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Biofuels Symposim
- 1. Enzymatic Hydrolysis of Pulp in N-Methylmorpholine–N-Oxide/WaterBrett RobbinsFAMU-FSU College of EngineeringDepartment of Chemical and Biomedical Engineering
- 2. Traditional Biomass to Ethanol Schematic - DOE report
- 3. - DOE report
- 4. Solvent should have a good capacity for dissolution of cellulose Important to pick a solvent which does not inactivate enzymes. - DOE report
- 6. Excellent solvent for dissolution of cellulose
- 7. Environmentally friendly UN award for green process
- 9. No need to form cellulose derivatives- J. Collier and co-workers NMMO interferes with the hydrogen bonds of cellulose and breaks up the crystalline structure
- 11. But also interferes with H-bonds in cellulases deactivation1-butyl-3-methylimidazolium chloride [Bmim]Cl - Rogers and co-workers 1-ethyl-3-methylimidazolium acetate [Emim]Ac 1-ethyl-3-methylimidazolium diethyl phosphate [Emim]DeP - Kamiya and co-workers Is in-situ hydrolysis possible in these solvents?
- 13. To develop a continuous process using twin screw reactor for processing of high lignocellulosic biomass solutions
- 14. Enzymatic solution phase hydrolysis in twin screw reactor
- 15. Conversion to simple sugars
- 16. Agricultural residues and processing byproducts
- 18. Are the rate of reactions higher in the dissolved phase than in the regenerated phase?
- 19. How do reaction rates vary as a function of solution conditions?
- 20. Temperature, Enzyme Loading, pH
- 21. Which solvent produces a higher yield of sugars?
- 22. Is the extruder capable of supporting a reacting recycle system?Answering these questions will help establish experimental conditions to develop a continuous process.
- 23. Method of Assay: DNS 3,5-Dinitrosalicylic acid (DNS or DNSA) is an aromatic compound that reacts with reducing sugars and other reducing molecules to form 3-amino-5-nitrosalicylic acid, which absorbs light strongly at 540 nm. A reducing sugar is any sugar that, in basic solution, forms some aldehydeor ketone. Reducing sugars include glucose, fructose, glyceraldehyde, lactose, arabinose and maltose. - Miller, 1959 Increasing sugar concentration
- 24. Characterizing the Cellulase Enzymes are characterized in terms of Filter Paper Units – FPU FPU is defined as the enzyme that releases 1 µmol of glucose equivalents per minute from a Whatman No. 1 filter paper. Accellerase 1000: 56 FPU/ml
- 25. Experimental Conditions Cellulose Solutions Reaction Conditions
- 28. Higher Cellulose Loading pH = 5.7; T = 50o C; Enzyme Loading = 122 FPU/g Cellulose 2.31 wt% Similar trends at higher cellulose loadings
- 31. Effect of pH on of Reaction Rate T = 50o C; Enzyme Loading = 122 FPU/g Cellulose 2.31 wt% At 50°C, Rates and final cellulose conversion are higher at a pH of 5.75 at all enzyme loadings studied
- 32. Comparison between NMMO and Ionic Liquid 7.22 mg/mL cellulose loading, T = 50o C; Enzyme Loading = 122 FPU/g Cellulose NMMO Ionic Liquids [Emim]Ac [Emim]DeP Solvent effects on in-situ enzymatic hydrolysis NMMO > [Emim] DEP > [Emim] Ac >> [Bmim] Cl Does anion in the ionic liquid inactivate cellulases? How to predict enzyme activity in different solvents ? – Hildebrand solubility parameters, dielectric constants, octanol-water partition coefficient – ongoing work.
- 34. Enzymatic Hydrolysis at Higher Solids Loading T = 50o C; Enzyme Loading = 122 FPU/g Higher reducing sugar yield However Percent conversion are similar
- 38. Sun Grant “US DOT Sun Grant, Enzymatic and Multiphase Solution Processing of Lignocellulosic Biomass.
- 39. Bush Brothers & Company, Industrial Support for Biomass Research.
- 40. Florida State University Undergraduate Honors Thesis Award
Editor's Notes
- Traditional Biomass Flow ChartPretreatment Step requires two subsequent steps to prepare the solution for reaction (explain Acid or IL)If we can create a single phase in which pretreatment, hydrolysis and fermentation can occur, flow chart greatly simplified
- This is the goal process schematic of the Department of Energy3-Step plan - Increase Agriculture efficiency, Improve Enzyme Complex, Combine Hydrolysis and Fermentation StepsWill reduce production costs which reduces costs to the consumerTo do this we will need to take a closer look into the individual steps
- In the Pretreatment Step the lignocellulosic structure is broken downThe cellulose polymer chains are held together by hemicellulose and ligninBy interfering with the hydrogen bonds that hold these structures together, the individual components can be exposed for hydrolysisHowever, a solvent that is capable of dissolving lignocellulose must also not interfere with the enzyme activityThere is a commercially viable solvent available, but is it more effective than the more novel Ionic Liquids?
- N-methylmorphiline-N-oxide (NMMO) is a commercially available solvent used to dissolve cellulose in the Lyocell process [describe Figure]NMMO can be found in 4 different complexes, however monohydrate has the necessary properties to dissolve celluloseIt is at the tertiary nitrogen where a strong dipole results in an electronegative oxygen [point]This oxygen interferes with the hydrogen bonds of cellulose to facilitate dissolutionNot only can it dissolve cellulose, but NMMO has a low vapor pressure which permits high recovery
- Do not like slide
- Cellulase complexes are made up of 3 classes of enzymes- Endoglucanase, Exoglucanase and β-glucosidaseEach type plays a role in the breakdown of cellulose polymer chainsCommercially developed enzyme complexes are specifically designed to hydrolyze celluloseFor this study Accelerase™ 1000 obtained from Genencor were usedUsed alpha cellulose which is a dissolving pulp with DP 1160, where most lignin and hemicellulose is removed was obtained from Buckeye Technologies, Perry FL
- Go through description of goals and thought process-a few questions arise that need to be answered in order to reach these goals
- Go though questions – enzymes active? In situ or regenerated? Conditions (pH, Temp, Enzyme)? Cellulosic solvent? Recycle system?
- To determine reducing sugar yields, the DNS assay was utilized3,5 dinitrosalicyclic acid reacts with the aldehyde or ketone of a reducing sugar producing an amine group The molecule absorbs light in the visible spectrum and can be anaylzed using a UV-vis spectrometer (point to image)
- Because the exact contents of commercial enzymes are trade secretsFilter Paper Units can be used to determine enzyme activity(just brief over)
- NMMO/Cellulose solutions were made at 1, 2.31, 5 and 10 wt% cellulose in NMMOAfter preparation of he reaction solutions the initial cellulose concentrations were 3.1,
- Describe reaction conditions (1 wt% cellulose = 3.13 mg/mL), then Graph (Clearly)Red? What is BLUE??
- All are 2.31 wt% cellulose?? Different information???
- Even though pH is high we still get 50% conversion when cellulose is dissolved in solution. Harmful effects of solvent and pH are overcome when cellulose is dissolved in solution2.31% cellulose
- (more comments)Modification of a Rheomax OS extruder to provide recycle systemWas able to produce 5 and 10 wt% cellulose solutions
- Most of the work presented today is done by Rilwan and Brian