Platform Molecules and Green Chemistry
Platform Chemicals From
Renewable Raw Materials
Professor Colin Webb
Satake Centre for Grain Process Engineering
School of Chemical Engineering and Analytical Science
The University of Manchester
Renewable Resources & Biorefineries Conference, September 6–8, 2006, York
Raw Materials, Agriculture and Chemistry
“Petrochemical processes have provided low
cost production routes for fuels, plastics and
chemicals for well over 50 years but the
escalating impact on the environment and the
inevitable future depletion of fossil
feedstocks make it essential that benign,
sustainable, alternatives be developed
commercially in the very near future.”
RSC, 2003
RRB2, York, September 2006
Renewable raw materials
zPhotosynthetic biomass
¾Starchy crops
¾Sugar crops
¾Oil seeds
¾Lignocellulosics
zWater
zAtmospheric gases (e.g. CO2, methane)
RRB2, York, September 2006
Platform chemicals from petroleum
Heavy fuel oil
Acetic acid
6 ?109 kg
$0.99/kg
Methanol
29.1?109 kg
$0.21/kg
Terephthalicacid
3.2?109 kg
$0.63/kg
Formaldehyde
4.9?109 kg
$0.46/kg
Natural gas liquids
(methane, ethane,
propane)
o-/m-/p-xylenes
10.1?109 kg
$0.31/kg
Crude oil
Propylene
43.1x 109 kg
$0.44/kg
BTX n-Butanols
1.78?109 kg
Cumene
8?109 kg
$0.56/kg
Acetone
9
Cyclohexanone, 3.4?10 kg
$0.46/kg
Nylon-6,6
Styrene
19.2?109 kg
$0.55/kg
Acrylic fibres,
nitrilerubber,
ABS, SAN
Polystyrene,
SBR, ABS,
SAN, resins
MTBE
22.2 x109 kg
$0.29/kg
Polyethylene
<$1/kg
Vinyl acetate
3.8?109 kg
$1.1/kg
Propylene
glycol
0.97?109 kg
$1.48/kg
Butadiene
8.5 x109 kg
$0.57/kg
Mixture
Butane/enes
Polypropylene
<$1/kg
Ethylbenzene
19.7?109 kg
$0.56/kg
Phenol
3.4?109 kg
$0.82/kg
Oil coke
Ethylene
79 x109 kg
$0.6/kg
Isopropanol Acrylic acid Acrylonitrile Propylene
oxide
2 ?109 kg
2.3?109 kg
4.3?109 kg
$0.84/kg
$1.92/kg
$1.98/kg
4 ?109 kg
$1.41/kg
Diisocyanates
2.23?109 kg
$2.2/kg
Polybutadiene
1.82 x109 kg
Light fuel/diesel
oil
Naphtha
BTX
Toluene
Benzene
7.85?109 kg 20.79?109 kg
$0.33/kg
$0.33/kg
Cyclohexane
5.1?109 kg
$0.44/kg
Jet fuel
Ethylene oxide
11.2?109 kg
$1.17/kg
Ethanolamines
0.84?109 kg
$1.28/kg
RRB2, York, September 2006
Vinyl
chloride
24?109 kg
$0.48/kg
Ethylene glycol
9.7?109 kg
$0.53/kg
Ethanol
2.6?109 kg
$0.42/kg
Polyvinyl
chloride
New products or new processes?
Sustainable
Raw Material
New Process
Conventional
Product
Replacement
Sustainable
Raw Material
Conventional
Process
Alternative
Products
Sustainable
Raw Material
New Process
Alternative
Products
RRB2, York, September 2006
Bioprocessing
z Bioprocessing will lead to more natural and cost-effective
means of converting natural feedstocks into platform
chemicals
z Platform chemicals that can be produced in large quantities by
microorganisms include acetic acid, ethanol, glycerol, succinic
acid, pyruvic acid, 2,3-butanediol and 1,3-propanediol
z They are different from current petrochemical building blocks
(e.g. ethylene, propylene and butadiene) and therefore require
significant technological research
z Benefit will come from collaboration of chemists,
biotechnologists and (bio)chemical-engineers
RRB2, York, September 2006
Cereals as a Renewable Resource
z Sustainable renewable chemicals will have to come
mostly through photosynthesis
z The most intensive crops are those based on starch
(10 times more chemical energy is stored as cereal
grain starch than as sucrose)
z 50% of the world’s food is cereal based
z Cereal processing for human consumption leaves
approximately 15% of the grain as waste product
z Large amounts of cereal grain are also wasted
through poor post-harvest techniques
z and…yields are still increasing
RRB2, York, September 2006
Cereal Grain or Whole Crop?
z For every tonne of grain most cereals produce
approximately one tonne of straw and other residues
z Ligno-cellulosic material is much more difficult to
process than starchy material
z Straw has a bulk density 1/15th of that of grain
z Biorefinery processing of straw is currently not
feasible
z Like petroleum refining we should start simple
(glucose cf. gasoline; lignocellulose cf. bitumen)
RRB2, York, September 2006
Chemical Products From Glucose
2,3-BUTANEDIOL
LACTIC ACID
Glycerol
BUTANOL
SCP
SORBITOL
POLYBUTADIENE
ACRYLATE RESINS
PVC RESINS
DIBUTYL PHTHALATE
ACETIC ACID
ETHANOL
BUTADIENE
POLYCARBONATE RESINS
BISPHENOL A
METHYL iso-BUTYL KETONE
METHYL METHACRYLATE
ACETONE
GLUCOSE
BUTANEDIOL ACETATE
METHYL ETHYL KETONE
METHYL VINYL KETONE
DIMETHYL TEREPHTHALATE
ACETIC RESINS
ETHYLENE
ETHANOLAMINES
BUTADIENE
DRUGS
POLYURETHANES
SURFACTANTS
HYDROXYMETHYLFURFURAL
POLYETHYLENE
VINYL ACETATE
STYRENE
ACETALDEHYDE
ETHYLENE GLYCOL
TEREPHTHALIC ACID
RRB2, York, September 2006
Top 12 Sugar-based Building Blocks
1,4 succinic, fumaric and malic acids
2,5-furan dicarboxylic acid
3-hydroxybutyrolactone
3-hydroxy propionic acid
Aspartic acid
Glucaric acid
Glutamic acid
Glycerol
Itaconic acid
Levulinic acid
Sorbitol
Xylitol/arabinitol
RRB2, York, September 2006
Succinic Acid
z C4H6O4; MW = 118.09
z Butanedioic acid; 1,2-ethandicarboxylic acid;
dihydrofumaric acid and amber acid
z Important intermediate in the manufacture of:
¾ pharmaceuticals, elastomers, protective coatings, electrical
insulations, foods, detergents, surfactants, corrosion
inhibitors and cosmetics
¾ with a total market size of more than $400,000,000 p a.
z Currently produced from petrochemical feedstocks
(butane - maleic anhydride - succinic acid)
z Bioproduction could decrease production cost from
$7/kg to $0.6/kg
RRB2, York, September 2006
Succinic Acid Derivatives
O
O
OH
HO
1,4-Butanediol (BDO)
Solvent
γ-Butyrolactone (GBL)
Solvent
O
O
H2N
Succindiamide
PBS Polymer
Tetrahydrofuran (THF)
Solvent
O
H
N
2-Pyrrolidone
Solvent
O
HO
OH
NH2
H2N
O
1,4-Diaminobutane
Polyamide 46 (Nylon)
Succinic acid
CH3
O
N H2
O
N
N-Methyl Pyrrolidone (NMP)
Solvent
CN
O
H3C
O
O
CH3
O
Dibasic Ester (DBE)
Solvent, polymer
RRB2, York, September 2006
CN
Succinonitrile
Resin
Bioconversion of glucose to Succinic acid
Actinobacillus succinogenes:
z Gram-negative facultatively anaerobic bacterium
isolated from bovine rumen in 1999
z Moderate halophile & osmophile
z Capnophilic & aerotolerant
z Can use both pentose and hexose sugars
z Natural succinate producer
z Produces succinate mainly via a single pathway
z Consumes CO2 as part of the production process
RRB2, York, September 2006
Metabolic Pathway
Glucose
Glucose-6P
4H
CO2 in
OAA
PEP
out
Acetyl-coA
Pyruvate
CO2 2H
2H
Malate
Acetate
2H
Fumarate
Citrate
Succinate
out
4H
out
2H
Lactate
Ethanol
out
CO2
α-Ketoglutarate
Out
RRB2, York, September 2006
H: reducing power
Metabolic Pathway
Glucose
Glucose-6P
4H
CO2 in
OAA
PEP
out
Acetyl-coA
Pyruvate
CO2 2H
2H
Malate
Acetate
2H
Fumarate
Citrate
Succinate
out
4H
out
2H
Lactate
Ethanol
out
CO2
α-Ketoglutarate
Out
RRB2, York, September 2006
H: reducing power
Current raw material - Medium 1
Glucose
Yeast Extract
B12
Biotin
Folic acid
Riboflavin
Niacin
Pantothenate (B5)
p-aminobenzoate
Thiamine
Pyridoxine (B6)
NaCl
NaH2P4∙H2O
Na2HP4
CaCl2∙2H2O
MgCl2∙6H2O
Vitamins
Minerals
RRB2, York, September 2006
US Patent
5,573,931
Bioconversion of Cereals to Succinic Acid
Upstream
processing
Bioreaction
Wheat
Derivatives
RRB2, York, September 2006
Downstream
processing
Succinic
acid
Bioconversion of Cereals to Succinic Acid
Wheat
contains all nutrients generally required for bioprocessing
Upstream
Downstream
Bioreaction
processing
processing
Succinic
Macronutrients
Micronutrients
Vitamins
-1
-1
-1acid
(µg g , db)
(µg g , db)
(µg g , db)
Starch 620000 - 750000
N
2006
P
3400
S
912 - 1995
Mg
1589
Fe
37
K
4180
Ca
390
Derivatives
Na
Cu
Cd
Cr
Pb
Ni
Mn
Sn
Zn
RRB2, York, September 2006
30
4.5
0.1
0.4
1
0.3
35
7.9
5
B1
B2
B3
B6
E
5.4
1.1
110
3.3
54
Bioconversion of Cereals to Succinic Acid
Wheat
contains all nutrients generally required for bioprocessing
Upstream
Downstream
Bioreaction
processing
processing
Succinic
…butMacronutrients
not in a directly accessible
form:
Micronutrients
Vitamins
-1
-1
-1acid
(µg g , db)
(µg g , db)
(µg g , db)
‰ The carbon source is mostly in the form of
Starch
620000 - 750000
Na
30
B1
5.4
starch
N
2006
Cu
4.5
B2
1.1
¾ most fermentations require simpler sugars
P
3400
Cd
0.1
B3
110
S
912 - 1995
Cr
0.4
B6
3.3
‰ Likewise most of the Nitrogen is in the form of
Mg
1589
Pb
1
E
54
proteins
Fe
37
Ni
0.3
¾ most
fermentations
K
4180
Mnrequire35free amino acids
Ca
390
Sn
7.9
‰ Some pre-processing
required
Znis therefore
5
Derivatives
RRB2, York, September 2006
Upstream Processing
Wheat
Upstream
processing
Bioreaction
Downstream
processing
B2
Succinic
acid
6
10
7
8
9
Hydrolysis
Wheat
Flour
Flour
Hydrolysate
Filtrate
Generic feedstock
Fungal
fermentation
Fungal
Autolysate
Solid
Aspergillus
awamori
Glucose
rich
Autolysis
RRB2, York, September 2006
Nitrogen
rich
Upstream processing
Generic feedstock
provides
C source
N source
Succinic acid
fermentation requires
Hydrolysate
Glucose: 335 g/L
Glucose: 50 g/L
Autolysate
FAN: 0.925 g/L
(Equiv. to 18.5 g/L
Yeast Extract)
Yeast Extract: 5 g/L
RRB2, York, September 2006
Bioreaction
Wheat
Upstream
processing
Bioreaction
Downstream
processing
B2
Succinic
acid
6
10
7
8
9
Flour
Hydrolysate
Cell
Autolysate
slope
Generic
feedstock
A. succinogenes
inoculum
NaOH
Broth
B2
6
10
7
8
CO2
9
RRB2, York, September 2006
Bioreaction
Small anaerobic bioreactors with
1 L bench-top bioreactor
gas distributor
CO2 feed in
pH control
Temperature control
Shaking
Sterilisation
RRB2, York, September 2006
Batch fermentation
7
45
40
6
5
30
25
4
20
3
15
OD660
Concentration (g/L)
35
Glucose
Succinic
OD660
2
10
1
5
-
30
0
5
10
15
20
Ferm entation tim e (hours)
25
25
30
Concentration (g/L)
0
20
Pyruvic acid
Succinic acid
15
Formic acid
10
Acetic acid
5
0
10
20
Fermentation Time (hours)
RRB2, York, September 2006
30
Summary of Batch Bioreactions
No.
Glucose
Vitamins
pH
control
OD660
SA
Product
-ivity
(g/L)
(g/L/h)
Yield
Initial
Consumed
(g/L)
(g/L)
Y/N
Y/N
1
44*
44
N
N
5.5
0.28
0.01
0.01
2
58
30
N
N
1.39
0
0.00
0.00
3
50
15
N
N
2.65
6.15
0.12
0.41
4
30
18
N
N
1.79
1.09
0.03
0.06
5
47*
42
Y
Y
6.09
27.2
1.01
0.65
6
30
-
N
Y
0.52
0
-
-
7
30
30
Y
Y
5.45
19.6
0.54
0.65
8
106
58
Y
Y
5.41
30.6
0.41
0.53
9
71*
43.4
N
Y
7.28
35.6
0.56
0.82
10
55*
23
N
Y
4.35
17.1
0.31
0.73
RRB2, York, September 2006
Succinic acid production
Succinic acid (g/L)
40
40
No pH
control
30
30
20
20
10
10
0
1
2
3
4
5
6
7
8
Batch number
RRB2, York, September 2006
9
10
0
Can a cereal based medium replace Medium 1?
US Patent
Glucose
5,573,931
Yeast Extract
B12
Biotin
Folic acid
Riboflavin
Niacin
Pantothenate (B5)
p-aminobenzoate
Thiamine
Pyridoxine (B6)
NaCl
NaH2P4∙H2O
Na2HP4
CaCl2∙2H2O
MgCl2∙6H2O
Vitamins
Hydrolysate
Can flour hydrolysate
replace glucose?
Minerals
RRB2, York, September 2006
o se
glu
c
30
Com
me
rc
30
40
ial
No pH
control
Flo
u
Succinic acid (g/L)
40
r h
y dr
oly
sat
e
Testing the flour hydrolysate
20
20
10
0
10
1
2
3
4
5
6
7
8
Batch number
RRB2, York, September 2006
9
10
0
Can a cereal based medium replace Medium 1?
US Patent
Glucose
5,573,931
Yeast Extract
B12
Biotin
Folic acid
Riboflavin
Niacin
Pantothenate (B5)
p-aminobenzoate
Thiamine
Pyridoxine (B6)
NaCl
NaH2P4∙H2O
Na2HP4
CaCl2∙2H2O
MgCl2∙6H2O
Vitamins
Hydrolysate
Can flour hydrolysate
replace glucose?
Minerals
Can flour hydrolysate
replace vitamins?
RRB2, York, September 2006
9
Medium 1 without vitamins
Testing the flour hydrolysate
Medium 1 with vitamins
Hydrolysate without vitamins
Succinic acid (g/L)
40
40
No pH
control
30
30
20
20
10
10
0
1
2
3
4
5
6
7
8
Batch number
RRB2, York, September 2006
9
10
0
Can a cereal based medium replace Medium 1?
US Patent
Glucose
Hydrolysate
5,573,931
Yeast Extract
B12
Biotin
Folic acid
Riboflavin
Niacin
Pantothenate (B5)
p-aminobenzoate
Thiamine
Pyridoxine (B6)
NaCl
NaH2P4∙H2O
Na2HP4
CaCl2∙2H2O
MgCl2∙6H2O
Vitamins
Minerals
Hydrolysate
Can flour hydrolysate
replace glucose?
9
Can flour hydrolysate
replace vitamins?
9
Can fungal autolysate
replace Yeast Extract
and minerals?
RRB2, York, September 2006
The generic feedstock as sole nutrient
Only hydrolysate + autolysate
Succinic acid (g/L)
40
40
No pH
control
30
30
20
20
10
10
0
1
2
3
4
5
6
7
8
Batch number
RRB2, York, September 2006
9
10
0
Can a cereal based medium replace Medium 1?
US Patent
Glucose
Hydrolysate
5,573,931
Yeast Extract
Hydrolysate
NaCl
NaH2P4∙H2O
Na2HP4
CaCl2∙2H2O
MgCl2∙6H2O
Minerals
Autolysate
Autolysate
Can flour hydrolysate
replace glucose?
9
Can flour hydrolysate
replace vitamins?
9
Can fungal autolysate
replace Yeast Extract
and minerals?
9
RRB2, York, September 2006
Can a cereal based medium replace Medium 1?
US Patent
Hydrolysate
5,573,931
Autolysate
Hydrolysate
Autolysate
Can flour hydrolysate
replace glucose?
9
Can flour hydrolysate
replace vitamins?
9
Can fungal autolysate
replace Yeast Extract
and minerals?
9
RRB2, York, September 2006
Can a cereal based medium replace Medium 1?
Batch
10
Glucose
FAN
(g/L)
(mg/L)
55*
200
YE
Vitamins Minerals
Hydrolysate
Y/N
Y/N
Y/N
N
N
Autolysate
N
pH control
Y/N
Y
OD660US SA
Productivity
Patent
(g/L)
(g/L/h)
5,573,931
4.35
17.1
0.31
Hydrolysate
Autolysate
Can flour hydrolysate
replace glucose?
9
Can flour hydrolysate
replace vitamins?
9
Can fungal autolysate
replace Yeast Extract
and minerals?
9
RRB2, York, September 2006
Yield
0.73
Can a cereal based medium replace Medium 1?
Batch
10
Glucose
FAN
(g/L)
(mg/L)
55*
200
YE
Vitamins Minerals
Hydrolysate
Y/N
Y/N
Y/N
N
N
Autolysate
N
pH control
Y/N
Y
OD660US SA
Productivity
Patent
(g/L)
(g/L/h)
5,573,931
4.35
17.1
0.31
Hydrolysate
Autolysate
Can flour hydrolysate
replace glucose?
9
Can flour hydrolysate
replace vitamins?
9
Can fungal autolysate
replace Yeast Extract
and minerals?
9
RRB2, York, September 2006
Yield
0.73
Can a cereal based medium replace Medium 1?
FAN
(g/L)
(mg/L)
55*
200
YE
Vitamins Minerals
Hydrolysate
Y/N
Y/N
Y/N
N
N
Autolysate
pH control
Y/N
N
Y
Hydrolysate
Autolysate
OD660US SA
Productivity
Patent
(g/L)
(g/L/h)
5,573,931
4.35
17.1
0.31
4
15
Can flour hydrolysate
10
replace glucose?
9
Can flour hydrolysate
Succinic
acid
9
replace vitamins? OD
0
5
0
20
40
Can fungal
autolysate
(h)
replace Yeast Time
Extract
and minerals?
0.73
5
20
RRB2, York, September 2006
Yield
60
9
3
2
1
0
-1
OD660
10
Glucose
Succinic acid (g/L)
Batch
Can a cereal based medium replace Medium 1?
FAN
(g/L)
(mg/L)
55*
200
YE
Vitamins Minerals
Hydrolysate
Y/N
Y/N
Y/N
N
N
Autolysate
pH control
Y/N
N
Y
Hydrolysate
Autolysate
OD660US SA
Productivity
Patent
(g/L)
(g/L/h)
5,573,931
4.35
17.1
0.31
4
15
Can flour hydrolysate
10
replace glucose?
9
Can flour hydrolysate
Succinic
acid
9
replace vitamins? OD
0
5
0
20
40
Can fungal
autolysate
(h)
replace Yeast Time
Extract
and minerals?
0.73
5
20
RRB2, York, September 2006
Yield
60
9
3
2
1
0
-1
OD660
10
Glucose
Succinic acid (g/L)
Batch
Downstream Processing
Wheat
Upstream
processing
Bioreaction
Downstream
processing
B2
Succinic
acid
6
10
7
8
9
z Crystallisation
z Extraction by ionic liquids
z Adsorption
z Electrodialysis with bipolar membranes (EDBM)
z Reactive extraction, vacuum distillation and
crystallisation
RRB2, York, September 2006
Downstream Processing
Wheat
Upstream
processing
Bioreaction
Downstream
processing
B2
Succinic
acid
6
10
7
8
9
z Crystallisation
z Extraction by ionic liquids
z Adsorption
z Electrodialysis with bipolar membranes (EDBM)
z Reactive extraction, vacuum distillation and
crystallisation
RRB2, York, September 2006
Conclusions
z Key factors influencing bioproduction of succinic acid:
Micro-organism, pre-culture, pH control, Vitamins
z Achieved a succinic acid production of 35.6 g/L, with
a yield of 0.82 g SA/g glucose.
z It is possible to produce SA using only flour
hydrolysate and fungal autolysate from wheat grain
processing.
z Platform chemicals can be produced from renewable
resources
RRB2, York, September 2006
Acknowledgements
Miss Carol Lin
Dr Chenyu Du
Dr Apostolis Koutinas
Dr Ruohang Wang
RRB2, York, September 2006