Usos de la fermentación en la producción masiva

de compuestos químicos a partir de biomasa

Bioeconomía Argentina 2013: Biomasa, innovación y valor agregado, Buenos Aires, March 21-22, 2013

Ruud A. Weusthuis

How can we use fermentation for the

production of bulk chemicals?

Argentinian Bioeconomy 2013: Biomass, innovation and value added; Buenos Aires, March 21-22, 2013

Ruud A. Weusthuis

Why fermentation processes?

Example: glucose to ethanol

12 reactions

Chemistry:

● 90% efficiency per step, 28% overall

● Multiple reactors, high investment costs

Fermentation

● 90% efficiency overall

● One pot synthesis, low investment costs

Market prices

Compounds Price €/ton

Petrochemicals Polyethylene 700

Acetic acid 400

Ethylene glycol 900

Fermentation products

Ethanol 600-700

Lactic acid 1800-2250

Citric acid 800

L-glutamic acid 1200

L-Lysine 2000

Itaconic acid 4000

Making bulk chemicals with

microorganisms

Product Substrate Fermen-tation

Product recovery

Goal: competitive with fossil derived products

Yield (substrate efficiency), productivity, titre

Substrate

Product

Which type of product?

0

10

20

30

40

50

60

70

80P

rod

ucti

on

costs

€/

GJ e

nd

prod

uct

Capital

Oil/gas

Coal

Added value: 2 - 5 times higher

Which type of substrate?

Side streams, lignocellulose

Which type of substrate?

Side streams, lignocellulose

Complex:

● Implications for product recovery

● Fermentation inhibitors present

● Mixture of carbon sources

Low sugar concentrations (~50 g/l)

Mixing problems

● 2-3 times lower oxygen transfer

Substrate costs: 2 x as low?

What type of fermentation?

Ethanol: 0.95 J/J

Lactate: 0.95 g/g

L-glutamic acid: 0.62 g/g

Itaconic acid: 0.47 g/g

Anaerobic: Aerobic:

Anaerobic fermentations

Aerobic Anaerobic

Productivity Limited by O2 transfer No O2 required

Limited by cooling capacity Low heat production

Substrate efficiency

High biomass production Low biomass production

Complete oxidation of substrate to CO2

No complete oxidation

Anaerobic fermentations: challenges

CO2-fixating pathways

Glucose

2 Pyruvate

Electrons

2 Lactate

Electrons

Glucose

2 Pyruvate

2 CO2

Succinate

Electrons

Electrons

O2

Water

Glucose

2 Pyruvate

2 CO2

Succinate

Electrons

Electrons

Glucose

Succinate

Protons

Hydrogen gas

Hydrogen producing pathways

Aerobic Anaerobic

3-hydroxybutyric acid

Monomer of the polyester PHB

PHB competes with polyethylene

Anaerobic production:

2 glucose 3-hydroxybutyric acid + 2 ethanol + 4 H2 + 4 CO2

Anaerobic fermentations

Yield: 0.95 g/g or J/J

Productivity: up to 5 times higher

4 projects running

Product inhibition

0

10

20

30

40

50

60

70

80

90

0 50 100 150 200

[P

rod

uct]

% o

f h

igh

est

con

cen

trati

on

Time (% total time)

Productivity, titre and yield are limited by product inhibition

25 50 75 100

Prevent product inhibition

Strains with improved product tolerance

● Product tolerance difficult to engineer

● Isolate strains with high tolerance

● Convert them into producers of chemicals

● Example: Isolated strain Monascus ruber LF6 is

able to grow in presence of 150 g/l lactic acid at pH

2-3 (WO2012055996-8)

3 projects running

Apply in-situ product recovery

Examples:

Ethanol production at low pressure

● 4 g/l/h at atmospheric pressure

● 40 g/l/h at low pressure

● Cysewski and Wilke, 1977

Lactic acid in continuous membrane reactor

● 4-7 g/l/h in normal batch

● 22 g/l/h in CMR

● Tejayadi and Cheryan, 1994

Prevent product inhibition

Productivity: up to 5-10 times higher Titre: up to 5 times higer

2 projects running

Product recovery

High costs of product separation:

Up to 20-40% of total costs

Biofuels in fermentation broth

Ethanol: Miscible

Distillation, 20% of energy

Dodecanol (C12): Immiscible

Simple separation!

Example: LS9, Amyris

Phase separations

Liquid/liquid

● Making water immiscible compounds

Gas/liquid

● Volatile compounds, high temperature

fermentations

Solid/liquid

● Making polymers: Cyanophycin,

polyhydroxyalkanoates

● Crystals

No/less product inhibition!

Phase separations

2-4 times as cost effective?

2 projects running

Recipe for application of metabolic

engineering for production of bulk products

1. Make chemicals, not fuels: 2-5 x more added value

2. Use inexpensive side streams: 2 x lower substrate costs?

3. Use anaerobic fermentations: Yield to 0.95 g/g, J/J

Productivity: 5 x as high

4. Prevent product inhibition: Productivity: 5 x as high

5. Use phase separation: 2-4 x as cost effective?

Take-home message:

Production costs of fermentation

products can be reduced below

that of bioethanol

Thanks to

Gerrit Eggink

Leo de Graaff

Annemarie Hage

Serve Kengen

Ischa Lamot

Audrey Leprince

Mark Levisson

Astrid Mars

Odette Mendes

EOS-NEO program

BE-BASIC program

Bas Meussen

Youri van Nulandt

John van der Oost

Mark Roghair

Johan Sanders

Peter Schaap

Jan Springer

Kiira Vuoristo

Hetty van der Wal

Emil Wolbert

End

Thank you for your attention

Biobased Commodity Chemicals

Ruud Weusthuis

Fermentation

processes

Johan Sanders

Biorefinery

Marieke Bruins

Biobased Process

Technology

Elinor Scott (Bio)Catalytic

Transformations