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Microbial biodiesel production
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- 2. Introduction to biodiesel History Rudolfdiesel was the inverter of biodiesel, estimated nearly 100 years ago. It was developed in the year 1890s.
- 3. What is Biodiesel? •Alternative fuel for diesel engines • Madefrom vegetable oil, animal fat, microbes • Lower emissions, High flash point (>300°F), Safer • Biodegradable, Essentially non-toxic. Fatty Acid Alcohol Glycerin Vegetable Oil BiodieselFA FAFA FA
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- 5. Characteristics Of Biodiesel Liquidvaryingin color Immiscible inwater Highboiling point of 360–640°F (182–338°C) Low vapor pressure: < 2 mmHg Specific gravity between0.86 & 0.90 Vapor density > 1 Less hazardous in terms of flammability
- 6. Biodiesel As LubricantAnd Solvent Biodiesel can be used straightas a machinery lubricant. Biodiesel’s solvent properties may be used to clean dirty or greasy engine or other machine parts.
- 7. Advantages of Biodiesel Renewableenergy source Less polluting Utilizes excess production of soybeans for manufacture Can distribute through existing diesel fuel pumps Can use in existing oil heating systems and diesel engines Can be mixed with petroleum diesel at any concentration and time
- 8. Disadvantages Of Biodiesel Moreexpensive Could harm rubber hoses in engines Requires energy to: Produce biodiesel from soy crops & sow, fertilize and harvest Requires frequent filter changing Requires improvement in distribution infrastructure
- 9. Relative Greenhouse GasEmissions 020 40 60 80 100 120 140 160 Gasoline CNG LPG Diesel Ethanol 85% B20 Diesel Hybrid Electric B100 B100 = 100% Biodiesel B20 = 20% BD + 80% PD
- 10. Relative emissions: Dieseland Biodiesel 0 20 40 60 80 100 120 Total Unburned HCs CO Particulate Matter **NOx Sulfates PAHs n-PAHs Mutagenicity CO2 Percent B100 ** B20 Diesel
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- 12. Trans esterification CH2OOR1 catalystCH2OH | | CHOOR2 + 3CH3OH 3CH3OORx + CHOH | | CH2OOR3 CH2OH Triglyceride 3 Methanols Biodiesel Glycerin R1,R2,andR3arefattyacid alkylgroups.
- 13. Biodiesel production byalkaliprocess Enzymatic BiodieselProduction Trans esterification
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- 15. Biodiesel Reaction Vegetable Oilor Animal Fat (100 lbs.) + Methanol or Ethanol (10 lbs.) Biodiesel (100 lbs.) + Glycerin (10 lbs.) In the presence of a catalyst Combining Yields
- 16. NRRaje Feb 06 Conventionalfeed stocks Rapeseed,themajorsource(>80%) Sunfloweroil (10%, ItalyandSouthernFrance) Soybeanoil (USA) Palmoil (Malaysia) Linseed,oliveoils (Spain) Cottonseedoil (Greece) Beef tallow(Ireland), lard,Jatropha(Nicaragua),)
- 17. Microorganisms available forbiodieselproduction Microalgae Bacteria Fungi Yeast Biodiesel production using microbial lipids, which is named as single cell oils (SCO), has attracted great attention in the whole world. Lipidsfromall cannotbeconverted into biodiesel mainly dueto less yield Oleaginous microorganisms are able to accumulate lipids above the 20% of their biomass, on dry basis.
- 19. Stagesof Lipid accumulation ... Cellsreach limit of obesity → stop accumulating. Lipid accumulation → cells expand Cells convert C → storage lipid (intracellular) Exhaustion of N, cells stop divide Microorganisms grow and multiply until a certain time Prepare medium: high Carbon, low Nitrogen
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- 21. • Can growrapidly •Live in harsh conditions due to their unicellular orsimple multicellular structure. Of the 33,000 known species of algae at least forty are considered oleaginous Goodcandidatesfor biodieselproduction, higher photosynthetic efficiency higher biomass production and faster growth compared to other energy crops MICROALGAE
- 22. continue.. • Can beinducedto accumulate substantial quantities of lipids thus contributing to a high oil yield. • Someofthe common algae chlorella, crypthecodinium, cylindrotheca, dunaliella, isochrysis, nannochloris, nannochloropsis, neochloris, nitzschia, phaeodactylum, porphyridium, schizochytrium, tetraselmis, etc,…. average lipid content- (1-70%) maximum- 90%
- 23. How to obtainbiodiesel from algae? Pickingupthebestalgae Growingthealgae Extraction Transesterification Biodiesel
- 24. Choosing an Algae Important characteristicsof Algae High % oftotal biomass is oil Maintains a high % of oil even under stress Compatible with ourregional climate
- 25. Where To GrowIt Extensionsonto our water treatmentplants Clean up our waste and generate fuel Agriculture runoff Water ponds Algae could well grow on salty water aswell assoft water
- 26. How Is AlgaeGrown? Opensystem TubularPBR Flat plate PBR Closed systems(photo bioreactor)
- 27. Extraction Solvent extraction Mechanical extraction Supercriticalfluid extraction
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- 31. DIFFERENCE B/W BIODIESELFROM ALGAE AND OTHER PLANT/VEGETABLE OILS • The yield • According to some estimation, theyield (per acre) of oilfrom algaeis over 200 times theyield from the best-performingplant/vegetable oils.
- 32. ADVANTAGES OF MICROALGAEAS A SOURCE OF BIODIESEL • High Yield -low cost ofproduction • Algae can grow –In places away from farm land (Nodestruction tofood chain) –Sewages –Near to power plants (takes CO2from smokestacks and yields oil) • Oil Productivity –Greaterthan bestproducing oil crops • Higher grade protein→Animal Feed • Balanced N: P ratio→Organic Fertilizers
- 33. Biodiesel from Fungus Oleaginousfungi hasalso been considered as potential oilsources for biodiesel production because they accumulate large amounts of lipids Among these microorganisms, particular attention has beendedicated to various oleaginous zygomycetes species, such as Mortierella isabelina and Cunninghamella echinulata, which may accumulate up to 86% and 57% of lipids in the dry biomass, respectively
- 34. Biodiesel from Fungus FungalBio Mass Mucorcircinelloides Three Different Solvent Systems: Chloroform:Methanol (C:M), Chloroform:Methanol:Water (C:M:W) n-hexane Acid Catalyst (BF3, H2SO4 and HCl ) Materials
- 35. Biodiesel from FungalBio Mass Extraction oflipids Solvent +Dried biomass Mixture Ultrasonication Centrifugation RotaryEvaporation Lipids
- 36. Biodiesel from FungalBio Mass
- 38. Rhodosporidium sp., Rhodotorula sp.and Lipomycess species can accumulateintracellular lipids as high as 70% of their biomass dry weight. •Cryptococcus curvatus oleaginous yeast Accumulate storage lipid up to >60% ona dry weight basis) Biodiesel from yeasts
- 39. Biodiesel from yeasts Media Freezedrying Lipidextraction Tran esterification Biodiesel
- 40. Media Pre-culture YM medium wasusedaspre-culturetocultivateoleaginousyeastcells o glucose as a carbon source o Peptone o yeast extract Sterlisation - 120ºC temperaturefor20minutes Semi-synthetic medium was usedas mainculturetogrowoleaginousyeasts o Glucose, xyloseor a mixture of both were used as carbon sources o nitrogen sources (NH4Cl) o phosphate buffer (KH2PO4) o Mineral-elementsolution (CaCl2×2H2O ,FeSO4×7H2O ,citric acid×H2O) MediumpH - 5.8 Main culture
- 41. Freeze drying o Collectedsamples were centrifuged at 10 min andwashed with sterile water once, then centrifuged again o Thepellets weretransferred into pre-weighed vials and kept at -50°C until freezedrying
- 42. Lipid extraction Chloroform- MethanolMethod AddingChoroform: Methanol mixture Centrifugation Freezedried pellets + HCL and incbated at 55° C Solvent Evaporation Extracted lipids
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- 44. Biodiesel from Bacteria Bacteriacan accumulateoilof about 20-40% of dry biomass Arthrobactersp.-40% Acinetobactercalcoaceticus-38% Have a superiority in the production of biodiesel dueto Highest growthrate(reachhuge biomassonlyneed 12–24h) Easyculturemethod. Actinomycetegroup high amountoffattyacids (up to70%ofthe cellular dryweight) using glucose undergrowth-restrictedcondition
- 45. Downside • Veryfewareoil producer •Only a fewbacteria accumulate complicated lipoid. • It is difficult toextract becausethese lipoid aregenerated in the outer membrane • So there is no industrial significance in the actual production of biodiesel by using oleaginous bacteria as raw material
- 46. Quality ofbiodiesel frommicrobes • Biodiesel fuel, in the form of FAME, is nowmanufactured in many countries. • Relevant standard to assessbiodiesel are; ASTMD6751( In USA) EN14214(In EU, intended for vehicle use) EN14213(In EU, for use as heatingoil)
- 48. Conclusion •At presentplant oilis the main feedstock for biodiesel production. • However, at present biodiesel is not competitive with conventional fuels in the whole world duetohigh cost of production. • Production of microbial based diesel can be an economical beneficial. • However, it still needs lots of improvement which could be done using the various biotechnological techniques and methods. • Developing high lipid content microorganisms for biodiesel production from microbes are promising option in future and opens a possibility for academic research
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