The Challenge we face

https://lnkd.in/dfwUbAY
 Scientific challenges
• Improving crop varieties – yield, efficiency, land use

• Process technology, infrastructure management &

engineering, inc. energy efficiency
• New microbial strains, increased ethanol yields,
combined saccharification & fermentation

• Better enzymes

• Exploiting other agricultural sources, combining with

cellulosics
• Identifying additional product streams
Alternative technologies for lignin breakdown
 Overview
• Lignocellulose - revision

• Lignin structure & potential

• Lignin extraction

• Lignocellulosic Biorefinery

• Microbial breakdown & low input

biorefining
 Revision lignocellulose

Evans & Hedger 2008

Fungi in Bioremediation
• Bulky

• Dense

• Composite

• Annual terrestrial production

- 100 gigatonnes plant cell
wall material
 Cell wall content of different feedstocks
Source Lignin Hemicellulose* Cellulose
Sugarcane bagasse 23 28 (22) 39
Corn stover 19 28 (22) 38
Wheat straw 17 25 (19) 33
Monterey pine 26 23 (6) 42
Hybrid popular 25 21 (13) 39
Eucalyptus 27 17 (10) 48
Switchgrass 17 26 (21) 31

Values are percentage of total mass. * Brackets = % xylan Carroll & Somerville 2009

• Lignin – second most abundant polymer?

- difficult to breakdown
 Lignocellulose the challenge and opportunity
Cellulose: 33% = 231 tonnes dry weight
Hemicellulose: 25% = 175 tonnes d.w.
700 tonnes Lignin: 17% = 119 tonnes d.w.

• Estimate 10% conversion

23.1 tonnes cellulose to ethanol £8016*
17.5 tonnes hemicellulose to ethanol £6073
11.9 tonnes lignin to vanillin £110,546**
Walton Mayer & Narbad 2003 Phytochemistry, 63, 505-515; vanillin $15/Kg**
Black & Veatch: Lignocellulosic ethanol plant in the UK feasibility study, final report (NNFCC 08-007) 2008 ethanol
value £347/tonne*
Lignin structure

Guaiacyl Guaiacylsyringyl Graminaceous

coniferyl alcohol coniferyl and p-coumaryl alcohol
sinapyl alcohol
Soft wood Hard wood Grasses
 Potential compounds
Structures found in lignin:
pinoresinol biaryl ether
-aryl ether phenylcoumarane diaryl propane
biphenyl
OMe
RO OMe OH OMe
HO OH
OMe OH O
O OH OMe
O
O O
OMe OH
O OMe
OH MeO RO OAr
OMe OMe
MeO OR OH OMe

OH CH 2OH OH
OH
MeO CO 2H
HO O CHO HO OH
O OMe
MeO HO OMe
HO
OMe OH
OMe O OMe
O
OH HO 2C OMe OH

-aryl ether (51%) biphenyl (11%) OMe

diarylpropane (2%)
OMe OH
OH
pinoresinol (2%)
phenylcoumarane (12%)

• Lignosulplates (plasticisers in concrete)

 Lignin extraction
• Kraft & Sulphite pulping
 Milling
 Cooking - ~150 C, NaOH & Na2S
 Recovery
 Blowing – steam release can provide heating
 Screening & Processing
• 7 tonnes Black liquor per tonne of pulp
 Incentives to burn as fuels (mixed with diesel)
 Threatens renewable chemical feedstock industries
 Lignin availability

https://www.netl.doe.gov/research/Coal/energy-systems/gasification/gasifipedia/blackliquor
 Lignin extraction from cellulosic biofuels
• Milled wood lignin - Wood or agricultural products

 Milling – determine amount lignin solubilised

 Extract with organic solvent (e.g. dioxane)

 25-50% available lignin

 Treat will cellulases (cellulolytic enzyme lignin)

• Other methods, e.g. Anaerobic digestion

Organsolv pre-treatment of lignocellulose
 The Modern Lignocellulosic Biorefinery
• Target multiple product
streams
• Add most value – product &
process
• Challenge food production….

• Or provide stability to market

fluctuations?

Could lignin be a major target for a biorefinery in the

future?
Lignocellulosic Biorefinery - Considerations

• At what scale should the

refinery operate?

• What processes should it

carry out?

• Where would you put them?

• Who will benefit?

USDA
Could a low input localised scale method for
converting biomass to liquid be feasible?
• On farm or farm co-operative
 Overcomes storage and transportation issues

 Farmers become stakeholders in the process

• Extraction methods have to

be simpler with lower input

• Sustainable – economic &

environmentally
 How to unlock Lignocellulose?
• Traditional pulping – localised and low input?

• Super critical steam developments – H2 and glucose

• Microbial breakdown – efficient?

 Microbial breakdown of lignocellulose
White rot
Lignin and cellulose degraded
Preferentially attack lignin

Brown rot
Efficient cellulose attack
Lignin mostly intact, but modified

Soft rot
Appearance similar to brown rot
Bacteria
Attacks cellulose-rich
Less well studied layers
May act synergistically
 Fungi verses bacteria

Fungi (filamentous) Bacteria

Region of substrate utilisation

substrate substrate

Bacteria grow faster, easy to handle & inoculate, used in

aerobic & anaerobic digestion, need of N addition

Fungi penetrate the substrate, produce large range of

enzymes which are secreted ahead of growth
 The Mushroom Biorefinery
• Composts thousands of tonnes
of straw & animal waste
• Produces high value crop

• Also produces spent mushroom

compost and ‘goody water’
Bioconversion Concept
MICROBIAL
BIOREFINERY

Fatty Acids &

Oils

Sugars

Aromatics
 Extraction methods
• Aqueous extraction
 Hot water short time, cold long time

 Sugars & water soluble aromatics (e.g. vanillin)

• Solvent extraction – room temperature?

 Variable solvent – balance target with cost & safety
e.g. hexane – fatty acids
Ethanol, dichloromethane, tolulene, ethyl ether etc..
- aromatics
 Product release from straw using fungi
Aq.soluble Solvent soluble
Treatment Sugars aromatics aromatics

Agaricus bisporus 5.58 0.36 9.5

Lentinula edodes 4.07 0.43 14.3

Lepista nuda 5.27 0.41 14

Phanerochaete chrysosporium 14.53 1.41 25.5

Pleurotus ostreatus 3.48 0.35 17

Schizophyllum commune 7.91 3.33 15.8

Serpula lacrymans 8.43 2.79 32.3

Trametes versicolor 6.68 0.92 15.8

Uninoculated control 2.64 0.75 13.3

mg dry wt g-1
Microbial successions used to target different
parts of lignocellulose
Sugars released after a first fer- Aromatics released after a first
mentation with white rot fungi and fermentation with white rot fungi
a second fermentation with brown and a second fermentation with
rot fungi brown rot fungi
0.5
0.8 0.9

0.7 0.8
0.4
0.6 0.7
S Commune 0.6 S Commune
0.5 S Commune S Commune
0.3

Phenols (%)
Sugars (%)

0.5
0.4
0.4
0.3 0.2
0.3
0.2
0.2
0.1
0.1

0.0 S commune P chrysosporium

S commune P chrysosporium -0.1
Species of the first fermentation Species of the first fermentation
Before Week 1 Week 2 Week 3

Before Week 1 Week 2 Week 3

 Comparison: single fermentations (brown rot fungi) &
sequential fermentations (brown rot & white rot fungi) LC-MS
Solvent soluble aromatics ~ 1.4 – 3.5 % dry wt conversion of straw
P. placenta S. lacrymans

P. placenta followed by S. lacrymans followed by

P. chrysosporium P. chrysosporium
 Fatty Acid Production
ORGANISM SUBSTRATE Fatty acids

Pleurotus straw 16:0, 18:2

ostreatus

Pleurotus straw + nitrogen 16:0, 18:1

ostreatus 18:2, 20:3
Serpula straw 16:0, 18:2, 20:3,
lacrymans 22:0

Trametes straw 16:0, 18:1

versicolour 18:2, 20:3
22:0

Phanerochaete straw 16:0, 18:1

chrysosporium 18:2, 20:3

Schizophillum commune straw 16:0, 18:1

18:2, 20:3
 Omega-3 Fatty Acids
• Essential dietary fatty acids

• Historically from fish, but cannot meet future demands

• Some microalgae produce at ~£40 per g

• Reduced costs if produced as part of a

multiple product stream?
 Dispersed pre-treatment model

www.themushroombusiness.com

Soluble carbohydrates
Phenolic residues from
(sugars) from cellulose
lignin breakdown & fatty
and hemicellulose
acids

Processed in
centralised facility