tube and
exit through an exit tube. The dimensions of the cooling jacket
will depend upon a number of factors, such as the amount of heat
transmitted by lamp assemblies 108 to the liquid culture medium
contained in vessel 102, the desired temperature of the culture
medium, the temperature and flow rate of the cooling water, and the
like. Alternatively, temperature of the vessel may be regulated via
temperature regulation of the lamp assemblies or LEDs, for example
by circulating and/or controlling temperature of an oil in which
the LEDs or lamp assembly is immersed.
[0072] Vessel 102 is generally designed to accommodate a head space
between the liquid culture surface and lid. The head space allows
for foaming, which often occurs in biological culture media.
[0073] Vessel 102 may be fitted with gas inlet tube 105 which is
provided with a pressurized gas (e.g., carbon dioxide or carbon
dioxide-enriched air) for supporting the photosynthetic
requirements of the algal culture. Tube 105 passes through the wall
of vessel 102. Gas bubbles rise through the liquid algal culture
medium contained in vessel 102 and the spent gases escape through
gas vents 106 which may be disposed in the wall or lid of vessel,
preferably above the surface level of the culture medium.
[0074] If more vigorous mixing is desired, an air pump 118 or other
agitation mechanism, for example powered by a motor, may be used to
agitate the culture medium.
[0075] The photobioreactor 101 may further comprise a cleaning unit
mounted within the interior vessel volume or on the outside surface
of the lamp assembly 108 for cleaning the outer surface of the lamp
assembly, and a cleaning unit actuator for actuating the cleaning
unit. Such a cleaning unit may function to get rid of the
cultivated and/or propagated photosynthetic organisms which may
block the light source by adhering to the outer lamp assembly. In
some embodiments, the vessel is composed of or coated with a
superhydrophobic, hydrophilic, and/and or oleophobic material.
[0076] Referring now to FIG. 5, this figure shows another
embodiment of a photobioreactor as provided by the instant
disclosure. Such a photobioreactor may comprise a cylindrical
shaped vessel 501 (e.g., a barrel shaped vessel) to permit stacking
or racking of the two or more vessels in a vertical or horizontal
position. Because of their general availability, cost, and ease of
storage (e.g., stackability), 55 gallon plastic drums can be
employed with the methods disclosed herein. In an embodiment, the
vessel 501 may be lined with a reflective material. In an
embodiment, the vessel comprises a lid 502 with gas exhaust
fittings 503, a CO.sub.2 injection line 504, and electrical inlet
fittings 505. In an embodiment, the lid may be attached to the
vessel via an attachment mechanism such as a clamp 506. The
electrical inlet fittings 505 may receive waterproofed electrical
lines 507 powered by power source 508. In some embodiments, the
vessel may comprise air line bulkhead fittings 509 and water line
bulkhead fittings 510.
[0077] Although the embodiment shown in FIG. 5 is depicted in a
vertical configuration, a photobioreactor consistent with the
instant disclosure may also be arranged in a horizontal
configuration. Such a photobioreactor may comprise a cylindrical
shaped vessel (e.g., a barrel shaped vessel) to permit stacking or
racking of the two or more vessels in a horizontal position. In one
embodiment, the vessel comprises a lid for introducing medium or
removing culture and a drain for introducing medium or removing
culture. In another embodiment, the photobioreactor comprises a gas
inlet positioned on the bottom of the photobioreactor. In one
embodiment, this gas inlet is a CO.sub.2 inlet. In another
embodiment, the photobioreactor comprises one to a plurality (e.g.
1 to about 50, 2 to about 30, 3 to about 20 or about 6, 8, 10 or
12) bottom gas fittings and one to a plurality (e.g. 1 to about 50,
2 to about 30, 3 to about 20 or about 6, 8, 10 or 12) of top gas
fittings. In one embodiment, as viewed from the base end of the
vessel with the drain at the bottom, the bottom gas fittings are
located substantially between about the 3- and about the 6-o'clock
positions, for example about the 3-, about the 3:30-, about the 4-,
about the 4:30-, about the 5-, about the 5:30- or about the
6-o'clock positions. In one embodiment, as viewed from the base end
of the vessel with the drain at the bottom, the top gas fittings
are located substantially between about the 9- and the 12-o'clock
positions, for example about the 9-, about the 9:30-, about the
10-, about the 10:30-, about the 11-, about the 11:30- or about the
12-o'clock positions. In one embodiment, air is circulated in and
out of the vessel via the bottom gas fittings and the top gas
fittings. In one embodiment, gas enters the vessel via the bottom
gas fittings and exits the vessel via to gas fittings. In one
embodiment, the vessel contains volume markers, for example gallon
markers.
[0078] The photobioreactor may further comprise a transferring
mechanism for transferring liquid media, nutrients and/or
antibacterial agents to the interior vessel volume. Nutrients may
include synthetic and/or organic nutrients. In some embodiments, an
anti-bacterial agent (e.g., a detergent) may be added to the liquid
media to slow, or prevent, the growth of contagens (e.g., any
organism that is other than the photosynthetic microorganism
purposefully added to the vessel) in the vessel. Nutrients and/or
antibacterial agents may be added to the vessel by any method known
in the art including, for example, via an automated drip system. In
an embodiment, the automated drip system may be connected (e.g.,
wired or wireless connection) to a monitoring system that monitors
nutrient levels in the vessel. Such a monitoring system may
constantly or periodically monitor nutrient levels in the vessel
and prompt the automated drip system to release nutrients into the
vessel when nutrient levels fall below a predetermined limit.
Conversely, the monitoring system may prompt the automated drip
system to stop the release of nutrients into the vessel when
nutrient levels exceed a predetermined limit. In some embodiments,
other trace chemicals (e.g., citric acid) may be added to the
vessel to optimize environmental conditions for the growth of the
photosynthetic microorganism.
[0079] FIGS. 6A to 6F show embodiments of lamp assemblies 108
included within a vessel 102. In each of these embodiments, the
lamp assemblies 108 are configured to be interconnected in series
along one or more connection lines 602. This connection scheme
enables power to be provided to more than one lamp assembly 108
within a vessel. The configurations of the light assemblies 108
within the vessels 102 are based on the strain in question and the
targeted product. For example, certain strains may have better
growth rates when only one connection line 602 is disposed within a
vessel while other strains have better growth rates when more than
one connection line is disposed within a vessel.
[0080] While FIGS. 6A to 6F show the connections being made in
series, the lamp assemblies 108 may include parallel electrical
connections so that a failure of one lamp assembly does not affect
the operation of other interconnected lamp assemblies. Further,
while the connection lines 602 are shown as being substantially
straight through the vessels 102, in other embodiments, the
connection lines 602 may be formed at one or more angles (e.g.,
lamp assemblies 108 may be connected at a 90 degree angle).
[0081] The example connection line 602 is configured to provide at
least one of power and immersion oil to each of the lamp assemblies
108. For example, in FIG. 6A the vessel 102a includes connection
line 602a, which includes ports 604a and 604b. The immersion oil is
provided to the connection line 602a through port 604a and exits
the connection line 602a at port 604b. In this manner, an operator
can cycle immersion oil through the lamp assemblies 108 connected
to the connection line 602a to control temperature. In addition,
electrical connector 111 is provided to each of the lamp assemblies
108 through port 604a. The control of power and oil to the lamp
assemblies 108 is described in further detail in conjunction with
FIGS. 9 and 10.
[0082] FIG. 6C shows that connection lines 602 may interconnect.
This interconnection facilitates the flow of immersion oil through
the vessel 102. Additionally or alternatively, the interconnection
may enable lamp assemblies 108 located at intersection points to be
controlled by either of the control signals on the intersecting
electrical connectors 111. For instance, the lamp assembly 108c may
be connected to electrical connectors 111 positioned within
respective ports 604c, 604d, and 604e. As a result, the lamp
assembly 108c may be controlled by a control signal provided on any
one of the electrical connectors 111.
[0083] FIG. 6D shows a connection line 602 with a lamp assembly 108
that includes twenty circuit boards with LEDs. In this embodiment,
immersion oil is cycled though the lamp assembly 108 to control
temperature. Further, the configuration of the twenty circuit
boards enables light to be directed to substantially any location
within the vessel 102. This is in comparison to the light
assemblies 108 of FIGS. 6A, 6B, 6C, 6E, and 6F that include
relatively fewer circuit boards. These figures accordingly use more
light assemblies to compensate for each light assembly having fewer
LEDs.
[0084] A large scale photobioreactor is also contemplated by the
present disclosure. Such a bioreactor may comprise one or more
vessels and one, two or more lamp assemblies. The vessels may be of
uniform volume and dimension or may be of varying volume and
dimension. For example, vessels that hold 55 gallons or greater, or
vessels that hold 3,000 gallons or greater, may be used in the
large scale photobioreactors disclosed herein. In an embodiment,
area may be provided between vessels to provide for connectivity of
one vessel to another. In another embodiment, the vessel may have a
central column for light distribution and/or electrical
connectivity.
[0085] It will be appreciated that the photobioreactors of the
invention can be produced in widely varying sizes. Several
photobioreactors may be grouped together to produce large scale
photobioreactors for example as shown in FIGS. 7 and 8. For
simplicity, a single photobioreactor employing one light assembly
has been illustrated in FIG. 1, however, in a typical industrial
scale photobioreactor, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50,
60, 70, 80, 90, 100 or more photobioreactors each comprising one or
more lamp assemblies may be used.
[0086] A photobioreactor with two or more vessels may comprise air
and water lines that connect a vessel with one or more adjacent
vessels. The air and water lines may comprise valves to direct the
flow of air and/or water in a predetermined manner. Optionally, the
valves may permit for quick disconnection of a vessel for
maintenance and/or inspection. FIGS. 7 and 8 show diagrams of
photobioreactor embodiments including multiple interconnected
vessels. In particular, FIG. 7 shows a photobioreactor embodiment
with multiple columns of vessels wherein each column of vessels is
interconnected to another column of vessels. FIG. 8 shows a
photobioreactor embodiment wherein multiple vessels in a single
column are interconnected.
[0087] In the illustrated embodiment of FIGS. 7 and 8, air enters
in through the twelve gas inlets at the 7:30 and 4:30 positions on
the barrel where the stream is broken up into a column of two
micron bubbles, which mix with the culture medium, adding
circulation and necessary gases. For strains with lower gas
requirements it is possible to subdivide the gas inlets using a
fraction of the inlets for gas and leaving the remainder of the
inlets unused or connected to a water recirculation pump. In this
embodiment, a gas bubble builds at the top of the lower barrels
until the gas reaches the 11:30 and 12:30 positions of the gas
outlet. At this point, the pressure of the bubble drives the same
process in the barrel above until the gas reaches the top of the
column. It should also be noted that the bottom barrel of each
column includes between two and four additional gas inlets coupled
to 0.5 micron diffusers, which allow for the utilization of more
exotic gases for gross supplementation (i.e. CO.sub.2, gaseous
ammonia, etc.).
[0088] In some embodiments, a large scale photobioreactor may
comprise a single injection point for air and/or nutrients such
that the introduction of air (e.g., CO.sub.2) and/or nutrients into
a first vessel operably connected with other vessels allows the air
and/or nutrients to move from the first vessel into the other
vessels.
[0089] The vessels of a large scale photobioreactor may be operably
connected to a control unit (e.g., a controller or a server) and/or
sensor monitoring unit. In some embodiments, the control unit may
be configured to permit an operator to control the lamp assembly in
each vessel (e.g., turn the lamp assembly on or off, or adjust the
intensity of the lamps). In some embodiments, the sensor unit may
be configured to disconnect a vessel from other vessels in the
photobioreactor upon sensing that the vessel has been
contaminated.
[0090] A high level block diagram of an example control system 900
is illustrated in FIG. 9. The example control system 900 includes a
workstation 902, a controller 904 (e.g., a control unit), a vessel
102, pumps 906, and an immersion oil container 908. While FIG. 9
shows the controller 904 providing control for only the vessel 102,
in other embodiments the controller 904 may be communicatively
coupled to two or more vessels. Further, the workstation 902 may be
communicatively coupled to more than one controller.
[0091] The example workstation 902 is configured to provide an
operator interface for providing instructions to the controller
904, controlling of the process, and displaying system process
data. The example controller 904 manages the control of lamp
assemblies 108 and the flow of immersion oil. In other examples,
the controller 904 can be configured to manage the control of the
liquid culture medium 103 within the vessel 102. For instance, the
controller 904 may be configured to control gas inlet valves, gas
vents, fluid inlet valves, and/or additive valves.
[0092] The example workstation 902 operates in conjunction with the
controller 904 to provide control and system feedback to an
operator. The workstation 902 includes any type of processor
including, for example, a personal computer, a laptop, a server, a
smartphone, a tablet computer, etc. The controller 904 includes any
type of control system (e.g., an Arduino.TM. microcontroller or an
Application Specific Integrated Circuit ("ASIC")) that is
configured to provide open or closed loop system control using
inputs from sensors to control lighting and oil flow.
[0093] In this example, the controller 904 is separately
electrically connected to lamp assemblies 108a-d via respective
electrical connectors 111a and 111b. The electrical connectors 111
are disposed within respective connection lines 602a and 602b. The
controller 904 uses the separate connection lines 602 to provide
separate control to the lamp assemblies 108 within the respective
connection lines 602. For example, the controller 904 may provide
power to the lamp assemblies 108a and 108b while placing lamp
assemblies 108c and 108d into an off state.
[0094] The example controller 904 is communicatively coupled to the
pumps 906. The controller 906 may provide digital instructions or
an analog PWM voltage to operate the pumps 906. In this example,
the system 900 includes a separate pump 906 for each of the
connection lines 602 so that the controller 904 can independently
control the flow of immersion oil to the respective lamp assemblies
108. In other embodiments, a single pump 906 is used to provide oil
to one or more connection lines 602.
[0095] The example pumps 906 include any type of component for
providing immersion oil to the connection lines 602. For example,
the pumps 906 can include displacement pumps, gear pumps, screw
pumps, roots-type pumps, peristaltic pumps, plunger pumps,
hydraulic pumps, velocity pumps, etc. Responsive to receiving a
control signal (e.g., a voltage) from the controller 904, the pumps
906 move immersion oil from the container 908 to the respective
connection lines 602. The pumps 906 can be configured to pump oil
at varying velocities. Alternatively, the pumps 906 may be
configured to operate in a binary state (e.g., On/Off). While the
pumps 906 are shown as being located at the entrance of the
connection lines 602, in other embodiments the pumps 906 may be
located at the exit of the connection lines 602. In these other
embodiments, the pumps 906 are configured to pull immersion oil
through the connection lines 602.
[0096] The example immersion oil container 908 may be any type of
tank to store immersion oil. In some examples, the container 908
may include a jacket that is positioned along a portion of the
vessel 102. Further, the oil container 908 may include a component
that provides active cooling or heating.
[0097] In the illustrated example of FIG. 9, the controller 904 is
configured to control a voltage, frequency, and PWM (e.g., duty
cycle) of power applied to the lamp assemblies 108. For example,
the controller 904 is configured to provide a voltage between 8.8
and 12 volts to each of the lamp assemblies 108. In other examples,
the controller 904 may be configured to provide a voltage between 0
and 24 volts to the lamp assemblies 108. The applied voltage is
used to control the intensity of light transmitted from the lamp
assemblies 108. In examples where the LEDs (or other light sources)
emit light proportional to the applied voltage, the controller 904
is configured to provide a voltage such that the LEDs emit light at
a light intensity optimal for growth of the current culture.
[0098] The frequency of light emitted by the LEDs may be in the
range between 390 to 700 nm. The frequency of light is based on the
type of light source. In some examples, the controller 904 is
configured to select which LEDs are activated to achieve the
desired light frequency. For instance, each of the lamp assemblies
108 may include red, green, and blue LEDs. The lamp assemblies 108
may also include one or more mirrors to combine the light
transmitted from the LEDs. The controller 904 may be configured to
time the pulsing of the different colored LEDs to achieve a
resulting light frequency that is optimal for the target culture.
Alternatively, light filters may be applied to the lamp assemblies
108 to achieve the desired light frequency.
[0099] In addition to controlling the magnitude of the voltage, the
controller 904 is also configured to cycle the voltage at a
specified PWM. For example, the controller 904 may cycle the lamp
assemblies 108 over a 100 millisecond (ms) time period such that
the LEDs are on for 75% of the time (e.g., 75 ms) and off for 25%
of the time. The controller 904 is accordingly programmed (or
provided instructions) for a duty cycle and time period. While a
75% duty cycle and 100 ms time period was used as an example, the
controller 904 can be programmed to operate the lamp assemblies 108
using any duty cycle and time period.
[0100] The controller 904 is also configured to control the flow of
immersion oil through the lamp assemblies 108 to regulate the
temperature within the vessel 102. The controller 904 uses
instructions provided from the workstation 902 to determine how the
power and oil flow is to be controlled. It should be appreciated
that different types of cultures have optimal growth settings and
as a result, the controller 904 can be programmed based on the
culture to be grown within the vessel 102.
[0101] The example controller 904 is also configured to monitor
conditions within the vessel 102 and report these conditions to the
workstation 902. In this embodiment, the vessel 102 includes
temperature sensors 912, which are communicatively coupled to the
controller 904. The controller 904 records temperature data
provided by the sensors 912 and periodically transmits the
temperatures to the workstation 902. The controller 904 is also
configured to report current operating conditions including, for
example, voltage, frequency, and PWM applied to each lamp assembly,
a duration of operation, and/or detected diagnostic faults within
the system 900 (e.g., a broken lamp assembly or obstruction within
a connection line 602), communication interference with the
workstation 902, etc.
[0102] FIG. 10 shows a flow diagram illustrating example procedures
1000 and 1050 to manage the growth of a culture within the vessel
102 of FIG. 9, according to an example embodiment of the present
invention. The example procedures 1000 and 1050 may be carried out
by, for example, the workstation 902, the controller 904, the pumps
906, and/or the lamp assemblies 108 described in conjunction with
FIG. 9. Although the procedures 1000 and 1050 are described with
reference to the flow diagram illustrated in FIG. 10, it will be
appreciated that many other methods of performing the acts
associated with the procedures 1000 and 1050 may be used. For
example, the order of many of the blocks may be changed, certain
blocks may be combined with other blocks, and many of the blocks
described may be optional.
[0103] It will be appreciated that all of the disclosed procedures
described herein can be implemented using one or more computer
programs or components. These components may be provided as a
series of computer instructions on any conventional
computer-readable medium, including RAM, ROM, flash memory,
magnetic or optical disks, optical memory, or other storage media.
The instructions may be configured to be executed by a processor
(e.g., the workstation 902 and/or the controller 904), which when
executing the series of computer instructions performs or
facilitates the performance of all or part of the disclosed methods
and procedures.
[0104] The example procedure 1000 begins when an operator uses the
workstation 902 to program a routine for a session to grow a new
culture (e.g., a photosynthetic algae culture) (block 1002). The
operator specifies a time period of 50 ms and a duty cycle of 55%.
The operator also specifies that the voltage applied to the LEDs is
to be 10.75 volts to generate a desired light intensity. The
operator further specifies that the culture is to be grown over a
two day period and that the temperature of the medium 103 is not to
exceed 105.degree. C. The operator moreover selects to receive
alerts when any issues are detected with any of the lamp assemblies
108 and a report that graphs the temperature of the vessel 102 over
time.
[0105] After programming the routine, the operator instructs the
workstation 902 to transmit instructions 1003 (e.g., parameters) to
the controller 904 (block 1004). The instructions 1003 include the
programmed routine and may be formatted in a programming language
compatible with the controller 904. The workstation 902 then begins
to receive and store process data 1005 from the controller 904
(block 1006). The process data 1005 includes operational and
diagnostic information indicative of the process at the vessel 102.
The workstation 902 may receive a relatively constant stream of
process data 1005 as the data is processed and transmitted by the
controller 904. Alternatively, the workstation 902 may periodically
receive the process data 1005 from the controller 904.
[0106] The workstation 902 analyzes the process data 1005 for any
alerts (block 1008). Additionally or alternatively, the workstation
902 determines whether an alert message 1007 was received from the
controller 904. If an alert was received, the workstation 902
notifies an operator of the alert (block 1010). The notification
can include, for example, a text message, an e-mail, an audio
message, etc. indicating the contents of the alert. In the
illustrated example, an alert can include an indication that a
temperature of the vessel 102 is greater than the 105.degree. C.
threshold. Alternatively, the alert can include information
indicating that one or more of the lamp assemblies are inoperable.
Moreover, the alert can include information indicating that either
of the pumps 906 is not performing as expected.
[0107] In the illustrated example of FIG. 10, the workstation 902
determines whether the session has expired after providing an alert
to the operator (block 1012). In other embodiments, the workstation
902 may instruct the controller 904 to pause the process until an
operator responds to the alert. In yet other embodiments, the
workstation 902 may provide the controller 904 instructions for
responding to an alert. For instance, after detecting that the lamp
assembly 108b is inoperative, the workstation 902 provides the
controller 904 instructions changing the duty cycle, time period,
and voltage for the other lamp assemblies 108a, 108c, and 108d to
compensate for the loss of light.
[0108] Responsive to not receiving an alert, the workstation 902
determines whether the session (e.g., the programmed two day
operating time) has ended (block 1012). Responsive to determining
the session has ended, the workstation 902 receives a report 1013
that includes the monitored temperature of the vessel 102 over the
two day period (block 1014). The example procedure 1000 then ends.
Alternatively, the example procedure 1000 returns to block 1002 for
the next session. However, if the workstation 902 determines that
the session has not ended (block 1012), the workstation 902 returns
to receiving process data 1005 from the controller (block 1006).
The example procedure 1000 then continues until the session
expires.
[0109] The example procedure 1050 begins when the controller 904
receives operation instructions 1003 from the workstation 902
(block 1052). The example controller 904 then configures its
operation to provide power to lamp assemblies 108 based on the
instructions 1003 (block 1054). The configuration can include
setting parameters for output drives so that the lamp assemblies
108 receive 10.75 volts at a 55% duty cycle of a 50 ms time period.
The example controller 904 also configures its connection to the
pumps 906. This configuration can include operating the pumps 906
so that there is sufficient immersion oil within the lamp
assemblies 108.
[0110] The example controller 904 also configures operation
conditions including, for example, an allowable temperature ranges
for the vessel 102, alert threshold temperatures for the vessel
102, diagnostic settings, etc. (block 1056). The controller 904
also determines which process data is to be transmitted to the
workstation 902 and which data is to be included within a report.
The configuration can further include calibration of the
temperature sensors 912 and/or the lamp assemblies 108.
[0111] After configuration, the controller 904 operates the lamp
assemblies at the specified voltage and duty cycle (block 1058).
The example controller 904 also begins receiving outputs from the
temperature sensors 912. For instance, temperature sensor 912a
reports the temperature in the liquid medium culture 103,
temperature sensor 912b reports the temperature within the lamp
assembly 108a, and temperature sensor 912c reports the temperature
of connection line 602b. It should be appreciated that in other
examples, only one of the temperature sensors 912 may be used. It
should also be appreciated that in other examples, the controller
904 may be configured to receive outputs from other types of
sensors (e.g., pH sensors, salinity sensors, light sensors,
chemical sensors, etc.
[0112] In this example, the controller 904 transmits the
temperature data as the process data (e.g., operational data) 1005
(block 1060). The process data 1005 can also include any faults
detected within the vessel 102 and/or within the controller 904.
The process data 1005 can further include the voltage and duty
cycle applied to the lamp assemblies 108 and whether the pumps 906
are being operated.
[0113] The example controller 904 then determines whether the
session should end (e.g., the two day period) (block 1062).
Responsive to determining that the session should end, the example
controller 904 compiles collected data, generates a report, and
transmits the report 1013 to the workstation 902 (block 1064). In
other embodiments, the example controller 904 ends the session and
notifies the workstation 902 that the session has ended without
providing a report. In these other embodiments, the controller 904
may not have the capability of generating a report. Instead, the
controller 904 may provide a log or data structure of collected
data at the end of the session. At the end of the session, the
example procedure 1050 ends. Alternatively, the example procedure
1050 returns to block 1052 to begin a session for another
culture.
[0114] Returning to block 1062, if the session has not ended, the
controller 904 compares the temperature outputs from the sensors
912 to pre-specified temperatures (block 1066). The controller 904
performs this comparison to determine whether action should be
taken to actively change the temperature within the vessel 102. For
example, if the temperature is outside of a specified allowable
range, the controller 904 operates the pumps 906 to circulate
cooler (or warmer) oil through the vessel 102 (block 1068). The
controller 904 continues to operate the pumps 906 until the
temperature is within the allowable range. Additionally or
alternatively, the controller 904 may also change the intensity of
the lamp assemblies 108 and/or the duty cycle to modify the
temperature. For instance, lowering the voltage applied to the lamp
assemblies 108 reduces the amount of heat transmitted to the vessel
102. Moreover, changing the duty cycle to have more time in an
`Off` state also reduces the amount of heat transmitted to the
vessel 102.
[0115] The example controller 904 also determines whether the
temperature of the vessel 102 exceeds a threshold (block 1070).
Responsive to determining the temperature exceeds a threshold, the
controller 904 transmits an alert message 1007 to the workstation
902 (block 1072). The controller 904 continues to operate the lamp
assemblies 108 until further instruction is provided by the
workstation 102 or until the session ends.
[0116] A photosynthetic microorganism such as an alga may be
cultivated and/or propagated with the photobioreactor as disclosed
herein. Generally, prior to filling the interior volume of the
vessel of the photobioreactor with a culture medium, the interior
of the photobioreactor is sanitized by exposing it to a sterilizing
gas, a hypochlorite solution or the like. Following sanitization,
water is introduced into the vessel via a pressurized water line.
The vessel is filled with water to a predetermined depth. In some
embodiments, the vessel is filed such that the surface of the
culture medium will be below an exit port. Nutrients and inocula
(e.g., one or more photosynthetic organisms such as an alga) are
introduced into the vessel by removing the lid or introducing them
through an access port in the vessel or lid. The pH and temperature
of the medium may be monitored throughout the photobioreaction by a
temperature probe and a pH probe. The photobioreaction is initiated
by supplying electrical power to the lamp assembly and optionally
initiating sparging of an appropriate gas mixture via an inlet
tube. The progress of the photobioreaction may be monitored with a
calibrated density detector. Adjustments to the pH or the
composition of the culture medium may be effected by introducing
materials through the lid or an access port in the vessel or
lid.
[0117] In some embodiments, a starter culture of each
photosynthetic microorganism to be added to the vessel may be
grown. After the starter culture reaches an optimal density, a
portion of the culture may be added to the vessel. In some
embodiments, where two or more types (e.g., species or strains) of
microorganisms are added to the vessel, an equal number of each
type of photosynthetic microorganism may be added to the vessel.
Alternatively, an unequal number of each strain of photosynthetic
microorganism may be added to the vessel.
[0118] When the photosynthetic microorganisms are ready to harvest,
the light source and optional sparging gas are turned off and the
medium containing the photosynthetic microorganisms is collected.
In an embodiment, the medium and photosynthetic microorganism may
be collected via the opening of an optional valve located on vessel
wall. In a further embodiment, a pump may be used to expel the
medium and photosynthetic microorganism from the vessel.
Alternatively, in embodiments where the vessel comprises a gas
inlet (e.g., for CO.sub.2 injection), the gas inlet may be reversed
to allow static pressure to build up and force the media out via an
alternate line. Alternatively, the photosynthetic microorganism may
agglomerate and be expelled from the vessel (e.g., by excess air
flow) onto a surface external to and apart from the vessel. In
other embodiments, an ultrasonic transducer may be used to vaporize
the media in the vessel and thereby cause a mass of photosynthetic
microorganisms to collect around the transducer where they may
subsequently be harvested. Optionally, the vaporized media may be
collected and recirculated for use in the photobioreactor. The
microorganisms may then be harvested and/or dehydrated by any
methods known in the art.
[0119] The photobioreactor may further comprise a separating
apparatus for separating the removed portion of media containing
the photosynthetic microorganisms into a liquid phase and into a
solid phase (which contains the microorganisms). The separating
apparatus is preferably a filter but depending on the type of
microorganisms, other separating means known to one skilled in the
art may be used.
[0120] In an exemplary method, the photobioreactor disclosed herein
may be used to propagate one or more photosynthetic microorganisms
(e.g., a polyculture) such as algae that produce biomolecules such
as fatty acids, phycobiliproteins such as C-Phycocyanin,
allophycocyanin, phycoerythrin, biofuels such as phytol, and other
various petrol fuel substitutes. In one embodiment, two or more
algae may be propagated together that have natural environments
that are similar in salinity and dissimilar in temperature such as
algae selected from the group consisting of isochrysis aff.
galbana, pavlova lutheri, arthrospira platensis, chlorella
pyrenoidosa, synechococcus elongates, including naturally occurring
or genetically modified/recombinant strains of the foregoing.
[0121] Such a combination of algae provides the benefit that while
the temperature of the liquid media in the vessel may change, there
will typically be at least one alga that propagates at a low
temperature and at least one alga that propagates at a higher
temperature. Algae are then propagated in the photobioreactor as
provided herein and ultimately separated from the growth medium
after the algae reach a desired density. After separation of the
algae from the growth media, DHA is obtained from the algae and
optionally purified.
[0122] Without further description, it is believed that one of
ordinary skill in the art may, using the preceding description and
the following illustrative examples, make and utilize the agents of
the present disclosure and practice the claimed methods. The
following working examples are provided to facilitate the practice
of the present disclosure, and are not to be construed as limiting
in any way the remainder of the disclosure.
Examples
Example 1: Construction of Pebble
[0123] A pebble (also referred to herein as lamp assembly), for
practice of the methods provided herein, may be constructed by any
known methods and materials in the art and comprises a light source
and a barrier surrounding the light source.
[0124] A. Pebble Assembly
[0125] A pebble may be constructed from a circuit panel (e.g.,
pentagonal shaped panels) with pre-mounted LED array. A pebble
section (e.g., two or more affixed pentagon-shaped circuit boards)
is prepared by bonding each with liquid cyanoacrylate (1
three-panel section, 2 four-panel sections) along internal edges,
not connecting one section to the other. The cyanoacrylate is given
approximately 3 hours to dry. After drying, three sections are
press fit together into a dodecahedral shape, with one side of the
dodecahedron kept open for connectivity purposes (e.g., wire
access), and allowed to dry and settle into shape overnight. Next,
the three main sections are taken apart. On each of the two 4-sided
pebble sections, using lead-free solder only, ground connections
with 28 gauge wire are soldered to the central panel. This is
repeated with a 12-volt connection, making connection to same
central panel as ground and with 3-sided portion, using any of the
3 sides as the main connection panel. The ground connection from
the main panel on the 3-panel section is then connected to the main
panel on one of the 4-panel sections with 28 gauge wire and
lead-free solder. The second 4-panel section's main panel is
connected to the first 4-panel section on the panel on the far end
on the section adjacent to the main panel (sharing a side with the
main panel, with only one other side bonded to a panel). To this
last panel, now connecting all the ground on one circuit and all
the 12V connection on another, solder a 22-gauge wire to each
circuit (length to be determined by even pebble distribution
throughout barrel (approximately 10''-40'')). Next, the panels are
press fit together, taking care to carefully arrange wires within.
The pebbles are then filled with 10-12 glass beads for weight.
[0126] B. Pebble Case Assembly
[0127] A case for the pebble may be prepared using a polyethylene
(Nalgene) jar with dimensions to allow for the circumference of the
pebble. A 1/4'' hole is drilled in the center of the lid of the
jar. Next, a silicone gasket is inserted 1/4'' through the wall of
the lid. A male 1/4'' brass through-wall barbed fitting is then
screwed from underside of lid and attached to a 1/4'' silicone
gasket from top of the lid. A female 1/4'' brass fitting is then
screwed to the male fitting from the top of the lid. Next, 11/2''
of 1/4'' inner diameter flexible silicone tubing is attached to the
lid gasket using cyanoacrylate as a bonding agent. 1/4'' OD rigid
LLDPE white tubing (length to be determined by optimal pebble
dispersion within barrel) is then attached to connection tubing
using cyanoacrylate as a bonding agent. The base of a jar is then
filled with a number of clear glass beads (e.g., 14) for weight and
light dispersion. The assembled pebble is then placed within the
jar and seated upon glass beads. 22 gauge wire is then threaded
through the gasket and tubing. Subsequently, the jar is filled to
maximum with white mineral oil and the lid is tightly screwed on
expelling as much air as possible. Excess oil adhered to the
outside of the jar is removed and the jar is then rinsed with
reverse osmosis water.
[0128] C. Pebble Case Embodiments
[0129] FIGS. 12 to 35 are diagrams showing embodiments of lamp
assemblies. In each of the embodiments, the lamp assemblies 108
include circuit boards 113 connected together in a specific
geometry based on the dimensions of the casing. Each of the lamp
assemblies includes an inlet and an outlet to facilitate the flow
of immersion oil. The inlet and outlet are dimensioned to
mechanically connect to the connection lines 602. Further, the
inlet includes electrical connectors 111 (which are not shown).
[0130] For example, FIGS. 12 and 13 show a lamp assembly 108 that
includes six circuit boards 113 connected together to form a
double-pyramid. The lamp assembly 108 includes a casing 1202 that
is dimensioned to accommodate the six circuit boards. In
particular, the casing 1202 is bulb-shaped, which provides an
efficient propagation of light to a surrounding liquid culture
medium. The lamp assembly 108 receives immersion oil via the inlet
1204. The immersion oil exits the lamp assembly 108 at output 1206.
Further, electrical connectors 111 are routed through the inlet to
the circuit boards 113 (not shown). In some examples, the
electrical connectors 111 contact a first circuit board. Electrical
traces are used to electrically connect the other circuit boards to
the first circuit board. In other examples, each of the circuit
boards 113 is connected to the electrical connectors 111.
[0131] FIGS. 14 to 16 show schematic diagrams of the lamp assembly
108 of FIGS. 12 and 13. In particular, FIG. 14 shows a
top-perspective view of the lamp assembly and FIG. 16 shows an
enlarged view of a case connection 1504 of the casing 1502 shown in
FIG. 15. In this embodiment, the casing 1502 is formed as two
separate halves joined together at joint 1600. The casing 1502 is
formed as separate halves to enable the circuit boards to be
installed inside the casing 1502. As shown in FIGS. 15 and 16, the
joint 1600 may be mechanically sealed by the dimensioning of each
half of the casing 1502. Alternatively, the joint 1600 may be
chemically sealed using an (substantially transparent)
adhesive.
[0132] FIGS. 17 to 22 show another embodiment of a lamp assembly
108. In this embodiment, a casing 1702 is more spherical than the
casing 1502. In addition, FIGS. 19 to 22 shows that the casing 1702
is formed of two halves that are connected together using tabs
1902. In particular, FIGS. 21 and 22 show how the halves of the
casing 1702 are mechanically connected by each of the tabs
contacting a corresponding reception area.
[0133] FIGS. 23 to 28 show different embodiments of circuit boards
within the lamp assemblies. In particular, FIG. 23 shows a
top-perspective view of a lamp assembly 108 shown in FIG. 24. The
lamp assembly 108 includes a triangular matrix of circuit boards
113 connected together to form a cube. The connection of the
circuit boards 113 enables light to be transmitted in substantially
all directions for optimal culture growth.
[0134] FIG. 25 shows a diagram of an eight-sided cube formed from
triangular circuit boards 113. FIG. 26 shows a diagram of
three-sided cube formed from triangular circuit boards 113. FIG. 27
shows a diagram of six-sided double pyramid formed from triangular
circuit boards 113. FIG. 28 shows a diagram of six-sided cube
formed from triangular circuit boards 113. It should be appreciated
that other embodiments can include greater number of sides to form
a structure that is substantially spherical.
Example 2: Construction of Photobioreactor
[0135] A photobioreactor for practice of the methods provided
herein may be constructed by any known methods and materials in the
art and is constructed by assembly of a barrel, lid, and CO.sub.2
stone (also referred to herein as a lamp assembly).
[0136] A. Assembly of Barrel
[0137] In an exemplary method, using a standard 60 gallon
polypropylene barrel (open head), the barrel is prepared by
drilling 8 equidistant 1/2'' holes 41/2'' from the base of the
barrel. Equidistant between two adjacent holes, a 9th 1/2'' hole is
drilled 21/2'' from the base of the barrel. Next, using an exacto
blade, excess plastic is removed and the drill hole edges are
smoothed.
[0138] From the inside of the barrel, using a wrench, a 1/4''
barbed male nylon fitting is screwed through each of the drilled
holes. Next, from outside the barrel, 1/2'' silicone washer
fittings are then attached onto the barbed male fittings which are
then screwed onto 1/4'' nylon barbed female fittings and tightened.
13/4'' length of rigid (thick wall) 1/4'' tygon tubing is then
attached to the male fitting on the outside of the barrel.
Subsequently, 1/2'' length of black 1/8'' black silicone tubing is
slid over the barbed fitting of a large air stone. The air stone
assembly is then inserted into the tygon tubing and the process is
repeated for all 1/2'' fittings at the 41/2'' height. A 3'' length
of rigid tygon tubing (thick wall) is then attached to an external,
female barbed fitting. This process is then repeated for all 9
fittings. Next, 130'' of rigid white LLPDE tubing is attached to
all 8 fittings at the 41/2'' height. A push-to-connect adaptor
fitting.sup.i and push-to-connect ball valve are then attached to
the 9th (21/2'' height) fitting. Subsequently, 2'' of clear 1/4''
tygon tubing (thin wall) is attached to each of the 130'' white
rigid LLDPE tubes. 81/2'' lengths of 1/8'' black silicone tubing
are then connected to barbed fittings of an 8-way polypropylene
manifold. Each rigid LLPDE 130'' tube is connected to the 2'' long
clear tygon tubing connector (thin wall), which is then connect to
the manifold.
[0139] B. Assembly of Lid
[0140] In an exemplary method, seven holes are drilled into a black
polypropylene lid using a 1/2'' plastic drill bit, with a
configuration as shown in FIG. 11A.
[0141] Any excess plastic is then cleaned away from the drilled
edges with an exacto blade and smoothed. From the bottom of the
lid, 1/4'' male barbed nylon fittings are placed in the holes and
screwed together with 1/4'' female barbed nylon fittings from the
top side of the lid. Silicone washers may be placed between the
female fitting and the top side of the lid. Distribution of lights
using other fittings to be determined by the strain(s) intended for
cultivation. Next, 2'' length of rigid tygon tubing (thick wall) is
attached to each male nylon 1/4'' fitting in the lid. A length of
rigid LLPDE tubing is then attached to each connection, except the
central location (length to be determined by even distribution of
pebbles throughout barrels) as shown in FIG. 11B.
[0142] Subsequently, 2'' rigid tygon connector tubing is then
connected to an end of the rigid LLPDE tubing. The pebble wiring is
run through the rigid LLPDE tubing and out through the barbed
fittings in the top of the lid. Connector tubing is then attached
to the base of pebble's brass fitting using liquid cyanoacrylate as
a binding agent. This process is repeated as needed for desired
pebble distribution. For protection from salt contamination,
flexible latex tubing is attached to top, female end of lid
fittings and around all wires as needed based on distribution of
saltwater spray.
[0143] C. Assembly of CO.sub.2 Stone
[0144] In an exemplary method, 30'' length of rigid white LLDPE
tubing is attached to a central fitting with connector tube. Next,
the dome apex is marked on the female hemisphere of a 2 part 100 mm
clear acrylic sphere. The corners of an equidistant triangle, with
sides approximately 2'' in length, are then marked out on the male
hemisphere and centered on the dome apex. A 1/4'' rotary tool
grinding bit is then used to make preliminary guide holes for each
marking. Next, a 1/2'' rubberized grinding bit is used to make
final holes centered on guide holes. Excess plastic from drilling
is removed from the holes is removed with clippers and an exacto
blade until the holes are smooth. A 1/2'' rubber through-wall
gasket is then attached to each of the holes. Next, a 1/2'' brass
through-wall barbed fitting is attached to the central hole in the
female hemisphere with a gasket. The male hemisphere is then filled
with 450 grams of clear glass beads. Subsequently, cyanoacrylic
bonding agent is applied to the bottom male hemisphere and the male
and female hemisphere are press fit together. The cyanoacrylic
bonding agent is allowed to dry for approximately one hour. Next,
2'' length of rigid tygon is attached to a brass fitting barb using
cyanoacrylic bonding agent. Again, the cyanoacrylic bonding agent
is allowed to dry for approximately one hour. The assembled sphere
and connector tube are then attached to rigid white tubing attached
to central lid fitting.
Example 3: Use of Photobioreactor for Growth of Photosynthetic
Microorganism
[0145] A photobioreactor provided herein may be used in methods for
growing one or more photosynthetic microorganisms. Such methods may
employ a four step process including: 1.) determining optimal
environmental conditions (OEC); 2.) staging and inoculation of
production environment; 3.) growing a photosynthetic microorganism
to an extractable mass; and 4.) selective extracting of mature
cells.
[0146] A. Determination of Optimal Environmental Conditions
(OEC)
[0147] When starting up a new or unknown strain it is necessary to
determine optimal values for all growth variables including:
salinity, nutrient concentrations, EM frequency (RWB ratio), EM
cycle, and rate of airflow. OEC may be determined by using an
approximation of natural environmental conditions (NEC) as a
starting point. Beginning with three Generation 4 reactor chambers
(2 liter capacity) for each variable to be tested, process of
elimination is used to narrow down the options. For example,
chamber 1 comprises an experimental variable at a concentration 60%
greater than its NEC, chamber 2 comprises the experimental variable
at its NEC, and chamber 3 comprises the experimental variable at a
concentration 60% less than its NEC. Speed of growth of the strain
is determined by observation of the rate in change of Diffused
Optical Density (DOD). If/then for DOD:
[0148] If C1>C2>C3, Then Round 2 baseline=C1 with variances
of +/-15%;
[0149] If C2>C1&C3, Then Round 2 baseline=C2 with variances
of +/-15%;
[0150] If C3>C2>C1, Then Round 2 baseline=C3 with variances
of +/-15%
[0151] This process is continued for 4 rounds and repeated for each
experimental variable to determine the new strain's OEC values.
[0152] B. Staging and Inoculation of Production Environment
[0153] Using the OEC values determined above, a Generation 5
Reactor (30 liter capacity) is prepared to those levels determined
in A above. Salinity levels are set, pumps are activated, and
nutrients are added to the reactor in accordance with such
predetermined levels. The nutrients are given approximately two
hours to mix without light. After the nutrients have mixed, the
Reactor is seeded with at least 10% live culture. Based on
continuous testing of water nutrient levels, the culture is fed as
needed for the next 3-10 days based on the growth rate of the
strain. When DOD has reached a level where individual LED's are no
longer visible, a generation 7 reactor is prepared using the same
method as described above with 90% of the culture in Generation 5
Reactor as inoculation for Generation 7 Reactor.
[0154] C. Growth to Extractable Mass
[0155] Once growth is established in the Generation 7 Reactor, the
following conditions should be continually monitored: pH, ammonia,
nitrate, nitrite, phosphate, dissolved CO.sub.2, dissolved O.sub.2,
system air flow, system pressure, and diffused optical density. As
these conditions deviate from OEC in response to microorganism
growth steps must be taken to maintain OEC across these parameters.
For example, if a Gen 7 cyanobacterial culture has tripled in
density over a 48 hour period (observable through increase in DOD
and Turbidity) and Ammonia has decreased to 0 ppm then a diluted
addition of concentrated fertilizer should be added to return
Ammonia to OEC. Additionally, for example, if a Gen 7 culture has
been run continuously for 3 months and in response to multiple
feedings and a buildup of digestive waste pH has increased off OEC
to 8.4 then a diluted addition of organic acid (i.e. citric acid)
should be added to return pH to that culture's OEC for pH.
[0156] As exponential growth of photosynthetic microorganisms
continues, DOD will continue to increase in proportion with culture
density. When culture density has doubled three times from the
point of its 10% inoculation it is at an ideal point to provide
seed inoculation to other reactors. At a 10% level of inoculation,
the culture can be used to seed 9 other reactors of comparable size
with enough mass left over to self-inoculate its own restart. At a
given point culture density will reach a level where the reactor
primary fluid can no longer hold the culture in suspension. This
point is identified by the increased levels of
accumulation/agglomeration in low flow points along the reactor
bottom and sides in the presence of OEC. The point at which the
system tips toward this condition will be referred to as Peak
Suspended Mass (PSM). When PSM is achieved the culture must be put
through selective extraction, comprehensive extraction or used to
seed other reactors if growth rates are to be maintained. Time to
PSM and culture density at PSM vary widely depending on strain and
OEM.
[0157] D. Selective Extraction of Mature Cells
[0158] Prior to the point when PSM is achieved, one of two
extraction configurations must be established: 1.) Gen 7 Outlet
Valve=>Pump=>Filter=>Gen 7 Return; or 2.) Gen 7 Outlet
Valve=>Filter=>Pump=>Gen 7 Return. The configuration is
determined based on the characteristics of the strain being
filtered. The effectiveness of selective filtration is based on the
size variance between mature and immature cells in the given
strain(s) and proper selection of the pore diameter of the
filtration medium. Filtration pore diameter should be greater than
the diameter of the immature cells and less than the diameter of
the mature cells by a preferred margin of >25%. Once one of the
above configurations is set and PSM is achieved the following steps
can be taken: a.) Gen 7 outlet valve moved from CLOSED to OPEN; b.)
once pump is primed, pump moved from OFF to ON; c.) once filter bag
is full and air pressure has equalized across filter bag, filter
cap can be moved from CLOSED to OPEN; d.) optional: if the filter
has an installed outlet valve on its return line that valve should
be moved from CLOSED to OPEN before the pump and filter are moved
to their engaged configuration. At no point should filter pressure
exceed 15 psi, 10 psi for smaller diameter bags (<5 microns).
Filtration should be monitored closely with new strains as
filtration time varies widely with strain.
[0159] Unless otherwise indicated, all numbers expressing
quantities of ingredients, properties such as molecular weight,
reaction conditions, and so forth used in the specification and
claims are to be understood as being modified in all instances by
the term "about." Accordingly, unless indicated to the contrary,
the numerical parameters set forth in the specification and
attached claims are approximations that may vary depending upon the
desired properties sought to be obtained by the present disclosure.
At the very least, and not as an attempt to limit the application
of the doctrine of equivalents to the scope of the claims, each
numerical parameter should at least be construed in light of the
number of reported significant digits and by applying ordinary
rounding techniques.
[0160] Notwithstanding that the numerical ranges and parameters
setting forth the broad scope of the disclosure are approximations,
the numerical values set forth in the specific examples are
reported as precisely as possible. Any numerical value, however,
inherently contains certain errors necessarily resulting from the
standard deviation found in their respective testing
measurements.
[0161] The terms "a," "an," "the" and similar referents used in the
context of describing the disclosure (especially in the context of
the following claims) are to be construed to cover both the
singular and the plural, unless otherwise indicated herein or
clearly contradicted by context. Recitation of ranges of values
herein is merely intended to serve as a shorthand method of
referring individually to each separate value falling within the
range. Unless otherwise indicated herein, each individual value is
incorporated into the specification as if it were individually
recited herein. All methods described herein can be performed in
any suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g., "such as") provided herein is intended
merely to better illuminate the disclosure and does not pose a
limitation on the scope of the disclosure otherwise claimed. No
language in the specification should be construed as indicating any
non-claimed element essential to the practice of the
disclosure.
[0162] Groupings of alternative elements or embodiments of the
disclosure disclosed herein are not to be construed as limitations.
Each group member can be referred to and claimed individually or in
any combination with other members of the group or other elements
found herein. It is anticipated that one or more members of a group
can be included in, or deleted from, a group for reasons of
convenience and/or patentability. When any such inclusion or
deletion occurs, the specification is deemed to contain the group
as modified thus fulfilling the written description of all Markush
groups used in the appended claims.
[0163] Certain embodiments of this disclosure are described herein,
including the best mode known to the inventors for carrying out the
disclosure. Of course, variations on these described embodiments
will become apparent to those of ordinary skill in the art upon
reading the foregoing description. The inventor expects skilled
artisans to employ such variations as appropriate, and the
inventors intend for the disclosure to be practiced otherwise than
specifically described herein. Accordingly, this disclosure
includes all modifications and equivalents of the subject matter
recited in the claims appended hereto as permitted by applicable
law. Moreover, any combination of the above-described elements in
all possible variations thereof is encompassed by the disclosure
unless otherwise indicated herein or otherwise clearly contradicted
by context.
[0164] Specific embodiments disclosed herein can be further limited
in the claims using, consisting of, or and consisting essentially,
of language. When used in the claims, whether as filed or added per
amendment, the transition term "consisting of" excludes any
element, step, or ingredient not specified in the claims. The
transition term "consisting essentially of" limits the scope of a
claim to the specified materials or steps and those that do not
materially affect the basic and novel characteristic(s).
Embodiments of the disclosure so claimed are inherently or
expressly described and enabled herein.
[0165] It is to be understood that the embodiments of the
disclosure disclosed herein are illustrative of the principles of
the present disclosure. Other modifications that can be employed
are within the scope of the disclosure. Thus, by way of example,
but not of limitation, alternative configurations of the present
disclosure can be utilized in accordance with the teachings herein.
Accordingly, the present disclosure is not limited to that
precisely as shown and described.
[0166] While the present disclosure has been described and
illustrated herein by references to various specific materials,
procedures and examples, it is understood that the disclosure is
not restricted to the particular combinations of materials and
procedures selected for that purpose. Numerous variations of such
details can be implied as will be appreciated by those skilled in
the art. It is intended that the specification and examples be
considered as exemplary, only, with the true scope and spirit of
the disclosure being indicated by the following claims. All
references, patents, and patent applications referred to in this
application are herein incorporated by reference in their
entirety.
* * * * *