Microbial Synthesis of SuccinicAcid 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
Main Idea Renewable/Green Chemicals Cheap and Under-utilized resources Biodegradable polymer with better properties
Introduction Succinic Acid is a C4 dicarboxylic acid with molecular formula C4H6O4, molecular weight 118.09 and melting point 185 1900C. 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.
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
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
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.
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
Specific objectives1. To determine the yield of bio- succinic acid produced from Typha grass hydrolysate using A. succinogenes2. To optimize the ratio succinic acid as co- plasticizer with glycerol to synthesize SGTPS3. To optimize the ratio of cassava starch and PBS to synthesize copolymerized PBS.
Literature reviewProduction of Succinic Acid by Bacterial fermentationRaw Materials Micro-organisms Result/Findings Referenceswood hydrolysate M. yield of 56% Kim et al. 2004 succiniciproducens Batch Fermentationstraw hydrolysate A. Succinogenes 80.7% Zheng et al. 2009 yield after 48 hrs of fermentationcane molasses A. succinogenes yield of 79.5% after Liu et al. 2008 48 hrs of batch fermentation
Literature review Polymer and copolymer synthesis and characterizationRaw Materials Polymerization Result/Findings References method/ polymerCA, glycerol, CGTPS by melt • Esterification and better cross- Shi et al. 2007starch blending linking • Decrease in MW/light • Decreased Tg • Change in crystal structure (reduced retrogradation)Starch, glycerol, TPS/PCL Blends • Decreased Tensile Strength Averous et al. 2000PCL • Decreased Elongation at BreakStarch, glycerol, TPS/PCL Blends • Decreased Tc Huang et al. 1993PCL • Decreased Mp • increased % crystallinity • Increased Relative crystallinity
Materials and MethodsMaterials and chemicalsTypha grass hydrolysate, Microbial strain pureculture (A. succinogenes), sodium/calciumhydroxide, biobased SA, PBS, Cassava starch,GlycerolMicro-organisms• Actinobacillus succinogenes• Anaerobiospirillum succiniciproducens or• Mannheimia succiniciproducens
Equipments• Bioreactor, carbon dioxide cylinder,• HPLC with sugar column• High speed mixer,• twin screw co-rotating extruder,• Melt Blender,• FTIR, NMR, GPC, DSC, SEM
Experiment for Objective 1METHODOLOGY Biomass (Typha grass)Drying, Powdering Pre-treatment (Alkali) Preparation of Hydrolysate Hydrolysis (Enzyme/Acid) The method developed by Hydr Mr. Idi Audu Guga, an AIT Fermentable Sugars olysa doctoral student, will be (Glucose, Xylose) te followed till hydrolysis
METHODOLOGY contd……Fermentation and Product Experiment for Objective 1 Recovery Preparation for Fermentation 370C, pH 6.5, Fermentation Buffer MgCO3, ( by A. succinogenes) 12h, CO2 Product Recovery Precipitation with NaOH Succinic Acid/Sod. Succinate
METHODOLOGY contd……Co-polymerization and Characterization Glycerol + Cassava Starch + SA PBS Water Starch High speed blending Experiment for Objective 3 Extrusion Co-Experiment for Objective 2 polymerization Melt Blending SGTPS PBS-starch copolymer Characterization Physical Mechanical Biodegradability
COMPOSITION of SGTPSCo-polymer Abbrevi Sample Weight Proportions Name ation Hydrous Glycerol SA starch (20% mc wb)GTPS SA0 100 30 0SGTPS1 SA1 100 30 1SGTPS2 SA2 100 30 2SGTPS5 SA5 100 30 5SGTPS10 SA10 100 30 10
COMPOSITION of PBS-starchCo-polymer Name Sample Weight Proportions Hydrous starch (20% PBS mc wb)PBS 100 0PBSS1 80 20PBSS2 70 30PBSS3 60 40PBSS4 50 50
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)
Work PlanS. Activities Aug Sep Oct Nov Dec Jan Feb Mar AprN.1 Literature review X2 Procurements of X pure culture, chemicals and equipments3 Fermentation, X X X Pdt Recovery4 Blending, Co- X X X polymerization, characterization5 Result X X interpretation and data analysis6 Final reporting X X
Budget EstimationS.N. Operational Activities Amount (Baht)1 Chemicals, Pure culture and Enzymes 50002 Equipments: 30000 (HPLC Column, Melt Blender)3 Travel 20004 Miscellaneous 5000 Total 42000
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