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Evaluating techniques of quantifying lipid yield and cell density of Chlorella protothecoides (Kruger) grown on waste glycerol

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Title: Evaluating techniques of quantifying lipid yield and cell density of Chlorella protothecoides (Kruger) grown on waste glycerol
Authors: Collins, Topaz Palakika
Advisor: Adolf, Jason E.
Keywords: Alternative energy
Microbiology
Conservation biology
biofuel
BODIPY 505/515
show 4 moreChlorella protothecoides
glycerol
heterotrophic
NucRed Live 647

show less
Issue Date: Dec 2015
Abstract: Chlorella protothecoides (Krüger) is ubiquitous as a research organism for biofuels. The genus consists of species that are photosynthetic, heterotrophic, and mixotrophic, and cellular lipid yield can be relatively high compared to other algal species. The research presented here addressed the differences between conventional and novel methods of quantifying lipid yield and cell density of C. protothecoides produced on various glycerol media. Conventional methods of light microscopy, gravimetric lipid analysis, and colony forming units, were compared with BODIPY 505/515 lipid-stained cells and NucRed Live 647 nucleic acid stained cells analyzed by flow cytometer. This is the first report of using a dual stain technique to simultaneously quantify cell density and lipid yield of an algal species for biofuel production. Results indicated that there is no significant difference (p > 0.05) between BODIPY 505/515 and NucRed Live 647 for cell density. There is no significance between the dyes and hemocytometer counts (p>0.05). Lipid analysis indicated no significant difference between flow cytometer lipid per cell and %oil/vol (p>0.05), especially for UTEX B25. This indicates that the novel method of utilizing BODIPY 505/515 or NucRed Live 647, singularly or in concert, is comparable to conventional methods of lipid and cell density analyses. Because all three strains of C. protothecoides obtained cell densities of between 107 and 109 cells/mL and highest oil yield was about 45% per cell, waste glycerol has the potential to become a viable feedstock for Hawaii’s growing biofuel industry.
Pages/Duration: 43 pages
URI/DOI: http://hdl.handle.net/10790/2594
Appears in Collections:Tropical Conservation Biology and Environmental Science



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