Succinic Acid- Presentation

1. Microbial Synthesis of Succinic Acid from Typha Grass Hydrolysate and Its Application in Biopolymer Synthesis and as Co-plasticizer By Yakindra P Timilsena (111332) Examination Committee Prof. Athapol Noomhorm Prof. Sudip K Rakshit Dr. Anil Kumar Anal

2. Main Idea Renewable/Green Chemicals Cheap and Under-utilized resources Biodegradable polymer with better properties

3. Introduction Succinic Acid is a C4 dicarboxylic acid with molecular formula C 4 H 6 O 4 , molecular weight 118.09 and melting point 185－190 C . It is predicted to be one of the most widely used platform chemicals which can be produced from renewable feedstocks ( Bechthold et al. 2008). Broad range of industrial applications -as a source of food, pharmaceuticals, surfactants, detergents, antifoam agents, in the production of resins, polymers, paints, cosmetics and inks ( Isar et al. 2006). Biobased succinic acid can be a suitable alternative to its petrochemical equivalent.

4. Introduction Glycerol is a by-product of biodiesel industries and can be used as a good plasticizer Succinic acid can be used as an efficient co- plasticizer in starch based polymer synthesis Blending process is one of the important methods of modifying the polymer characteristics PBS is a biodegradable polyester- synthesized from SA platform

5. Problem statements Most of the research work on Green Technology are patented and details of the invention is not disclosed Currently succinic acid and synthetic polymers in food packaging - produced from petroleum based chemicals - two limitations: non renewability and non biodegradability Typha grass -abundantly available all over the world- not investigated as renewable raw material for high value platform chemical

6. Problem statements PBS synthesized from SA- lacks flexibility. Blending with starch help improve the characteristics of polymer. Glycerol plasticized starch polymer (GTPS) recrystallize on storage- requires a co- plasticizer. Amine co-plasticizer-toxic . Biosuccinic acid - food grade- can serve as an alternative co- plasticizer.

7. Main objective To synthesize bio-succinic acid from Typha grass hydrolysate using microbial fermentation and its application for the synthesis of PBS- starch and SGTPS copolymers

8. Specific objectives To determine the yield of bio- succinic acid produced from Typha grass hydrolysate using A. succinogenes To optimize the ratio succinic acid as co- plasticizer with glycerol to synthesize SGTPS To optimize the ratio of cassava starch and PBS to synthesize copolymerized PBS.

9. Literature review Raw Materials Micro-organisms Result/Findings References wood hydrolysate M. succiniciproducens yield of 56% Batch Fermentation Kim et al. 2004 straw hydrolysate A. Succinogenes 80.7% yield after 48 hrs of fermentation Zheng et al. 2009 cane molasses A. succinogenes yield of 79.5% after 48 hrs of batch fermentation Liu et al. 2008 Production of Succinic Acid by Bacterial fermentation

10. Literature review Raw Materials Polymerization method/ polymer Result/Findings References CA, glycerol, starch CGTPS by melt blending Esterification and better cross-linking Decrease in MW/light Decreased T g Change in crystal structure (reduced retrogradation) Shi et al. 2007 Starch, glycerol, PCL TPS/PCL Blends Decreased Tensile Strength Decreased Elongation at Break Averous et al. 2000 Starch, glycerol, PCL TPS/PCL Blends Decreased Tc Decreased Mp increased % crystallinity Increased Relative crystallinity Huang et al. 1993 Polymer and copolymer synthesis and characterization

11. Materials and Methods Typha grass hydrolysate, Microbial strain pure culture ( A. succinogenes ), sodium/calcium hydroxide, biobased SA, PBS, Cassava starch, Glycerol Materials and chemicals Micro-organisms Actinobacillus succinogenes Anaerobiospirillum succiniciproducens or Mannheimia succiniciproducens

12. Equipments B ioreactor, carbon dioxide cylinder, HPLC with sugar column H igh speed mixer, twin screw co-rotating extruder, Melt Blender, FTIR, NMR, GPC, DSC, SEM

13. METHODOLOGY Biomass (Typha grass) Pre-treatment (Alkali) Hydrolysis (Enzyme/Acid) Fermentable Sugars (Glucose, Xylose ) Hydrolysate Drying, Powdering The method developed by Mr. Idi Audu Guga, an AIT doctoral student, will be followed till hydrolysis Preparation of Hydrolysate Experiment for Objective 1

14. METHODOLOGY contd …… Fermentation ( by A. succinogenes) Product Recovery Precipitation with NaOH 37 C, pH 6.5, Buffer MgCO 3 , 12h, CO 2 Succinic Acid/Sod. Succinate Preparation for Fermentation Fermentation and Product Recovery Experiment for Objective 1

15. METHODOLOGY contd …… Melt Blending SGTPS Characterization Mechanical Biodegradability Physical Co-polymerization and Characterization Extrusion Co-polymerization PBS-starch copolymer PBS Cassava Starch Glycerol + Water Starch + SA High speed blending Experiment for Objective 2 Experiment for Objective 3

16. COMPOSITION of SGTPS Co-polymer Name Abbreviation Sample Weight Proportions Hydrous starch (20% mc wb ) Glycerol SA GTPS SA 100 30 SGTPS 1 SA 1 100 30 1 SGTPS 2 SA 2 100 30 2 SGTPS 5 SA 5 100 30 5 SGTPS 10 SA 10 100 30 10

17. COMPOSITION of PBS-starch Co-polymer Name Sample Weight Proportions Hydrous starch (20% mc wb ) PBS PBS 100 PBSS 1 80 20 PBSS 2 70 30 PBSS 3 60 40 PBSS 4 50 50

18. CHARACTERISTICS TO BE MEASURED Tensile Strength % Elongation at Break IR spectra by FTIR DSC Thermograms Thermogravimetric Analysis Inherent Viscosity (using available viscometer) Degree of Substitution and Esterification (According to Santayanon and Wootthirahokkam , 2003) Biodegradability (using lipase enzyme)

19. Work Plan S.N. Activities Aug Sep Oct Nov Dec Jan Feb Mar Apr 1 Literature review X 2 Procurements of pure culture, chemicals and equipments X 3 Fermentation, Pdt Recovery X X X 4 Blending, Co-polymerization, characterization X X X 5 Result interpretation and data analysis X X 6 Final reporting X X

20. Budget Estimation S.N. Operational Activities Amount (Baht) 1 Chemicals, Pure culture and Enzymes 5000 2 Equipments: (HPLC Column, Melt Blender) 30000 3 Travel 2000 4 Miscellaneous 5000 Total 42000

21. References Bechthold I, Bretz K, Kabasci S, Kopitzky R, Springer A (2008). Succinic acid: a new platform chemical for biobased polymers from renewable resources. Chem Eng Technol 31:647-654. Takiyama , E.; Fujimaki , T. (1994). Bionolle biodegradable plastic through chemical synthesis. In Biodegradable Plastics and Polymers; Doi , Y., Fukuda, K., Eds.; Elsevier Science: Amsterdam, The Netherlands, pp. 150-174. Mochizuki, M.; Mukai , K.; Yamada, K.; Ichise , N.; Murase , S.; Iwaya , Y. (1997). Macromolecules, 30, 7403. Azim, H.; Dekhterman, A; Jiang, Z. and Gross, R.A. (2006). Biomacromolecules, 7, 3093-3097 Shi, R.; Zhang, Z.; Liu, Q.; Han, Y.; Zhang, L.; Chen, D.; Tian, W. (2007). Characterization of citric acid/glycerol co-plasticized thermoplastic starch prepared by melt blending. Carbohydrate Polymers 69, 748–755

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