By: Kaitlyn Summerfield -...
Transcript of By: Kaitlyn Summerfield -...
![Page 1: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/1.jpg)
By: Kaitlyn Summerfield
![Page 2: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/2.jpg)
Bioresource extraction and why is it important
Project design
Cell disruption methods
Implications of results
Possible future developments
![Page 3: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/3.jpg)
sustainable technology
Investigates natural avenues of resource capture
Reduces our dependence on finite resources
Implications for many industries
Qualitative analysis of cell disruption methods on three distinct samples of algae
![Page 4: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/4.jpg)
Chlorella sp. (ACW-1)- locally discovered species
Landfill polyculture, dominant species: Kirchneriella sp. And Scenedesmus sp.
Euglena sp. found at Crone’s Cradle Conserve
Autoclaving
Freeze-fracture
Surfactant (TWEEN 80)
Cell grinding
Microwaving
Solvents : Octanol, Butanol, Isopropanol
![Page 5: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/5.jpg)
A B
CSample Species controls: (A)- ACW-1 500x magnification, (B)- Euglena sp. 500 x magnification, (c)- LFS Polyculture 500x magnification
![Page 6: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/6.jpg)
Creation of ice crystals with cellular structure during freezing, thawing process causes cellular expansion catalyzes rupture
4 separate trials of 15 minute freeze/15 minute thaw periods
![Page 7: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/7.jpg)
B
C
BA
Phase 4 of Freeze- Thaw Fracture Study:(A)- ACW-1 500x magnification, (B)-Euglena sp. 500 x magnification, (C)-LFS Polyculture 500x magnification
![Page 8: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/8.jpg)
“Surfactant”- surface active agent, main ingredient in soaps, hydrophilic and hydrophobic components
Works by increasing the solubility of the cellular membrane
![Page 9: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/9.jpg)
A B
CTWEEN 80 testing- (A)- ACW-1 500x magnification, (B)- Euglena sp. 500x magnification, (C)- LFS Polyculture 500x magnification
![Page 10: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/10.jpg)
•Preferred method for cellular disturbance in plant tissues
• Tests conducted with and without surfactant
![Page 11: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/11.jpg)
Cell grinding: (A)- ACW-1 500x magnification, (B)- Euglena sp. 500x magnification,(C)-LFS Polyculture 500x magnification
A B
C
![Page 12: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/12.jpg)
•Novel method for investigating the effects of heat on cellular disruption
•Tests conducted with surfactant ( total time 6 minutes 40 seconds) and without (total time 2 minutes)
• Highly energy intensive- would not be economically viable on a larger scale
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A B
CMicrowaving with Surfactant:(A)- ACW-1 500x magnification, (B)- Euglena sp. 500x magnification, (C)- LFS Polyculture500x magnification
![Page 14: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/14.jpg)
•Method of testing the effects of high heat and high pressure on cellular disruption
•30 minutes, 120˚ c, 120 psi
•Another energy intensive disruption technique
![Page 15: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/15.jpg)
Autoclave 30 min 120˚ C, 120 psi: (A)- ACW-1 500x magnification, (B)- Euglena sp. 500x magnification, (C)- LFS Polyculture 500x magnification
AB
C
![Page 16: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/16.jpg)
Disintegration of cellular membrane
visible analysis of pigment extraction and microscopic observation of cellular disturbance
Octanol- most promising and most environmentally friendly of the solvents tested
![Page 17: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/17.jpg)
A B
CEuglena sp. solvent results: (A)-Euglena after octanol addition 500x magnification, (B)- Euglena after butanol addition 500x magnification, (C)- Euglena after isopropanol addition 500x magnification
![Page 18: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/18.jpg)
Euglena sp. sample after pigment extraction with octanol
Concentrated cell pellet
Water component from sample
Octanol and carotene pigment
![Page 19: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/19.jpg)
•Euglena responsive to every cell disruption test
•Cell wall compositions responsible for results differentiation
•Use of Euglena sp. could increase the economic viability of bioresource extraction
![Page 20: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/20.jpg)
• Commercially valuable resources:-Astaxanthin- antioxidant and pigment enhancer used in aqua culture and food industry-paramylon- cosmetic industry
• Octanol- pigment extraction recycling loop- reuse after extraction
• Possible exploration of other routes of environmentally friendly pigment extraction- vegetable oils
![Page 21: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/21.jpg)
Questions?
![Page 22: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/22.jpg)
•Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press vol.3 Physiology
•Green, J. (1963). The Occurrence of Astaxanthin in the Euglenoid TrachelomonasVolvocina. Comp. Biochem. Physiol. (9) p. 313-316.
•Harrison, Susan T.L. (1991). Bacterial Cell Disruption: A Key Unit Operation in the Recovery of Intracellular Products. Biotech. Adv. (9) p. 217-240.
•John, D.M., Whitton, B.A., and Brook, A.J. (2008). The Freshwater Algal Flora of the British Isles: An Identification Guide to Freshwater and Terrestrial Algae. Cambridge: Cambridge University Press.
•Kang, Chang Duk and Sim, Sang Jun. (2007). Selective Extraction of Free Astaxanthinfrom Haematococcus Culture Using a Tandem Organic Solvent System. Biotechnol. Prog. (23) p.866-871.
![Page 23: By: Kaitlyn Summerfield - Biogasbiogas.ifas.ufl.edu/Internships/2010/files/kaitlynsummerfield.pdf · •Buetow, Dennis E. ( 1982). The Biology of Euglena. New York: Academic Press](https://reader031.fdocuments.us/reader031/viewer/2022011819/5e9f6e78ae2c342c1b2050fd/html5/thumbnails/23.jpg)
•Kang, Chang Duk and Sim, Sang Jun. (2008). Direct Extraction of Astaxanthinfrom Haematococcus Culture Using Vegetable Oils. Biotechnol Lett. (30) p. 441-444.
•Marten, Gerald G. (1987). The Potential of Mosquito-Indigestible Phytoplankton for Mosquito Control. Journal of the American Mosquito Control Association 3(1): 105-106.
•Mendes-Pinto, M.M., Raposo, M.F.J., Bowen, J., Young, A.J., and Morais, R. (2001). Evaluation of Different Cell Disruption Processes on Encysted Cells of Haematococcus pluvialis: Effects on Astaxanthin Recovery and Implications for Bio-availability. Journal of Applied Phycology 13: 19-24.
•Nakano, Yoshihisa, Urade, Yoshihiro, Urade, Reiko, and Kitaoka, Shozaburo. (1987). Isolation, Purification, and Characterization of the Pellicle of Euglena gracilis z. Journal of Biochemistry 102: 1053-1063.