The biodiesel production from microbial oil of Rhodosporidiumtoruloides DEBB 5533 using sugarcane juice

1. Course Title: Biofuels and Biorefinery
Presented By: Huroom Aslam
Submitted To: Dr Zainab

2. Pilot scale biodiesel production from microbial oil of
Rhodosporidium
toruloides DEBB 5533 using sugarcane juice:
Performance in dieand preliminary economic study

3. INTRODUCTION
 Biodiesel is a fuel derived from renewable biomass.
 Main alternative to fossil fuel and received attention from all over the
world.
 Biodiesel produced from vegetable oils, animal fats, waste cooking
oils, lipids of yeast and microalgae.
 Chemically, biodiesel can be defined as:
“Fuel alkyl esters of long-chain carboxylic acids, produced from the
transesterification and/or esterification of vegetable, animal or
microbial fats and greases” 3

4. CONT…
 Biofuel does not contribute to atmospheric CO2 emissions.
 It is highly biodegradable therefore, has minimal toxicity.
 Global biodiesel production increased 13% to 29.4 billion liters in
2014.
 The top-producing countries were
United States (60 billion liters)
Brazil (29.9 billion liters)
Germany (4.3 billion liters)
China (3.9 billion liters)
Argentina (3.6 billion liters).
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5. CONT…
 Brazil is the lead producer of sugarcane (Saccharum spp.) in world.
 It is expected that 658 million tons will be harvested in 2015 and 2016.
 Its products and sub-products are widely used in the production of:
Sugar
Alcohol
Heat
Electricity
Biodiesel and bioplastic 5

6. CONT…
 Sugarcane juice, popularly known as ‘‘garapa,” contains:
15% (w/w) of fermentable sugars, an excellent substrate for microbial oil
production.
 Biodiesel produced by yeast is
Sustainable
More efficient and similar production costs than biodiesel from oilseeds
Strains grow quickly, and produce larger quantities of neutral lipid
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7. CONT…
 This work presents
A promising alternative for biodiesel production
Technology of SCO, with the non-GMO yeast Rhodosporidium toruloides
Grown in low-cost sugarcane media
 Experimental procedures included
i. Biomass production process development and scale-up
ii. Cell separation and disruption for intracellular oil extraction and
purification
iii. Microbial oil conversion to biodiesel
iv. Biodiesel purification and its performance in diesel engine tests.
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8. MATERIALS AND METHODS
Flowchart
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9. I. Strain For Microbial Oil Synthesis
 Rhodosporidium toruloides DEBB 5533 strain is a (non-GMO).
Inoculum medium 
Incubated at 32 ⁰C at 180 rpm for 12 h
Serial inoculum propagations at
10 mL, 150 mL, and 1000 mL.
Sugarcane-based medium (40 g/L)
Nitrogen source (urea)
pH was adjusted to 5.0
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10. II. Rhodosporidium toruloides DEBB 5533 Biomass
Production
 Oil production was initially conducted in 5L , 25L and 500L volumes
Fermentation carried out under fed-batch operational conditions
After 24 h
Bioreactor was fed with a carbon source (sugarcane juice)
After 48 h
Fermentation biomass settled down to the bottom until the concentration
reached 120 g/L.
10

11. III. Cell Disruption And Microbial Oil Recovery
 Microbial oil recovery
carried out
 Cell disruption by a chemical hydrolysis method using mineral acids
50 ⁰C under agitation of 60 rpm
 (After cell lysis) Medium with cells (the lipoproteic fraction)
sent to
 Separation unit 11

12. IV. Biooil Oil Extraction
Lower phase
recovered
Micelles
sent
Solvent evaporation
unit
Upper phase
(extracted
fraction) 20% of
incorporated micelles
separated by
Centrifugation
Centrifuged mass
(contained some
amount of oil )
re-extracted with
solvent
▪ Biooil extraction took place in three stages:
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13. V. Separation Of Micelles
Reboiler
2 ml sample withdrawn (every 5 min)
3 samples show same concentration of oil
recovered
Solvent
subsequent extraction
Oil reserved 13

14. VI. Biooil Analyses
 Analyses of microbial oil were performed via:
Peroxide value (PV) in the oil
Iodine Index (II) in the oil
Free fatty acids (FFA) in the oil
Saponification number (SN)
Gas chromatography
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15. VII. Biooil Conversion To Biodiesel
 Conversion of microbial oil to biodiesel involved
i. Acid Esterification (first stage)
Here conversion of all triglycerides into free fatty acids takes place
ii. Transesterification (second stage)
iii. Methanol and sodium hydroxide were added to the reactor
98%
fatty acid
esters
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16. VIII. Recovery And Analysis Of Esters
 Esters were purified by 2 operations:
i. An extraction with water washing
ii. Addition of selective adsorbent material
Sedimentation started
supernatant centrifuged purified esters at the top
adsorbent retained in bottom
Esters then analysed by standards
16

17. IX. Engine Tests
 Test of produced biodiesel as well as different fuels carried out in an
Agrale/M95W engine for:
Emissions
Consumption
Performance
 Two different mixtures of biodiesel and diesel used for the tests:
B5 (5% biodiesel in mineral diesel)
B20 (20% biodiesel in petroleum diesel)
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18. X. Economic Analysis
 Based on
Costs of the medium
Energy involved in each step of the
Biooil and biodiesel production
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19. RESULTS AND DISCUSSION
I. Microbial Oil Production And Recovery
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20. 20

21. II. Biooil Recovery And Analysis
 Efficiency of 99% of oil recovery was attained
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22. III. Analysis Of The Microbial Biodiesel
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23. IV. Microbial Oil Biodiesel Engine Tests
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24. 24

25. 25

26. V. Economic Analysis Of The Microbial Biodiesel
Process Production
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27. CONCLUSIONS
 Microbial oil from yeast Rhodosporidium toruloides DEBB 5533 attained:
High biomass
Biooil at low-cost medium based on sugarcane juice
 Engine tests showed
Great performance
Lower pollutant emissions compared to biodiesel from soybean oil.
 This process used grass (sugarcane) as raw material not a food-feed
product so estimated price of biodiesel is low. 27

28. 28

29. 29
Reference
“Pilot scale biodiesel production from microbial oil of Rhodosporidium
toruloides DEBB 5533 using sugarcane juice: Performance in diesel
engine and preliminary economic study”
Carlos Ricardo Soccol , Carlos José Dalmas Neto , Vanete Thomaz Soccol , Eduardo
Bittencourt Sydney , Eduardo Scopel Ferreira da Costa , Adriane Bianchi Pedroni
Medeiros , Luciana Porto de Souza Vandenberghe.