バイオマス科学への招待

Introduction to Biomass Science

福島 和彦

Kazuhiko Fukushima

Nagoya University

Graduate School of Bioagricultural Sciences

G-COE Nov 13, 2009

Introduction to Biomass Science

1. Why do we use the biomass (lignocellulose)?

2. What is lignocellulose?

3. Structure of lignin

4. Enzymatic saccharification of lignocellulose

5. Conclusion

Topics

9.3％の排出削減が必要

京都議定書削減約束

森林吸収対策

京都メカニズム

なぜ、今バイオマス・ニッポンなのか？

バイオマス･ニッポン総合戦略より

1. Carbon stock (fixation of CO2 & use of woody material after harvest)

炭素貯蔵効果：森林による二酸化炭素吸収固定、使用中における貯蔵

2. Low energy during the process of production compared with other materials.

省エネ効果：他の材料と比較して製造時における少ないエネルギー

3. Energy substitution effect for the fossil resources

エネルギー代替効果：化石燃料の消費節約による二酸化炭素放出削減

Gakujyutu no Doko 2008 Nov

Advantages for the usage of woody biomass (lignocellulose) to reduce CO2

Introduction to Biomass Science

1. Why do we use the biomass (lignocellulose)?

2. What is lignocellulose?

3. Structure of lignin

4. Enzymatic saccharification of lignocellulose

5. Conclusion

Topics

Annual Report on Trends in Forest and Forestry 2008, Forestry Agency

When, Where, How Lignification Occurs?

Cross section

Cryptomeria japonica PH: phloem, CZ: cambial zone,

XY: xylem

S1 formation

After S3 formation

Structure of lignocellulose

Structure of cellulose

Inter cell wall

lignin

Inter fibril

Wood cell wall Aromatic ring

Hydrogen bond

glucose

Lignocellulose is very complicated structure composed by carbohydrates and phenylpropanoides which are quite different chemical natures.

Development of highly efficient saccharification of unused woody biomass

1. Why do we use the unuesed woody biomass (lignocellulose)?

2. What is lignocellulose?

3. Structure of lignin

4. Enzymatic saccharification of lignocellulose

5. Conclusion

Lignin in Tree Xylem

Plant cell wallsCellulose 50%Hemicellulose 20-30%Lignin 20-30%

HC

OCHOHC HC

CHCH2OH

OHOCH3

O

H3CO

HOHCCHCH2OH

O

CHOHCH

H2COHO

H3CO

HC O CH2CHCH2OH

HO

H3CO

HC OCHOH

OHOCH3

OCH3

HCHC

OOCH3

CHCHCHO

OHCOCH

CH2OH

OOCH3

CHOHHC O

HOH2C

CHOHC O

OCH3

HC OCHCH2OH

HO

H3CO

H3CO

CHCHCH2OH

OH3CO

CH2

CH2

CH2OH

H3CO

CH2

HC

CH2OH

OH

OCH3O

H3CO

HCCHCH3

OHH3CO

CH

Carbohydrate

CHCH2OH

CH2OH

OH3CO

CHCHCH2

OH2CHCHC

O

O

OCH3

CO

HCHOH2C

CHOH

OCH3

OH

CHCH

OHCHC

HOH2C

OHH3CO

OH3CO

HC OOCH3

HOHCHC

H2C

CHOHC CH2OH

OHOCH3

CHOCHCH2OH

OH3CO

COCHCH2OH

OH3CO

CHOHCHOH

H2C O

H3CO

H2COH

H2COH

HOH2C

Lignin polymer

Wood (lignocellulose) = Major part of biomass (90%)Key for the sustainable society

Biosynthesis of lignin and compositions of lignin structural units in various plants

Grass

GSH lignin

（monocotyledon）

Monolignol (lignin monomer)

ｐ-Hydroxy-phenyl(H) unit Guaiacyl (G) unit Syringyl (S) unit

phenylalanine

dehydropolymerization

Hardwood

GS lignin

（angiosperm）

Softwood

G lignin

（gymnosperm）

Relationship between the evolution of plant and the lignin structure

Glucose

photosynthesis

Biosynthesis of monolignol

CO2 + H2O

ｐ-coumaryl alcohol Coniferyl alcohol Sinapyl alcohol

Basic building unit

HC

OCHOHC HC

CHCH2OH

OHOCH3

O

H3CO

HOHCCHCH2OH

O

CHOHCH

H2COHO

H3CO

HC O CH2CHCH2OH

HO

H3CO

HC OCHOH

OHOCH3

OCH3

HCHC

OOCH3

CHCHCHO

OHCOCH

CH2OH

OOCH3

CHOHHC O

HOH2C

CHOHC O

OCH3

HC OCHCH2OH

HO

H3CO

H3CO

CHCHCH2OH

OH3CO

CH2

CH2

CH2OH

H3CO

CH2

HC

CH2OH

OH

OCH3O

H3CO

HCCHCH3

OHH3CO

CH

Carbohydrate

CHCH2OH

CH2OH

OH3CO

CHCHCH2

OH2CHCHC

O

O

OCH3

CO

HCHOH2C

CHOH

OCH3

OH

CHCH

OHCHC

HOH2C

OHH3CO

OH3CO

HC OOCH3

HOHCHC

H2C

CHOHC CH2OH

OHOCH3

CHOCHCH2OH

OH3CO

COCHCH2OH

OH3CO

CHOHCHOH

H2C O

H3CO

H2COH

H2COH

HOH2C

Non-repeat unitsDifficult to isolate

p-Hydroxyphenyl

OHOCH3

OH OHOCH3H3CO

H G S

Guaiacyl Syringyl

CCC

O

CCC

O4

5'

CCC

O

CCC

O5 5'

CCC

O

C8 1'

CCC

O

O

C

8 5'

CCC

O

CCC

O

8 8'

CCC

O C

O

8 4'

8-5’5-5’

4-O-5’

8-1’

8-8’

8-O-4’

Main inter-units linkages in lignin

etc.

Radical coupling polymerization

OCH3

O

CH2OH

OCH3

O

CH2OH

H

Ra Rb

OCH3

O

CH2OH

OCH3

O

CH2OH

Rc Rd

OCH3

O

CH2OH

Re

Lignin model (Sakakibara, 1980)

Coniferyl alcohol

OCH3

OH

CH2OH

- H

oxidase

Ligninmonomer

Radical mixtures

Peroxidase etc.

Heterogeneous distribution (age of cell, kind of cells)

Introduction to Biomass Science

1. Why do we use the biomass (lignocellulose)?

2. What is lignocellulose?

3. Structure of lignin

4. Enzymatic saccharification of lignocellulose

5. Conclusion

Topics

Production of Bioethanol

USA

BrazilProduction of bioethanol from sugarcane since 1970’s

Production of bioethanol is about 17 million kL/year (2005).

Japan

Production of bioethanol is 18.5 million kL/year (2006).

it is planed that the bioethanol production will be boost up to 130 million kl/year until 2017 (according to the State of the Union Address).

Some pilot plants are working (2 plants use woody materials)

ＲＩＴＥ Report

17

13000

60,000,000 kL (Japan)

1840

Prod

uctio

n/10

,000

kL

prospect

year

Background• Utilization of Biomass

Bioethanol Raw materials:

• Easily conversion to ethanol

• Food

Bioethanol VS Food

It is important to utilize the woody materials.（90% of the total terrestrial biomass ）

mainly cornstarch and sugarcane

Illustration of wood structure

Lignin

Hemicellulose

Cellulose

Bark

Group of dead cells(only cell walls)

To Molecular Level

Secondary wall

Sapwood

Heartwood

Pith

Cambium

Hinoki cypressSapwood：ray cell---livingHeartwood：ray cell---dead

To Cell Level

Fixation and stock of CO2

Oriented crystalline cellulose surrounded by lignin-hemicellulose matrix

Difficult to biodegrade

Complicated nano-material

Woody biomass

CornSaccharification bioethanol

Sugarcane

Outline of Bioethanol production system

fermentation

bioethanolPre-

treatment fermentation

Easy

Difficult

bioethanolfermentation

Saccharifi-cation

Difficult

R&D BackgroundBark is a reservoir of biomass resources because it possesses abundant starch, cell wall polysaccharides, and so on. However, it is only utilized for thermal recycling by burning or for compost and it has not been subjected to conversion to liquid fuel.

R&D Themes and Objectives

(1)Customized Aspergillus oryzae for saccharification of bark will be developed to establish revolutionary solid state enzymatic saccharification.

(2)Technology to breed trees suitable for highly-efficient saccharification (by acid or enzyme) will be developed.

(3)Through this research, sugar which can be converted to about 170 L of ethanol will be obtained from 1 t of bark.

未利用バイオマス（樹皮）の糖化（NEDOバイオマスエネルギー先導技術開発）

Effect of hydrothermal pretreatment with carbon dioxide

0

5

10

15

20

25

30

35

40M

onos

acch

arid

es y

ield

(%)

FructoseMannoseXyloseGlucoseGalactoseArabinoseFucose

150℃

1 h 2 h 4 h

175℃

1 h 4 h

200℃

1 h 2 h 4 hNo pre-treatment

WithoutCO2,

200℃, 1 hwith CO2 with CO2 with CO2

2 h

Yield (% of biomass) of monosaccharides (fructose, mannose, xylose, glucose, galactose, arabinose and fucose) by enzymatic hydrolysis after pretreatment.

Eucalyptus inner bark

0

5

10

15

20

25

30

35

40

45

50

前処理無し 175℃, 1h 175℃, 1h 175℃, 2h 175℃, 4h

糖収

率 (

%) Fru

Man

Xyl

Glc

Gal

Ara

二酸化炭素添加処理二酸化炭素無添加処理

Effect of hydrothermal pretreatment with carbon dioxide

Eucalyptus xylem

Mon

osac

char

ides

yie

ld (%

)

No pre-treatment

Yield (% of biomass) of monosaccharides (fructose, mannose, xylose, glucose, galactose, arabinose and fucose) by enzymatic hydrolysis after pretreatment.

without CO2 with CO2

SEM photograph of phloem fiber (A) of Eucalyptus inner bark in the control section. (B) after the treatment with acidified hot water with CO2 at 200°C.

(A) (B)

Introduction to Biomass Science

1. Why do we use the biomass (lignocellulose)?

2. What is lignocellulose?

3. Structure of lignin

4. Enzymatic saccharification of lignocellulose

5. Conclusion

Topics

HC

OCHOHC HC

CHCH2OH

OHOCH3

O

H3CO

HOHCCHCH2OH

O

CHOHCH

H2COHO

H3CO

HC O CH2CHCH2OH

HO

H3CO

HC OCHOH

OHOCH3

OCH3

HCHC

OOCH3

CHCHCHO

OHCOCH

CH2OH

OOCH3

CHOHHC O

HOH2C

CHOHC O

OCH3

HC OCHCH2OH

HO

H3CO

H3CO

CHCHCH2OH

OH3CO

CH2

CH2

CH2OH

H3CO

CH2

HC

CH2OH

OH

OCH3O

H3CO

HCCHCH3

OHH3CO

CH

Carbohydrate

CHCH2OH

CH2OH

OH3CO

CHCHCH2

OH2CHCHC

O

O

OCH3

CO

HCHOH2C

CHOH

OCH3

OH

CHCH

OHCHC

HOH2C

OHH3CO

OH3CO

HC OOCH3

HOHCHC

H2C

CHOHC CH2OH

OHOCH3

CHOCHCH2OH

OH3CO

COCHCH2OH

OH3CO

CHOHCHOH

H2C O

H3CO

H2COH

H2COH

HOH2C

filtrate

residue

fermentation

Extract of ligninprecursors

Bioethanol

Law materials for

green chemistry

Lignin

Effective separation of cellulose from lignocellulose

Pretreatment

Saccharification

BioinformaticsFast growing treeRegulation of lignin metabolic map

Extract of phenolicsfrom tissue culture

HO

OCH3

CO2H

O

CH2OHH3CO

glucose

WoodTree

Tree biothecnology

Bioplastics Partialmodification

Biomimetic polymerization（usage as a monomer）

Medicines

Biorefinery based on lignocellulose

ＣＯ２

ＣＯ２

Substitute for petroleum

Carbonneutral

なぜ、今バイオマス・ニッポンなのか？

Annual Report on Trends in Forest and Forestry 2008, Forestry Agency

収入

整理･統合

林道整備・機械化