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S-1007 Multistate Research Committee
The Science and Engineering for a Biobased Industry and Economy
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Raw Materials, Byproducts,
Agricultural “wastes”
Bioprocessing,
Bioconversion,
Bioreactor design and
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control, “Biorefineries”
Bulk
Chemicals,
Biofuels
Specialty
Biomaterials
Biochemicals
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Biochemicals Subgroup
Donal Day,
Shulin Chen,
Mark Eiteman,
Arun Bhunia,
Mike Tumbleson,
Caye Drapcho,
Danielle Carrier,
Cady Engler,
Jin-Woo Kim,
Ruihong Zhang,
Lorraine Day,
Mike Zhang,
Lonnie Ingram,
Terry Walker,
John Benemann,
Any others?
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Local
power grid
•Electricity
•Electricity
•Steam
•Steam
•Electricity
•Steam
System Boundary
Inputs
•Fertilizers
•Fuel
•Agrochemicals
Cropping
systems
Animal
Animal waste
treatment
•Nutrient
operation
Energy Inputs
Surplus
•Electricity
Inputs
•Steam
•Chemicals, Enzyme
Biorefinery
Biorefinery
Pre -Processing
Intermediates
•Sugars
•Lipids
•Lignin
•Ash
•Protein
Final product
•Food
•Food
•Other
•Otherproducts
products
Final Processing
Final product
•Ethanol
•Ethanol
•Biopolymers
•Biopolymers
•Chemicals
•Chemicals
•Hydrogen
•Hydrogen
Cogeneration
•Ash
Biopharmaceutical Mechanisms
• Macro
– Whole animal system
– Whole plant system (glycosylation limitation)
– Whole insect system ( “ )
• Micro
–
–
–
–
Mammalian Cell Culture
Plant cell culture
Insect cell culture (baculovirus mechanism)
Bacterial and fungal culture (E. coli, S.cerevisiae, Pichiae)
• Molecular
– Genomics, Proteomics, Metabolomics, Systems Biology
– Chaperone mechanisms
Byprodu ct
Primar
y
Primar y enzyme
Product
Byproduct
Bioconversions
to
Products
of Increasing Value
Substrate
/Microbi al strain
Animal waste
Wood fibers
Potato/ S weet
potato
Dairy waste
Oily waste
Rice brokens
Rice straw
Switchgrass
Complex
Xylan
Glucose
Sugarcane
bagasse
Corn stover
Xylose
Pyruvate
Acrylonitril
e
D-xylulose
Corn starch
Glucose
Molasses
(sugarcane)
Sucrose
Lactose
Lipids
Glucose
Xylose
Xylose
Glucose
Glucose
Glucose
Glucose
Glucose
Glucose
Specific
Media
Glucose
Glucose
Glucose
Glucose
Serum
Serum
Serum
(M. flagellate)
Clostridium sp. SAIV1
Alcohol dehydrogenase
(K. marxianus)
-galactosidase
Lipase (esterases)
(C. acetobutylicum)
Citrate synthase
Lactose dehydrogenase
(L. delbrueckii)
Pyruvate decarboxylase
(Acetobacter sp.)
Nitrile hydratase
Methane
Ethanol
Ethanol
(thermophilic pathway)
Glucose and Galactose
Fatty acids
Acetone/butanol (30/60)
Citric acid
Lactic acid
(30 million kg/yr)
Acetic acid
Xylose reductase (C.
tropicalis)2
Glucose isomerase
Xylitol
Fumarase
(B. ammoniagenes)
AcetylCoA carboxylase
(C. glutamicum)-biotin
Aspartic amino
transferase (E. coli)
Aspartase (E. coli)
L-aminocaprolactam
hydrolase (C. laurentii)
(P. fluorescens)
Penicillin amidase
(P. chrysogenum)
(B. lichenformis)
(B. amyloliquefaciens)
(Rhizopus, Aspergillus)
Anthrobacter simplex
Hybridomas
Human fibroblasts
Monkey kidney cells
1 Murty and Chandra (1997)
2 Hortis u et al. (1992)
Acrylamide (15,000 tons/yr)
High fructose corn syrup (8
million tons/yr)
L-malic acid
L-glutamic acid (340,000
tons/yr) MS G
L-phenylalanine
(Aspartame synthesis)
L-aspartic acid
L-lysine (70,000 tons/yr)
($2/lb)
L-histidine
6-aminopenicilloic acid (7500
tons/yr)
Proteases ($236 mil/yr)
Amylases ($70 mil/yr)
Other enzymes ($92 mil/yr)
Prednisolone
Monoclonal antibodies
Interferon
Polio vaccine
Increasing
Product
Value
Examples of Research
University Collaborations
Research at Clemson University
• Fungal production of essential fatty acids
and SFE fractionation
– Walker, Hui, Kuan, Heaton, Cantrell
• Algal production of polar lipids and
proposed hydrogen co-production
– Drapcho, Brune, Walker
• “Microalgae Platform”
– Brune, Benemann
Bioconversion and SFE Process
SFE
i. Raw Material(s)
ii. Organism Selection
iii. Metabolic Engineering
Microfiltration
Monitor
-Control
Station
Bioreactor
Partially-Refined
Product
Purified Product
LC and
Analysis
(HPLC or GC)
Figure 1. Typical bioprocess for bioconversion of raw materials to nutraceutical and specialty
biochemicals. Primary separation procedures include ultrafiltration (UF), supercritical fluid
extraction (SFE) and liquid chromatography (LC). Analytical techniques include gas
chromatography (GC) and high pressure liquid chromatography (HPLC).
Samples of oil extracted by different extraction methods
compared to commercial sample of refined rice bran oil
Soxhlet extracted
sample
(biotreated bran)
Soxhlet extracted
sample
(raw bran)
SFE extracted
sample
(biotreated bran)
Commercial
refined rice ban
oil
Figure 1. Conceptual Process Flow Diagram for Hydrogen Production
from Biomass
Enzyme
Hydrolysis &
H2 Production
AFEX
Pretreatment
Biomass
H2 + CO2
Novel
Membrane
Separation
Novel CO2-fixing
Microbial Strains
Co-products
(Organic acids)
Methane-forming
Microbial Cultures
CO2
Absorption
Steam
Reforming
Methane
Production CH4 + CO2
CH4
CO2
Absorption
Pure
H2
Energy
Research at
Washington State University
• Manure conversion to sugars:
– Analysis of manure components: cellulose,
hemicellulose, lignin, sugar, protein, amino acids, and
metal element,
– Extensive studies were conducted on pretreatment
•
•
•
•
single stage acid hydrolysis, two-stage acid hydrolysis,
dilute acid hydrolysis, concentrated acid hydrolysis,
enzyme hydrolysis
combined acid and enzyme hydrolysis.
• Cull Potato conversion to lactic acid
– Fungal and bacterial fermentation
• Whey/whey permeate conversion to nisin
Research at Virginia Tech
• Protein separation from feedstocks
– Aqueous two-phase extractions
– Zhang
• Xylitol Bioproduction
Research at LSU and
University of Arkansas
• Low-cost pretreatment of bagasse
– Delignification using singlet oxygen
– Donal Day (LSU Audubon Sugar Institute)
• Characterization and extraction of
antioxidants in blackstrap molasses
– Julie Carrier (UARK), D. Day
?s