Biorefinery in Germany –

Current Status

Thomas Willke
Federal Agricultural Research Centre
Institute of Technology and Biosystems Engineering
Bundesallee 50, D-38116 Braunschweig
Email: thomas.willke@fal.de - telephone: ++49 (0)531-596-4124

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Outline
Î Processes, Feedstocks
¾ Sugar-refinery, starch/sugar
¾ Oil-mill/biodiesel plant, oil/fats
¾ Lignocellulose, wood, straw,…
Î Technologies
¾ Cell-/substrate-screening,
genetic Engineering
¾ Fermentation (aerobic,
anaerobic)
¾ Chemical catalysis (Sugar,
Fermentation, Products)
¾ Immobilisation Techniques
Î Examples
¾ Itaconic acid (sugar, oil?)
¾ Ethanol (sugar)
¾ Palatinit (sugar)
¾ Biotensides (sugar + oil)
¾ 1,3-Propanediol (oil)
¾ Catalytic sugar oxidation
(sugar)
¾ Biogas (residues,all)
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Processes
Starch, Sugar

Sugar-Refinery (for example: Südzucker, crop energies)

Starch-/ Sugar Plants Sucrose Ethanol

Residues Glucose Products

Biogas
Glucose/Fructose Products

Electricity,
Heat Fructose Products

Palatinit

Fertiliser, Animal feed

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Technical polymers
examples

polymer group name monomer use

polyamide Nylon®, Perlon® diamines, fabrics, carpets

Kevlar® dicarbonic acids foils high-performance fibers,
cars

polycarbonate Makrolon® dioles CDs, flasks, synthetic glasses

polyester Trevira®, Sorona®, dioles, dicarbonic flasks, fabrics

Corterra® acids carpets
high-performance fibers

polyurethane polyoles foams, resins, glues, lacquers

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Polymers: Starting Material
Acids (accessible from renewable resources)
COOH
fumaric
itaconic
HOOC O
HOOC HOOC COOH
2-keto-glutaric
OH COOH
HOOC
citric succinic
COOH COOH
HOOC COOH

HO COOH HOOC aconitic

3-HP (3-hydroxy propionic)
COOH COOH
HO 2-HP (lactic) COOH
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The Tricarbonic-Acid-Cycle
Potential substrates und products
Starch glycolysis
sugar ACETYL- CoA
Fats/oil
glycerol
Oxal-acetic
Oxalessigsäure
citric
Citronensäure
O COOH
COOH
COOH
malic
Äpfelsäure HO COOH

HO COOH COOH

HOOC COOH COOH

COOH COOH

COOH COOH

HOOC
cis-Aconitsäure
cis-aconitic itaconic
Itaconsäure

fumaric
Fumarsäure
COOH
COOH
COOH
HO COOH
COOH COOH Isocitronensäure
Iso-citric
Bernsteinsäure
succinic CH2
O COOH

α-Ketoglutarsäure
ketoglutaric
(2-Keto-glutarsäure) TW-020117
Itaconic Acid

sugar, starch
(glucose) complexing agent
Aspergillus terreus
(fungi)

co-monomer,
plasticiser

itaconic acid
paints, lacquers
Yarrowia spp., Candida spp.
(yeast)

fats, oil adhesives

glycerol

others

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 Itaconic Acid Production
Fermentation from glucose

200 100
conditions
180 90 „ batch
(without pH-
160 itaconic acid 80
controll)
140 70 „ mineral-salt

Itaconic acid [g/L]

medium,
Glucose [g/L]

120 60
P-limitation
100 50 „ pH 1,7
„ 33°C
80 40

60 30
result
40 20 Î final concentration
glucose > 80 g/L IA
20 10
Î max. productivity
0 0 > 1 g/(L·h)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Î Yield: 0,6 g/g =
time course[d] 83% of Theory

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Itaconic Acid Production
other substrates?

80

70 Glucose
Saccharose
60
Itaconic acid [g/L]

50
Glycerol
40

30

20

10
Lactose
0
0 2 4 6 8 10 12 14 16 18 20 22 24
time [d]

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Immobilisation in Biotechnology
Example: ethanol

biofuel,
chemical feedstock
bead surface

yeast-cells
wheat

x 100

sugarbeet
HO CH2
O HO CH2
HO
O
O HOH2C O HO HO
OH CH2OH

OH
O
HO OH
O
O
OH H O CH2 CH3
HOH2C O H2C O HOH2C O

O
immobilised
O
HO OH
O
HO OH HO OH O

yeasts or bacteria ethanol

starch, sucrose (biokatalyst)
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Immobilisation in Biotechnology
Types & shapes of gel

cube particle
foil

fiber hollow fibre

single cell

lens
hollow bead bead top view

cross section

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Bead Production with JetCutter
Scheme and operation
pressure

cutting nozzle
tool
motor D

cutting n ufluid liquid

M jet
wire
or uwire

cut cylinder

bead

ufluid = fluid velocity m = motor D = diameter of

uwire = wire velocity n = rotations the nozzle
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JetCutter
Details of the cutting tool

cutting tool equipped with

23 µm PET fibres

cutting tool with circular

stabilization of the wires
(48 wires, diameter: 0.1 mm)

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JetCutter
optimisation of settings for low spray losses

suited adjustments
Æ almost no losses

unsuited adjustments
Æ high losses

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JetCutter
Particle size distribution

-1] -1
[mmmm 14

12
Ca-Alginat
density,

10
distribution density

6
distribution

PVA
4

0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2
beaddiameter
bead diameter, mm
[mm]
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JetCutter
Examples of hydrogel beads

Ca-alginate Ca-pectinate
Ø = 300 µm Ø = 500 µm

gelatin chitosan
Ø = 800 µm Ø = 500 µm

not in true scale!

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JetCutter
Equipment

lab scale

production scale
pilot scale capacity: up to 1.2 t/d (Ø 1 mm)
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Bioethanol Production
Yeast or bacteria?

free cells immobilized cells

microorganism activity activity productivity

[kg EtOH/(kg BDM·h)] [kg EtOH/(m3 cat·h)] [kg EtOH/(m3·h)]

yeast
Saccharomyces 0.5 – 1.4 25 - 70 10 - 30
cerevisiae

bacterium
4-5 190 - 230 50 - 80
Zymomonas mobilis

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Ethanol Fermentation: Production Plant
Conventional or with immobilised cells
conventional fermentation biomass
recycling
separator

nutrients
6 x 200 m3
molasses
ethanol

buffer

Separator
(smaller)
process design with
ethanol
immobilised cells
buffer
nutrients
molasses 3 x 60 m3
BMA Braunschweig, capacity: 60,000 litres EtOH / 24 h
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Ethanol-Fermentation: Pilot Plant
Immobilised yeast, continously, 3-step

© BMA-Braunschweig

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Palatinose-Production
Example for immobilisation on an industrial scale

immobilised cells
of
Protaminobacter rubrum

saccharose palatinose

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Palatinose-Production
Bioconversion

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Industrial Production of Palatinose with
Immobilised Microorganisms

80,000 tons per year

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Production of Sugar Acids
Chemical catalysis with gold catalysts
sugar
milk T = 40-60°C
pH = 7-9

Sugar acids
Complexing agents
starch Ingredients for cosmetics,
food, pharmaceuticals
wood oxygen HO O Functional Food
C
H C OH
HO C H
Galactose C HO
Galactonsäure
H C OH HO C H
CH 2OH
O OH CH2OH
H
HO C H H C OH H OH
CH2 OH OH H
O O H CH2OH OH H COOH
OH CH2 OH
OH H H OH HO C H OH O O
COOH
H OH H H OH

H OH H C OH H H H C OH

H O
Lactose supported H OH HO C H
C CH 2 OH Lactobionsäure H C OH
H
HO
C
C
OH
H
gold catalyst H C OH
H

H
C

C
OH

OH
Glucose Gluconsäure CH2OH

CH2OH
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Oxidation of Monosaccharides
Selectivity of the gold catalyst (HPLC-data)
2,5.106 106

2.106
Peakfläche im UV-Detektor

HO O
HO O C

Signal im UV-Detektor
C H C OH
1,5.106 H C OH Gluconic Galactonic HO C H
HO C H 5.105 HO C H
6 H C OH
10 H C OH
H C OH
CH2 OH
CH2 OH
5.105

0
18 20 22 24 26 28 30 0 2 4 6 8
Zeit, min
Zeit, min

3.105 5.105

4.105
Signal im UV-Detektor

Signal im UV-Detektor
HO O
2.105 C
HO O
Xylonic 3.105
H C OH C
HO C H H C OH
Ribonic
H C OH 2.105 H C OH
105
CH2 OH H C OH

105 CH2 OH

0 0
0 2 4 6 8 0 2 4 6 8
Zeit, min Zeit, min
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Oxidation of Disaccharides
Selectivity of the gold catalyst
35 35

30 30
Lactobionic Maltobionic
Peakhöhe UV [mAbs]

Peakhöhe UV [mAbs]
25 25

20 CH2OH 20
H OH
CH2OH CH2OH
15 CH2OH OH H COOH 15
H O H H OH
OH O O
OH H OH H COOH
10 H OH 10 O
OH H OH
H H
5 5 H OH H OH
H OH

0 0
18 20 22 24 26 28 30 18 20 22 24 26 28 30
Zeit [min] Zeit [min]

6.105

Cellobionic
Signal im UV-Detektor

4.105
CH2OH
H OH

CH2OH OH H COOH
.
2 10 5 H O O

OH H H OH
OH H
H OH

0
0 2 4 6 8
Zeit, min TW-020117
Processes
Oil mills, Biodiesel Pants

MeOH EtOH

Oils Plants RME (REE), ?

Biodiesel
Residues biological
1,3-Propanediol
Glycerol
Biogas Acroleine
chemical 1,2-PD + 1,3-PD
Sugar
Electricity,
Heat
Fatty acids, fats Biotensides

Rapeseed meal Proteins, Animal Feed

Rapeseed

Fertiliser

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Biodiesel from Rapeseed
Production

RME-production
Rapeseed

seed oil 90 % biodiesel

(40...45 %)

10 % glycerol
oil by-products
rapeseed cake (6...12 % Öl)
(55...60 %)
oil mill
rapeseed meal (1...2 % oil)
about 40 % protein TW-020117
Biodiesel
Production capacity in Germany

Tons per year

2.900.000 *
3.000.000

2.500.000
1.900.000
2.000.000

1.500.000 1.200.000
1.100.000

1.000.000 730.000

460.000
500.000 140.000 200.000
50.000
0
1998 1999 2000 2001 2002 2003 2004 2005 2006

Data: UFOP, *estimated

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 price in bulk CFR Rotterdam [€/t]

0
200
400
600
800
1000
12.1998

06.1999

12.1999

06.2000

12.2000

06.2001
Price of Raw Glycerol

12.2001

06.2002

12.2002
glycerine 80% crude

06.2003

12.2003

06.2004

12.2004

06.2005
?

12.2005
2007: < 100 €/t

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Polymers: Starting Materials
Polyols (accessible from renewable resources)

1,3-propanediol

HO OH
2,3-butanediol

1,4-butanediol
HO OH HO
OH

HO OH
OH glycerol

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1,3-Propandiol from Glycerol

1,3-propanediol

glycerol 10%
(byproduct of
biodiesel production)

bacteria, anaerobic chemical feedstock

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Possible Products of 1,3-Propanediol
Applications...

Î co-monomers in PTT (=
polytrimethyleneterephthalate)
¾ base for carpets (Corterra®)
¾ special-textile fibers (Sorona®)

Î co-monomer in polyesters
¾ binders, adhesives and sealants in industry and
housebuilding, lacquers, casting resins

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1,3-Propanediol Fermentation
Strain NRRL1024 (from culture collection)

100
conditions
concentration [g/L]

„ fed-batch
80 1,3-propanediol
(pH-controlled)
„ mineral-salt
medium + YE
60 „ pH 7.0
glycerol
„ 35°C

40
results
n-butyrate Î final concentration
20 > 72 g/L 1,3-PD
Î productivity:
acetate > 1.7 gPD/(L·h)
0
0 10 20 30 40 50 60 70
time course [h]
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1,3-Propanediol Fermentation
Strain IK123 (from screening)

100
1,3-propanediol conditions
concentration [g/L]

„ fed-batch
80 (pH-controlled)
„ mineral-salt
medium + YE
60 „ pH 7.2
glycerol „ 32°C

40
result
n-butyrate Î final concentration
20 > 100 g/L 1.3-PD
Î productivity:
acetate
> 2 gPD/(L·h)
0
0 10 20 30 40 50 60 70
time course[h]
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1,3-Propandiol-Fermentation: Costs
Influence of nutrients

substitution of YE by PNC (0 €/t)

2500
€ je t
1,3-PD
2000 ROI (20 %)
depreciation
allocated fixed costs
1500
salts, NaOH direkt fixed costs
yeast extract energy
feedstock/media
1000

500
salts, NaOH
glycerol yeast extract salts, NaOH
glycerol glycerol
0
pharma- glycerol- PNC
glycerol water instead of
yeast extract

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Production of Hydroxyfatty acids
1. Fermentation of the glycolipid

Sophoroselipid

Glucose
+
Rapseed oil
Candida
bombicola

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Production of hydroxyfatty acids
1. Fermentation of the glycolipid

Sophoroselipid

time (h)

Candida bombicola

[Rau, Lang et al., 2001]

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Production of hydroxyfatty acids
2. Hydrolysis

Sophoroselipid

Sophoroselipid
300 g/L

Hydrolysis Glucose

17-Hydroxy-oleic
acid Hydroxyfatty acid
~ 110 g/L

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Processes
Lignocellulose

Pretreatment

Lignocellulose
? Glucose Products

Xylose Products

Lignin Products

? Residues,
wastewater

? Biogas

Electricity,
Heat
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Energy from Renewable Resources
Biogas
gas boiler
feedstock
crops heat

biogas sulfur removal motor-CHP

reforming electricity, heat

manure fuel cell

reforming
compression electricity, heat
biogas plant
residues
by-products liquid storage
food- and
agro-industry, e.g. fats
fuel
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Biogas Plants in Germany
number 55 PJ = 2,3 GWh Gaspower
4000
= 0,85 GWh electric power + heat +
3600 auxiliary power + losses 3400*
3200

2800 2700
2500
2400
2000
2000 1800
1650
1600

1200 1050
850
800 617
370 450
400 186
274
100 120 139 159
0
1990 1992 1994 1996 1998 2000 2002 2004 2006
year * estimated
Source: German Biogas Association and own Data
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Electricity from Biogas
fuel-cell pilot plant located in the FAL

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White Biotechnology in Germany
Companies in Germany 1 (selection)

Î Südzucker: Sugar, Palatinose, Ethanol

Î Degussa: Amino acids
Î Dow: Plastics, Monomers, Specialities
Î BASF: Vitamins, Specialities
Î Cognis: Tensides, Plasticiser, Emulsifiers ...
Î Wacker: Cyclodextrines, Fine Chemicals

Î Rütgers: Aromatics, Downstream-Technologies

Î Symrise: Flavor, Cosmetics, Food-additives

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White Biotechnology in Germany
Companies in Germany 2 (Selection)

Î B.R.A.I.N.: Enzymes, Drugs, Bioactives

Î GeniaLab: Immobilisation Technologies, Starter Cultures, Services

Î Lipoid: Health& Nutrition, Phospholipids

Î Codexis/Jülich Fine Chemicals: Enzymes, Chiral synthesis

Î Direvo: Industrial Enzymes, Biopharmaeuticals

Î BioSpring: Enzymes, Nucleic acids

Î Dr. Rieks GmbH: Cosmetics, Food-additives

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Products (> 100 Mio €)
by Fermentation

Î Amino acids: Lysin, Glutamate… (Degussa)

Î Specialities (BASF, Henkel)

Î Palatinit (Südzucker)

Î Riboflavine (Vit B2) (DSM/Roche, BASF)

Î Cyclodextrines (Wacker)

Î Organic acids: citric, gluconic… (Jungbunzlauer)

Î Ethanol (Südzucker/crop energies, Verbio et al.)

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Activities in Germany
Research & Projects

Research (selected)
Î Linke, ATB-Bornim/Potsdam: Lactic acid, PLA (Pilot Plat)
Î Steinbüchel, Münster: Biopolymere, Polyaminosäuren
Î Bornscheuer, Greifswald: Directed Evolution of Industrial Enzymes
Î Peter, Potsdam: Chitin / Chitosan
Î Lang, Rau, Braunschweig: Biotenside

FNR-Projects (running)
Î Schwarz, München/Streit, Uni-HH/Liebl, Uni-Göttingen: Enzym. Degrad. of Lignocellulose,
Nanotechnology
Î Marc, Creavis/Degussa, Homoserin + Isobutyric Acid>Methylmethacrylate
Î Scherzer, BASF/Müller, Fraunhofer-Berlin/Kleinert, Siemens: Tuning of Biopolymers
Î Fraunhofer: Cold-Swelling gels, Functionalised Starch, Materials from Modified
Polysaccharides
Î Fraunhofer-Umsicht: C4-bases Special Polymers
Î Metzger, Oldenburg: Materials from Oils and Fats
Î Schrader, Dechema/Schmidt, Stuttgart. Enz. Lactone-Syntheses, Hydroxylation of Fatty Acids,
Î Drepper, Düsseldorf/Jülich: Lipids from Renewable Resources, Special-oils and Fatty Acids
Î In Pipeline: Lignocellulose

Further information: http:\www.fnr.de

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 That‘s it...!

Thanks for your kind attention!

Open questions...?
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