1. WAJIHA SEERAT Ph.D. Scholar Department of Botany

2. 

3.   Introduction  Biofuel Feedstocks: Potential Contributions of Synthetic Biology  Synthetic Biology and Biodiesel Production  Indirect Synthesis of Biodiesel from Microbial Oils  Direct Synthesis of Biodiesel Using Cell Factories  Biosynthesis of Ethanol  Biosynthesis of Fatty Acid Ethyl Esters (Biodiesel)  Conclusions and Future Perspectives Contents

4.  Global Warming Non renewable fuel resources Two major Problems

5.  Entensive use of Fossil fuels Green house gas emissions Instability of oil supplies Fluctuations in prices Environment damage

6.   Biofuel is energy made from living matter, usually plants -- renewable energies, emit less than fossil fuels, and have received increasing attention in the transition to low- carbon economy  Bioethanol, biodiesel are types of biofuels  Bioethanol is an alcohol produced from corn, sorghum, potatoes, wheat, sugar cane, even biomass such as cornstalks and vegetable waste  Biodiesel is made from plant oils such as soybean oil, palm oil and rapeseed oil. INTRODUCTION

7.   White biotechnology centers on the bioproduction of fuels and chemicals from renewable sources  About 90% of the current biofuel market is represented by biodiesel and bioethanol  Bioethanol is not seen as an ideal biofuel for the future  Low energy density  Incompatibility with the existing fuel infrastructure INTRODUCTION

8.  Biodiesel is preferable to petrodiesel  Environmental friendliness  Renewability  Reduced emissions  Higher combustion  Efficiency  Improved lubricity  Higher levels of safety Cont’d…

9.   Chemically, biodiesel ------ mixture of fatty acid alkyl esters (FAAEs).  Commonly used method ------ in vitro Transesterification process Cont’d…

10. 

11. 

12.   Issues related to large-scale commercial viability of biodiesel production  High cost  Limited availability of vegetable oils  In vitro transesterification reaction presents some unresolved issues  need to use large amounts of toxic compounds (sodium hydroxide, sulfuric acid, or methanol)  high cost of isolation and immobilization of enzyme catalysts Cont’d…

13.   Increasing interest in developing microbial processes  for the production of biodiesel from a wide range of other raw materials  Technologies that use living cells to synthesize products  biodegradable  require less energy  create less waste during production APPROACHES

14.   Lignocellulosic agricultural residues  edible and nonedible crops ---- for the production of biofuels  More than 95% of global biodiesel production now begins from edible vegetable oils  account for about 80% of the total production cost  Use of nonedible vegetable oils is significant for biodiesel production in developing countries,  tremendous demand for edible oils as food. Biofuel Feedstocks: Potential Contributions of Synthetic Biology

15. 

16.   Increasing attention to the use of microbial oils as biodiesel feedstock  produced by certain oleaginous microorganisms  Lignocellulosic biomass --- largest known renewable source of carbohydrates  Synthetic biodiesel can be produced from this renewable carbon source Cont’d…

17.   Techniques used in conversion of lignocellulose into biofuels appears simple in theory  not fully established  The yield and productivity of biofuels synthesized in this way are not sufficient to meet current energy demands. Cont’d…

18.   In ethanol production,  Starches (maize, wheat, barley, etc.)  Sugar-rich biomass (grasses, maize leaves, beets, sugar cane, etc.)  Feedstocks most commonly used for their bioconversion  Ethanol is one of the substrates  used for in vivo synthesis of biodiesel Cont’d…

19.   Synthesis of biodiesel using microbes --- alternative to conventional technologies  Microbial biodiesel production from two different angles:  By indirect synthesis from microbial oils  By direct biodiesel synthesis using redesigned cell factories Synthetic Biology and Biodiesel Production

20.   Oils derived from oleaginous microbes  represent promising raw materials for biodiesel production  through transesterification using the plant-based process  Microalgae are photoautotrophic microorganisms that can convert CO2 directly to lipids,  which can then be used for biofuel production, particularly for biodiesel Indirect Synthesis of Biodiesel from Microbial Oils

21. 

22.   Tools from synthetic biology used to convert certain autotrophic microalgae into heterotrophic microorganisms  Introduction of non-natural metabolic pathways into the autotrophic microalgae  allowing cultivation using an organic carbon source instead of photosynthesis from sunlight Cont’d…

23.   Ethanol produced endogenously --- for in vivo synthesis of fatty acid ethyl esters (FAEEs) with exogenously added FFAs.  Microbial FFAs can be used as feedstock for in vivo production of biodiesel Direct Synthesis of Biodiesel Using Cell Factories

24.  Biosynthesis of Ethanol

25.   Direct synthesis of biodiesel from microbes produces primarily FAEEs, which show better performance as a fuel than FAMEs  An important discovery related to this was a novel bifunctional wax ester synthase/acyl-CoA: diacylglycerol acyltransferase (WS/DGAT) isolated from Acinetobacter baylyi strain ADP1.  This enzyme can synthesize wax esters from alcohols and fatty acids Biosynthesis of Fatty Acid Ethyl Esters (Biodiesel)

26. 

27.   An E. coli strain was manipulated  for improvement of the biosynthesis of FFAs  for derivatives including fatty esters Cont’d…

28.   Metabolic engineering and synthetic biology allowing new technologies to be developed  As evident in the design and refinement of microbial cell factories amenable to industrial applications  Biodiesel production through the use of microbial systems has marked a turning point in the field of biofuels  The possibility of developing a synthetic host for efficient target-molecule production presents great opportunities for further biofuel research CONCLUSION & FUTURE PERSPECTIVES

29.   Investment of significant amounts of time and effort is still required in order to produce a better host  Carrying novel metabolic pathways to lead to satisfactory biofuel production  More engineering of all of the genes needed to produce the desired pathway is not sufficient to confer a novel characteristic to a recombinant cell, since there are always many other unexplored pathways as well. Cont’d…

30.   Veronica L. C., A. Rodriguez and H. A. Cristobal. 2011. The Role of Synthetic Biology in the Design of Microbial Cell Factories for Biofuel Production Journal of Biomedicine and Biotechnology, 0.1155/2011/601834, 1-9. REFERENCE

31. 

32. 