Maximization of algae lipid yield Scenedesmus dimorphus for the production of biodiesel

Polythecnic University of Puerto Rico
Chemical Engineering Department
Course – CHE- 5916
Capstone Project Presentation
Maximization of ScenedesmusDimorphusLipid Yield for the Production of Biodiesel
Group:
Sara Currás Medina
Gustavo Mendez Santos
Carlos A. Ramos Encarnación
Germano Salazar Benites
Advisor:
Dr. Alessandro Anzalone
Date:
July 24, 2009

Acknowledgment
Prof. Sylvia M. VélezVillamil
Biology Department
University of Puerto Rico at Humacao
Prof. Edgardo González, Ph.D.
Bureau of Forest Services, Director
Department of Natural and Environmental Resources
Alessandro Anzalone, Ph.D. – Advisor
Chemical Engineering Department, Director

Agenda

Problem Statement

Introduction

Introduction: Algae Fuel
Introduction

ResearchDescription

ResearchObjectives

Research Contributions
Determine if the use of CO2 during algae cultivation is beneficial to its growth and lipid content.
Maximize the lipid content varying the urea concentration and nutrient depravation time.
Detailed documentation of the process.

Literature Review

Biodiesel fromAlgae
Production of Oil from differentcrops
Advantages:
The yields of oil and fuels are much higher than competing energy crops.
Grows practically anywhere, ensuring no competition with food crops.
Excellent bioremediation agents - they have the potential to absorb massive amounts of CO2 and can play an important role in sewage and wastewater treatment.
Only feedstock that has the potential to completely replace world's consumption of transportation fuels.

Microalgae vs. Macroalgae
Microalgae
Macroalgae
Unicellular
Microscopic (µm)
Saline or fresh water
Grow extremely quickly
Large amounts of lipids within their cell structure
Multicellular
Macroscopic (up to 60 m)
Saline or fresh water
Grow extremely quickly
Food, medicine, fertilizer

Properties of Green Microalgae

Microalgae Strain

Scenedesmusdimorphus

Production of Lipids from Microalgae
Light (Photons)
CO2
O2
Microalgae
(Photosynthetic CO2 fixation)
Nutrients
(N, P, Si)
Biomass (carbon)
Lipid storage
Carbohydrate storage

Cultivation

Cell growth

Temperature

CO2

Nutrients
After Carbon, Nitrogen is the most important nutrient.
Contributes to the biomass production.
Influence on algae lipid yield
Response to nitrogen limitations, mechanism of survival, increase lipid content.
Stops its divisions and start to store energy in the form of lipids.

Nitrogen; Urea
Urea
Each strain prefers different source.
Kansas State University
Best source for Scenedesmusdimorphusgrowth.
Replacing nitrogen source KNO3

Harvesting

Flocculation

Extraction

Lipid Extraction
Concentrated Algae
Cell Disruption
Mechanical
Chemical
Press
Solvents
Filtering
Distillation
Algae Oil

Lipid Extraction - Solvents

Transesterification

Transesterification Reaction

Proposed Experiment
Co2 vs. non Co2
Algae Culture
Daily
Cell Count
N Measure
After 17 days
Biomass Wt.
Lipid Wt.

Cell Count

Nitrogen Measure

Biomass and Lipid Weight

Proposed Experiment
Optimal Lipid Yield
Algae Culture
1.2, 1.8, 2.4 g/L Urea
Daily
Cell Count
N Measure
After N Consumption
Biomass Wt.
Lipid Wt.

Cell Count

Nitrogen Measure

Proposed Experiment
Biodiesel Production
Algae Culture
Daily
Cell Count
After a fixed period
Process the oil
Obtain Biodiesel

Stock Culture
Prepare the Stock Culture (SC) in a beaker of 1000mL.
1.
algae
1000mL
SC

Stock Culture
Leave the algae to grow and reproduce for about 7 days before using it.
12:12
2.
Agitation
SC
90°F

Growth Media
Prepare the Growth Media(GM) in an Erlenmeyer of 1000mL.
1.
GM

Cultivation
To prepare one culture for any experiment.
1000mL
SC
GM
227mL GM
23mL SC
250mL
Culture

CO2 vs. Without CO2
For this experiment we want to determine if the use of carbon dioxide during algae cultivation is beneficial to its growth and lipid content.
90°F
12:12
250mL
250mL
Culture
Culture
x3
x3

Measuring Algae Growth
Measure the growing rate using a hemacytometer every day.
100X
magnification
250mL
Culture

Measuring Nitrogen Concentration
Spectrophotometer
250mL
Culture

Lipid Yield
After certain days of algae culturing, extract 100 mL
from the culture and transfer it to a 250 mL beaker.
Using aluminum sulfate flocculate the sample.
With a 1.5µm filter , separate the sample using a
vacuum Erlenmeyer of 1000 ml.
Measure and record the weight of a 500 mL bottle
with cap.
Deposit the biomass contained in the filter in to the
weighted bottle and measure again.

Lipid Yield
For every gram of algae biomass add 18 ml of
Hexane/Isopropyl (3:2) (solvent) and agitate manually until
the biomass is dissolved.
Measure the weight of a vacuum Erlenmeyer of 1000 mL.
With a 1.5µm filter, separatethe sample using the previously
weighed Erlenmeyer.
Put the Erlenmeyer with the filtrated solution in the hood.
Using a heat plate, apply heat to the solution (95˚C) to
evaporate the solvent solution.
With the solution dry, measure the weight and evaluate the
results.

Dry Weight Biomass
After 17 days put a filter paper in an oven at 75˚C for
5 hours.
Measure the weight of the paper.
Take 30 mL from the culture and filtrate in a vacuum
Erlenmeyer of 1000mL.
After filtrating the sample put the filter paper in the
oven at 75˚C for 5 hours.
Leave the sample overnight in the desecator.
Measure the weight of the sample.
Compare the weight of the dry lipids vs. the dry
biomass.

Optimization of Lipid Yield
Prepare 15 cultures in 15 beakers of 250 mL with three different concentrations (1.2, 1.8 and 2.4 g/L Urea)
1.2 g/L Urea
1.8 g/L Urea
2.4 g/L Urea
Measure the growing rate
and the nitrogen
consumption every day.

Biodiesel
Measure the growing rate every day.
1.
Measure the nitrogen consumption every day.
2.
90°F
When the culture is prepared to be processes, flocculate the sample
3.
12:12
With a 1.5µm filter , filtrate the sample using a vacuum.
4.
For every gram of biomass add 18 ml of a mixture of Hexane/Isopropyl (3:2)
5.
After agitation, filtrate the sample again.
6.
Separate by distillation the solvent from the oil.
7.
CO2 or Air

Transesterification
1. Add the extracted oil to a 1 liter flask.
2. In another flask mix KOH with ethanol
3. Heat the ethanol to dissolve KOH if needed.

Transesterification
4. Mix KOH and ethanol blend into algae oil
and agitate vigorously.
5. After 120 minutes of reaction time, allow time
for separation. The mixture will separate into
two layers biodiesel on top, glycerin on bottom.
Biodiesel
Glycerin
6. Separate the biodiesel on another flask.

Transesterification
8. Allow 24-48 hours for water to settle,
biodiesel will float to the top and become
clearer.
7. Place the biodiesel in a glass column
and spray water into the top..
Biodiesel
Water
9. Separate the biodiesel from water.

References
“Oil Crisis”. Health and Energy. 13 July 2009 <http://healthandenergy.com/oil_crisis.htm>.
Sheehan, John, Terri Dunahay, John Benemann, and Paul Roessler. A Look Back at the U.S. Department of Energy’s Aquatic Species Program—Biodiesel from Algae. Colorado: NationalRenewableEnergyLaboratory, 1998.
“Scenedesmus Dimorphus-Algae Culture”. Algae Depot. 2009. 12 June 2009 <http://www.algaedepot.com/servlet/the-1/Scenedesmus-dimorphus--dsh--Algae/Detail>.
“Kunikane. S, M. Kakeko, and R. Maehara. Growth and Nutrient Uptake of Green Alga, Scenedesmus Dimorphous, Under a Wide Range of Nitrogen/Phosphorous Ratio-I. Setsunan, Japan: University of Setsunan, 1984.
Shen, Ying, Zhijian Pei, Wenqiao Yuan, and Enrong Mao. Effect of Nitrogen and Extraction Method on Algae Lipid Yield. Kansas State: University of Kansas State, 2009.
Tzann, Stelios T. “Non Mechanical Methods”. Tutorial on Cell Disruption. 3 June 1996. 7 July 2009 <http://128.113.2.9/dept/chem-eng/Biotech Environ/DOWNSTREAM/disrupt.htm>.

Thanks for
listening!

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Maximization of algae lipid yield Scenedesmus dimorphus for the production of biodiesel

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Maximization of algae lipid yield Scenedesmus dimorphus for the production of biodiesel Presentation Transcript