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Fermentation of pretreated source
separated organic (SSO) waste for
ethanol production by different
bacteria by
Bekmuradov Valeriy,
Luk Grace and
Luong Robin
Ryerson University
Toronto, Canada
Montreal, Canada 2013
1
Presentation Outline
• Introduction
• Objectives
• Methodology
• Experimental plan
• Results
• References
2
Introduction
• History of the study – waste problem,
energy crisis, pollution?
• Ethanol? Reasons for use?
• Feedstock availability?
• Processing steps:
– Pre-treatment (Thermal Screw Press)
– COSLIF pre-treatment
– Enzymatic hydrolysis
– Fermentation
3
Ethanol use today
• Used extensively in
some areas of world
– Brazil
• leading user
– United States
• ethanol use is still small %
• U.S. is leading producer
– 6,503.6 million gallons
• Most gasoline engines
can run on E10
U.S.A.
Brazil
E.U.
China
Canada
Other
1.6%
Annual Fuel Ethanol Production (2011)
38.3 %
49.6 %
4
More reasons to use ethanol
• Energy security
• Environmental concerns
• Foreign exchange savings
• Socioeconomic issues related to rural
sector
5
However…today
• Agriculture can have a profoundly positive or negative impact on soil and water quality, water and land use, habitat
• Ethanol from agricultural crops (food based biomass) are still expensive ($2.7 per gallon)
• Needs to occupy new land to produce enough amount of ethanol to meet demand (corn production for ethanol compete with food for land needed)
• Feedstock has uneven fluctuation during the year
• Emission of GHG from food based ethanol is high
6
What is next?
One option is…
• Cellulosic ethanol:
– derived from lignocellulosic biomass • structural component
• i.e. wood, corn stover (leaves and stalks), grasses
• Advantages:
– abundant sources
– could reduce greenhouse gas emissions by 85%
• Disadvantages:
– require more processing to get usable glucose
7
Objectives
• Investigate pretreated SSO waste for sugar and ethanol yields by separate hydrolysis and fermentation (SHF) approach
• Study on performance of commercial available enzyme complex - Accellerase 1500
• Evaluation of selected hydrolysate by fermentation with different bacteria - Z. mobilis 8b and S. cerevisiae DA2416
• Propose a low-cost method utilizing waste biomass for ethanol production and other valuable products
8
Methodology
• Feedstock Material: Source Separated Organic (SSO) waste + Construction & Demolition (CD) wood waste
• Aufbereitungs Technology and System (ATS) thermal screw machine: High pressure and high temperature along with screwing makes the SSO to be a fibrous, homogenous, and less odorous material • Sampling Procedure: Method of Jansen et al. 2004 for obtaining a representative sample • Measurement Procedures: NREL, ASTM, and TAPPI
9
Lignocellulosic biomass
• Introducing Lignocellulosic Biomass resource instead of food-based biomass such as corn and rice
• Lignocellulose is one of the most abundant resources on the Earth on negative cost
• Cellulose: Linear, Insoluble biopolymer composed of repeated unions D-glucose units bonded by ß 1-4 linkages
• Can be hydrolysed to glucose by cellulase enzymes from some microorganisms
10
Lignocellulosic biomass
• Hemicellulose: Random, amorphous, branched chains structure composed of pentose, xylose, other sugars, easily hydrolyzed by dilute acid, base, and hemicellulase enzymes
• Lignin: Complex, three-dimensional polymer of polyphenolic compounds in branched chains, non-crystalline and its structure is similar to a gel or foam
11
Major processing steps in biomass
conversion
12
Pretreatment
13
Pretreatment
14
Deconstruction of plants
15
Termochemical route: courtesy of
DOE/NREL
Biomass composition
16
Biomass composition
17
Mass balance of SSO
18
Limitation of Lignocellulosic
biomass
• Presence of Lignin
• Cellulose Crystallinity
• Accessible Surface Area
• Acetyl Content
• Presence of Hemicellulose
• Almost all lignocellulosic biomass materials need pre-treatment. Without pre-treatment the hydrolysis yield can barely exceed 20% of theoretical yield whereas yields after pre-treatment can reach up to 90% (Lynd, 1996).
19
Experimental plan
20
Parameters of interest
• Pretreatment: temperature, pH, pressure
• Enzymatic hydrolysis: enzyme loading, reaction conditions, substrate concentration, substrate particle size, adsorption capacity and cellulose hydrolysis rate constants, sugar yields
• Fermentation parameters:
ethanol yields & concentration.
21
COSLIF pre-treatment steps
22
COSLIF pre-treatment
23
COSLIF pre-treatment
24
Structural difference on avicel as a pure cellulose (A and B),
avicel treated by 77% phosphoric acid (C and D), treated by
83% phosphoric acid (E and F). Similarly, corn stover substrate
(G) before pretreatment and (H) after COSLIF pretreatment with
85% phosphoric acid.
Enzymatic Hydrolysis Results
25
Glucose yield after Enzymatic Hydrolysis (37°C for 48 hours, pH=4.8, FPU=30)
COSLIF Std – washing with acetone
COSLIF Mod – washing with ethanol
0
10
20
30
40
50
60
70
80
90
100
COSLIF Std COSLIF Mod
COSLIF Mod
COSLIF Std
Glucose Yield
Yie
ld %
Enzymatic Hydrolysis Results
Glucan digestibility profiles from COSLIF Std (FPU=60, acetone)
and COSLIF Mod (FPU=30, ethanol) pretreated samples
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0
10
20
30
40
50
60
70
80
90
100
0 12 24 36 48 60 72
COSLIF Std
COSLIF Mod
Glu
ca
n d
ige
stib
ility
%
Time (hr)
Fermentation results
using Z.mobilis 8b strain;
using S.cerevisiae strain
27
0
20
40
60
80
100
120
6hr 12hr 24hr 48hr
COSLIF- Z.mob
COSLIF - S.cer
Ethanol Yield
Th
eo
retical e
tha
no
l yie
ld, %
Fermentation results
140 g/L is equivalent to 0.48 g ethanol/ g biomass
28
0
20
40
60
80
100
120
140
160
6hr 12hr 24hr 48hr
COSLIF- Z.mob
COSLIF-S.cer
Ethanol Conc.
Eth
an
ol,(g
/L)%
Fermentation results
Ethanol yield comparison
29
Theoretical Ethanol Yield from SSO
Total ethanol yield from 1 ton of dry SSO: 171.3 + 93.95 = 265 L
City of Toronto collects approximately 100,000 tons of SSO per year.
Assuming 45% of the dry weight of SSO, 45,000 tons of dried SSO per
year is available in the city of Toronto, from which ~12ML litres of
ethanol can be produced.
30
Advantages of biofuel to replace
gasoline
• Cellulosic bio-fuels can displace 8 million barrels of oil per day- equal to all of the oil used by light-duty vehicles today.
• Bio-fuels can be second only to vehicle fuel economy improvements in the amount of oil they save.
• Bio-fuels, vehicle efficiency and smart growth could eliminate virtually all our demand for gasoline.
• Bio-fuels could reduce global warming pollution by 1.7 billion tons per year-23% of total U.S emissions in 2012.
31
Improving efficiency in biofuel
production
• By 2050, demand rises from the current 160 billion gallons to 289 billion gallons.
• To meet all of this with current crops and current cellulosic conversion technologies, it would required over 1.8 billion acres of land.
• Readily, achievable advances in vehicle fuel economy, overall transport efficiency, crop yields and conversion efficiency could reduce the land requirement to just 116 million acres.
32
Future trends
• Ethanol derived from the cellulosic part of plants rather than just the starch, are the most promising fuels for the transportation sector.
• Replacing oil with bio-fuels would allow to reinvest billions of dollars in factories & farms.
• To maximize the benefits from bio-fuels, need to push technology & market to develop quickly.
33
References • Lynd, R. L., Elander, R. T., & Wyman, C. E. (1996). Likely features and cost of
mature biomass technology. Appl. Biochem. Biotechnol. 57/58: 741-761.
• McMillan, J. D. (1994). Pretreatment of lignocellulosic biomass. In: Himmel, M. E., Baker, J. O., Overend, R. P., (Eds). Enzymatic conversion of biomass for fuels production. American Chemical Society, Washington, DC, 292-324.
• Mirzajani, M. (2009). “The amenability of pre-treated source separated organic (SSO) waste for ethanol production”. Master’s thesis, Ryerson University, Civil Engineering dept., Toronto, Canada.
• Ehsanipour, M. (2010).“Acid pretreatment and fractionation of source separated organic waste for lignocellulosic sacharification”. Master’s thesis, Ryerson University, Civil Engineering Dept., Toronto, Canada.
• South, S.R., Hogsett, D., & Lynd, L. (1995). Modeling simultaneous saccharification and fermentation of lignocellulose to ethanol in batch and continuous reactors. Enzyme Microb Technol. 17:797-803
• Vartek Ltd. (2005). Vartek ATS Technology Compost Pilot test. Toronto, Ontario, Canada, Vartek Company.
• Wyman, C. E. (1999). Biomass ethanol: Technical progress, opportunities, and commercial challenges. Annu Rev Energy Environ 24: 189-226.
• Zhang, J., Shao, X., Townsend, O.V., & Lynd, L.R. (2009). Simultaneous saccharification of paper sludge to ethanol by Saccharomyces cerevisiae RWB222. Biotechnology and Bioengineering, 104(5), 920-931.
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Thank you
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