From Plants to Energy Efficient Lignin Degradation BIOFUELS

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From Plants to Energy: Efficient Lignin Degradation BIOFUELS Margarette Francis, Maham Hijazi, Reeda Mahmood, From Plants to Energy: Efficient Lignin Degradation BIOFUELS Margarette Francis, Maham Hijazi, Reeda Mahmood, Riza Unabia & Manaum Zain 1

FOSSIL FUELS: USAGE AND CONCERNS v“Fossil Fuels” in today’s world v Green house gasses, FOSSIL FUELS: USAGE AND CONCERNS v“Fossil Fuels” in today’s world v Green house gasses, climate change and global warming v Why still use them? v Economically, it is more beneficial v Hybrid electric car v inefficent 2

ALTERNATIVE OPTIONS… v. Biofuels Ethanol v Increasingly common option in agricultural and forestry industry ALTERNATIVE OPTIONS… v. Biofuels Ethanol v Increasingly common option in agricultural and forestry industry in Canada v DISADVANTAGE presence of lignin v Genomics to overcome this problem v An exemplary model SOYBEAN 3

WHY SOYBEAN? v. Soybean (Glycine max) v. Abundant all over the world v. Grown WHY SOYBEAN? v. Soybean (Glycine max) v. Abundant all over the world v. Grown in Southern and Eastern Ontario v. Breeding advances (E. O) v. Significant source of protein and oil 4

ETHANOL AS AN ENERGY SOURCE… v. Reduces greenhouse gases v. Does not contribute to ETHANOL AS AN ENERGY SOURCE… v. Reduces greenhouse gases v. Does not contribute to carbon dioxide emissions (34% reduction) v. Input of energy required to produce ethanol has dropped by 36% v. Gives more energy compared to other renewable sources 5

WHY IS LIGNIN A BARRIER? v Barrier for cellulose and hemicellulose extraction v Lignin WHY IS LIGNIN A BARRIER? v Barrier for cellulose and hemicellulose extraction v Lignin removal improve the digestibility of the biomass v Mechanical and chemical treatments are used to breakdown lignin v rate of reaction is slow v Altering the pathway in order to reduce lignin is more effective v rate of reaction is comparatively faster Wood Composition. (2007). Retrieved from http: //learn. forestbioenergy. net/learning-modules/module-6/unit-1/lesson-1 Yarris, L. (2013, November 13). Lignin-Feasting Microbe Holds Promise for Biofuels. States News Service. Retrieved from http: //www. highbeam. com/doc/1 G 1 -348985171. html? 6

THE EFFECT OF LIGNIN REDUCTION ON THE STRUCTURE OF PLANTS v Removing too much THE EFFECT OF LIGNIN REDUCTION ON THE STRUCTURE OF PLANTS v Removing too much lignin from a plant affects its phenotype v Research conducted on the plant Arabidopsis thaliana v Reduction of 64% of lignin, compensated by the increase in matrix polysaccharides or cellulose v Afingen technique was used for lignin reduction, producing functionally and structurally normal plants Stems & Vascular Tissue. (2012). Retrieved from http: //www. sciencepartners. info/? page_id=534 7

The Phenylpropanoid Pathway Reinprecht, Y. , Yadegari, Z. , Perry, G. , Siddiqua, M. The Phenylpropanoid Pathway Reinprecht, Y. , Yadegari, Z. , Perry, G. , Siddiqua, M. , Wright, L. , Mcclean, P. , & Pauls, K. (2013). In silico comparison of genomic regions containing genes coding for enzymes and transcription factors for the phenylpropanoid pathway in Phaseolus vulgaris L. and Glycine max L. Merr. Frontiers in Plant Science. Retrieved March 19, 2015, from http: //journal. frontiersin. org/article/10. 3389/fpls. 2013. 00317/full 8

A CLOSER LOOK…. Cinnamyl alcohol dehydrogenase (CAD) catalyzes the reaction that is involved in A CLOSER LOOK…. Cinnamyl alcohol dehydrogenase (CAD) catalyzes the reaction that is involved in the formation of the monolignols that make up lignin. CAD P-Hydoxyphenol lignin CAD Guaiacyl lignin CAD Syringyl lignin Baldoni, A. , Von Pinho, E. V. R. , Fernands, J. S. , Abreu, V. M. , & Carvalho, M. L. M. (2013). Gene Expression in the lignin biosynthesis pathway during soybean seed development. Genet. Mol. Res 12 (3) 2618 -2624 9

METHODS FOR GENOME EDITING v. Finger nucleases v. RNA interference v. Transcription activator-like effector METHODS FOR GENOME EDITING v. Finger nucleases v. RNA interference v. Transcription activator-like effector nucleases v Expensive and time-consuming v. A recent and more innovative process. . 10

CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS (CRISPRS) AND CRISPR-ASSOCIATED (CAS) PROTEINS CRISPR/CAS 9 SYSTEM CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS (CRISPRS) AND CRISPR-ASSOCIATED (CAS) PROTEINS CRISPR/CAS 9 SYSTEM 11

WHAT IS IT? vderived from a natural process found in bacteria to protect themselves WHAT IS IT? vderived from a natural process found in bacteria to protect themselves from pathogens vtargets genes for editing and regulating vcomparable to Photoshop . Horizon Licenses Harvard University Gene-Editing Technology. (2013). Drug Discovery & Development 12

TO INITIATE GENE MODIFICATION, sg. RNA (single guide RNA) Cas 9 nuclease Cas 9 TO INITIATE GENE MODIFICATION, sg. RNA (single guide RNA) Cas 9 nuclease Cas 9 complex 13

Protospacer Adjacent Motif (PAM) Target Sequence Gene of Interest 14 Protospacer Adjacent Motif (PAM) Target Sequence Gene of Interest 14

Non-Homologous End Joining (NHEJ) DNA repair pathway Stop Codon 15 Non-Homologous End Joining (NHEJ) DNA repair pathway Stop Codon 15

SUMMATION: CRISPR/CAS SYSTEM sg. RNA/Cas 9 complex binds to gene Cas breaks gene (double SUMMATION: CRISPR/CAS SYSTEM sg. RNA/Cas 9 complex binds to gene Cas breaks gene (double strand breaks) Induced mutation in gene sequence altered gene sequence dysfunctional CAD gene 16

WHAT MAKES A GOOD BIOFUEL? 1. Greenhouse Gas Reduction Potential 2. Cost-Effectiveness 3. Energy WHAT MAKES A GOOD BIOFUEL? 1. Greenhouse Gas Reduction Potential 2. Cost-Effectiveness 3. Energy Allocations for Crops 17

ABUNDANCE IN CANADA - ONTARIO v Soybean is one of the most widely used ABUNDANCE IN CANADA - ONTARIO v Soybean is one of the most widely used feedstock for biofuel production. Crop Production Data in Metric Tonnes 2012 2013 2014 Barley 165, 500 134, 600 151, 300 Corn 8, 598, 300 9, 007, 300 7, 600, 000 Soybean v Canada’s second leading crop. Crops 3, 401, 900 3, 238, 600 3, 791, 100 Canola 61, 200 49, 900 31, 300 Wheat, winter 1, 750, 000 2, 277, 900 1, 627, 500 Statistics Canada. CANSIM table 001 -0010 18

ENERGY ALLOCATIONS WCBA. Canada. http: //westerncanadabiodiesel. org/biodiesel/ Soybean has a much higher net energy ENERGY ALLOCATIONS WCBA. Canada. http: //westerncanadabiodiesel. org/biodiesel/ Soybean has a much higher net energy balance as compared to various other feedstocks 19

GREENHOUSE GAS EMISSIONS According to the Canadian Natural Resources model, GHGenius, soybean biodiesel and GREENHOUSE GAS EMISSIONS According to the Canadian Natural Resources model, GHGenius, soybean biodiesel and corn ethanol were found to reduce GHG emissions by 50% and 21%, respectively. According to the Greenhouse gases, regulated Emissions and Energy in Transportation (GREET) model for life cycle analysis, all soybean-derived fuels achieve a significant reduction (52– 107%) in fossil energy use and in petroleum use (more than 85%) 20

WHY IS IT GOOD FOR CANADA? Cost-effective Eco-friendly Sustainable Renewable 21 WHY IS IT GOOD FOR CANADA? Cost-effective Eco-friendly Sustainable Renewable 21




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