developing new working protocols and bioreactor designs to enhance biomass growth and energy yield...
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Developing New Developing New Working Protocols Working Protocols
and Bioreactor and Bioreactor Designs to Enhance Designs to Enhance Biomass Growth and Biomass Growth and
Energy Yield in Energy Yield in Schizochytrium Schizochytrium limacinumlimacinum and and C.reinhardtiiC.reinhardtiiJeffrey Yau and Christina Jeffrey Yau and Christina
GeorgeGeorge
Manhasset Science ResearchManhasset Science Research
BackgroundBackground The use of The use of
bioreactors to bioreactors to combat the growing combat the growing problem of problem of greenhouse gases greenhouse gases has been extensively has been extensively studied in recent studied in recent decades.decades.
(Chisti, 2007)(Chisti, 2007)http://www.ieagreen.org.uk/newsletter/dec80/images/biofixation.JPG
•The U.S. has reported a 3.3% increase of carbon dioxide emissions in the past year•Mostly due to car emissions and industrial factories. (Hopwood,2007)
http://photos.mongabay.com/08/0423methaneglobal.jpg
The red line shows the trend together with seasonal variations. The black line indicates the trend that emerges when the seasonal cycle has been removed.
Graph 1
Raceway Pond DesignRaceway Pond Design
A raceway pond is made of a closed loop recirculation channel
(Chisti, 2007)
http://www.agric.wa.gov.au/content/SUST/BIOFUEL/110407_Biodieselfrommicroalgae.pdf
Original Design Modified Design
Taken: January 17, 2009
Tubular Photo Tubular Photo Bioreactor DesignBioreactor Design
Consists of straight Consists of straight transparent tubes either made transparent tubes either made out of glass or plastic (also out of glass or plastic (also known as solar collectors)known as solar collectors)
(Chisti, 2007)(Chisti, 2007)
http://www.agric.wa.gov.au/content/SUST/BIOFUEL/110407_Biodieselfrommicroalgae.pdf
Original Design
Taken: January 17, 2009
Modified Design
Control: Airlift DesignControl: Airlift Design
A self-A self-contained contained bioreactorbioreactor
Utilizes a Utilizes a baffle to re-baffle to re-circulate the circulate the bacteria in bacteria in suspensionsuspension
Taken on: December 21, 2008
Organism: S.limacinumOrganism: S.limacinum Contains pigments Contains pigments
for photosynthesisfor photosynthesis Known to contain Known to contain
EPA, DHA, and EPA, DHA, and omega-3 fatty omega-3 fatty acidsacids
Reliable source of Reliable source of oil production for oil production for biodieselsbiodiesels
(Kamlangdee, 2003)(Kamlangdee, 2003) http://roweb.cityu.edu.hk/researchreport/2002-2003/Project/020.jpg
Organism: C.reinhardtiiOrganism: C.reinhardtii
Contains an Contains an enzyme called enzyme called hydrogenase that hydrogenase that allows creation of allows creation of hydrogen hydrogen
(Tiede, 2008)(Tiede, 2008) Ability to Ability to
produce produce hydrogen under hydrogen under anoxic conditionsanoxic conditions
(Fouchard, 2005)(Fouchard, 2005)http://en.wikipedia.org/wiki/Chlamydomonas_reinhardtii
Magnified 3000X
Tubular Photo Bioreactor Tubular Photo Bioreactor design for algal culturesdesign for algal cultures
Molina Grima; et al (2001): Experiment on Molina Grima; et al (2001): Experiment on tubular photobioreactors using tubular photobioreactors using P. P. tricornutumtricornutum
Tested tubular diameter on the amount of Tested tubular diameter on the amount of sunlight that penetrated through the sunlight that penetrated through the culture brothculture broth
Polyunsaturated fatty acids Polyunsaturated fatty acids production by production by
Schizochytrium sp.Schizochytrium sp. Kamlangdee Kamlangdee
(2003): Experiment (2003): Experiment on polyunsaturated on polyunsaturated fatty acids fatty acids production by production by Schizochytrium sp.Schizochytrium sp.
Found single isolate Found single isolate reliable in reliable in production of DHAproduction of DHA
Hydrogen as Clean Fuel Via Hydrogen as Clean Fuel Via Continuous Fermentation by Continuous Fermentation by
Anaerobic Photosynthetic Anaerobic Photosynthetic Bacteria, Rhodospirillum Bacteria, Rhodospirillum
rubrumrubrum Observed the Observed the effect of light effect of light intensity, intensity, agitation, and agitation, and liquid dilution liquid dilution rate on hydrogen rate on hydrogen productionproduction
Use of biocatalyst Use of biocatalyst can be considered can be considered alternative to alternative to Fischer Tropsch Fischer Tropsch synthetic synthetic reactionsreactions
(Najafpour, 2003)(Najafpour, 2003)http://www.vurup.sk/pc/vol45_2003/issue3-4/pdf/14.pdf
PurposePurpose
Therefore the purpose of this Therefore the purpose of this experiment was to create a experiment was to create a bioreactor design that would bioreactor design that would enhance growth rate and energy enhance growth rate and energy yield in yield in Schizochytrium limacinumSchizochytrium limacinum and and C.reinhardtiiC.reinhardtii
HypothesisHypothesis
Null Hypothesis:Null Hypothesis: No significant difference No significant difference will be found in the growth of will be found in the growth of C.reinhardtii and S.limacinum in either C.reinhardtii and S.limacinum in either bioreactor.bioreactor.
Alternate Hypothesis:Alternate Hypothesis: The growth of The growth of C.reinhardtii and S.limacinum will be C.reinhardtii and S.limacinum will be greatest in the Tubular Photobioreactor greatest in the Tubular Photobioreactor when exposed to carbon dioxide, with when exposed to carbon dioxide, with respect to pH levels.respect to pH levels.
MethodologyMethodologyProblem: What is the most efficient design for a photo bioreactor to enhance the energy yield and growth rate of Schizochytrium limacinum and C.reinhardtii?
Control Group:-Growth medium under normal conditions.-Growth rate in Airlift Bioreactor
Independent Variable 1:Growth of Schizochytrium limacinum
Dependent Variables:-Oil Extracted from Hexane-Hydrogen collected from C.reinhardtii-Three different sized tubes (0.01m, 0.012m, 0.019m)
An ANOVA test will be used to statistically analyze the data (p<.05). The Scheffe post hoc test will be used.
Schizochytrium limacinum will be obtained from atcc.org. The Glucose Yeast Extract Medium will contain 1g of Yeast Extract, 1g of Peptone, 5.0g of Glucose, and 1L of 15% Natural Seawater in a 1000mL Volumetric flask. The yeast extract, glucose, and the Natural Seawater will be obtained from Carolina Biological. Peptone will be obtained from Flinn Scientific.Growth Projection: 4 weeks
Independent Variable 2:Growth of C.reinhardtii
Photo Bioreactor
Measurement of Biomass yield (once daily from start of experimentation) using Aquafluor Fluorometer
C.reinhardtii will be obtained from Carolina, the Biological Vendor. The C.reinahrdtii will be cultured in a fresh water tank, with a 12 hour supply of light, and oxygen. The algae will be kept at 70 degrees Fahrenheit and cultured with a 10% Algal growth medium.
Raceway pond
Measurement of Biomass yield using Spectrophotometer (Wavelength at 610nm)
Measurement of the effect of carbon dioxide on pH levels (Using Co2 Sensor on GLX and pH paper)
Graph 1: Comparison of the growth of C.reinhardtii, between the Airlift, Tubular and raceway pond Photo Bioreactors for Trial 1. There was a significant increase in growth in the Tubular Photo Bioreactor as compared to the Airlift and Raceway Pond. (p=.019)
Growth of C.reinhardtii in Airlift Compared to Tubular PhotobioreactorGrowth of C.reinhardtii in Airlift Compared to Tubular Photobioreactor and Raceway Pond (25% Medium)and Raceway Pond (25% Medium)
Time (Day)
Tra
nsm
itta
nce
70
75
80
85
90
95
100
1/8/
09
1/9/
09
1/10
/09
1/11
/09
1/12
/09
1/13
/09
1/14
/09
1/15
/09
1/16
/09
1/17
/09
1/18
/09
1/19
/09
1/20
/09
1/21
/09
1/22
/09
1/23
/09
Airlift
Tubular
Raceway
N=1N=144
Graph 2: Comparison of the growth of C.reinhardtii between the Raceway Pond and Tubular Photo Bioreactor. There was a significant increase in growth in the Tubular Photobiroeactor as compared to the Raceway. (p=.019)
Growth of C.reinhardtii in Tubular Photobioreactor vs. Raceway PondGrowth of C.reinhardtii in Tubular Photobioreactor vs. Raceway Pond
Time
Tra
nsm
itta
nce
60
65
70
75
80
85
90
95
100
1/8/
09
1/9/
09
1/10
/09
1/11
/09
1/12
/09
1/13
/09
1/14
/09
1/15
/09
1/16
/09
1/17
/09
1/18
/09
1/19
/09
1/20
/09
1/21
/09
1/22
/09
1/23
/09
Tubular
Raceway
N=1N=144
Graph 3: Comparison of the growth of C.reinhardtii between the Tubular and the Raceway Pond after CO2 exposure. No significant difference was found.
Growth of C.reinhardtii after COGrowth of C.reinhardtii after CO22 Exposure Between Tubular Exposure Between TubularPhotobioreactor and Raceway PondPhotobioreactor and Raceway Pond
Time (Day)
Tra
nsm
itta
nce
70
75
80
85
90
95
100
1/15/09 1/16/09 1/17/09 1/18/09 1/19/09 1/20/09 1/21/09 1/22/09 1/23/09
Tubular
Raceway
N=N=77
Graph 4: Comparison of the growth of C.reinhardtii between the Airlift, Raceway, and Tubular Photo Bioreactor after carbon dioxide exposure for Trial 1. There was no significant difference in growth in all bioreactors.
Growth of C.reinhardtii after COGrowth of C.reinhardtii after CO22 Exposure in Airlift Compared to Tubular Exposure in Airlift Compared to Tubular Photobioreactor and Raceway PondPhotobioreactor and Raceway Pond
Time (Day)
Tra
nsm
itta
nce
60
65
70
75
80
85
90
95
100
1/15/09 1/16/09 1/17/09 1/18/09 1/19/09 1/20/09 1/21/09 1/22/09 1/23/09
Airlift
Tubular
Raceway
N=7N=7
Graph 5: Shows the percent change in absorbance of C.reinhardtii in all three bioreactors for Trial 1. The Tubular Photobioreactor (red) grew C.reinhardtii with the most percent change in absorbance out of all three bioreactors while the Raceway Pond grew the organism with the least percent change in absorbance.
4.9
18
2.7
0
5
10
15
20
25
30
Airlift Tubular Raceway
Percent Change in Absorbance of C.reinhardtii in Bioreactors for Trial 1
Bioreactors
Per
cen
t (%
)
Graph 6: Shows the percent change in absorbance of C.reinhardtii in all three bioreactors for Trial 2. The Raceway Pond (light blue) grew C.reinhardtii with the most percent change in absorbance out of all three bioreactors while the Airlift grew the organism with the least percent change in absorbance.
Percent Change in Absorbance of C.reinhardtii in Bioreactors for Trial 2
1.5
3.9
9.7
0
2
4
6
8
10
12
Airlift Tubular Raceway
Bioreactors
Per
cen
t (%
)
Graph 7: Comparison of the growth of C.reinhardtii, between the Airlift, Tubular and Raceway Pond photo Bioreactors for Trial 2. There is no significant difference between the bioreactors. However, comparing the transmittance numbers (1/29-2/5) to the carbon dioxide levels before exposure, there is a significance. (p=.015)
Growth of C.reinhardtii in Bioreactors (10% Medium)
Time (Day)
Tra
nsm
itta
nce
80
82
84
86
88
90
92
94
96
98
100
1/26/09 1/27/09 1/28/09 1/29/09 1/30/09 1/31/09 2/1/09 2/2/09 2/3/09 2/4/09 2/5/09
Airlift
Tubular
Raceway
N=N=99
Graph 8: Comparison of the growth of C.reinhardtii between the Airlift, Raceway, and Tubular Photo Bioreactor after carbon dioxide exposure for Trial 2. No significance was found when compared to carbon dioxide levels measured.
Growth of C.reinhardtii After CO2 Exposure in Bioreactors (Trial 2)
Time (Day)
Tra
ns
mit
tan
ce
80
82
84
86
88
90
92
94
96
98
100
1/29/09 1/30/09 1/31/09 2/1/09 2/2/09 2/3/09 2/4/09 2/5/09
Airlift
Tubular
Raceway
N=N=66
DiscussionDiscussion A Tubular Photobioreactor is a suitable A Tubular Photobioreactor is a suitable
environment for growth of algaeenvironment for growth of algae Performance of the Tubular Performance of the Tubular
Photobioreactor surpassed the control Photobioreactor surpassed the control bioreactorbioreactor
Daily exposure to carbon dioxide did not Daily exposure to carbon dioxide did not greatly effect pH levels in bioreactorsgreatly effect pH levels in bioreactors
Growth in Tubular Bioreactor was greater Growth in Tubular Bioreactor was greater than growth in the Raceway Pond than growth in the Raceway Pond possibly due to more efficient pumppossibly due to more efficient pump
ConclusionConclusion Data supports the Alternate HypothesisData supports the Alternate Hypothesis The Tubular Photo Bioreactor The Tubular Photo Bioreactor
demonstrated a greater amount of demonstrated a greater amount of growth as compared to the Raceway growth as compared to the Raceway PondPond
There was no significance regarding There was no significance regarding the Carbon Dioxide levels when the Carbon Dioxide levels when pumped through the Tubular, Raceway pumped through the Tubular, Raceway Pond and Airlift BioreactorsPond and Airlift Bioreactors
LimitationsLimitations
Errors while using the Spectrophotometer Errors while using the Spectrophotometer occurred, causing incongruous data: Possibly occurred, causing incongruous data: Possibly caused by cuvette or contaiminationcaused by cuvette or contaimination
Possible errors in GLX Xplorer readings for Possible errors in GLX Xplorer readings for carbon dioxidecarbon dioxide
Possible bacterial contamination in bioreactorsPossible bacterial contamination in bioreactors
Future StudiesFuture Studies
Revision of Tubular Photobioreactor and Raceway Pond designs
Testing various tube diameters Using grown C.reinhardtii and
S.limacinum from bioreactors to extract hydrogen and oils, respectively, to test energy content
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