Hydrothermal pretreatment of biomass for ethanol fermentation · 1 Biomass Project Research Center,...
Transcript of Hydrothermal pretreatment of biomass for ethanol fermentation · 1 Biomass Project Research Center,...
Biomass Project Research Center, Hiroshima Univ. 1
Hydrothermal pretreatment of biomass
for ethanol fermentation
Dec. 10-12, 2012
JAPANESE-DANISH JOINT WORKSHOP “Future Green Technology”
Hakata, Japan
Yukihiko Matsumura
Hiroshima University
Biomass Project Research Center, Hiroshima Univ. 2
First generation Second generation
•Sugar, Starch, grain
•Easy fermentation to bioethanol
•High price raw material
•Competition with foods
•Lignocellulosic residues (wood,
straw) and other agricultural
residues
•Advanced technology is needed
緒言 First and second generation ethanol
Biomass Project Research Center, Hiroshima Univ. 3 3
Enzymatic
hydrolysis Hemicellulose
Lignin
Lignocellulosics
Ethanol
Ethanol
fermentation
CH2OH
HOH
OH
OHOH
HH
H
H
Glucose
O
OH
OH
HO
CH2OH
O
O
OH
OH
CH2OH
O
OH
OH
CH2OH
O OH
n-2
Cellulose
緒言 Saccharification of lignocellulosics
Pretreatment
Biomass Project Research Center, Hiroshima Univ. 4
Pretreatment Concept Disadvantage Author (year)
Previous study
Concentrated
sulfuric acid
Promote hydrolysis with
concentrated sulfuric acid
・Decomposition of glucose
by acid
・ High cost to use acids
Gupta R et al.
(2009)
Dilute sulfuric
acid
Promote hydrolysis with
dilute sulfuric acid
・High cost to treat
byproducts
・Reactor corrosion
Root et al. (1959)
Steam explosion After heating up in steam,
suddenly reduce the
pressure
・Low glucose yield DeLong (1981)
Pulverization Decrease the crystallinity of
cellulose
・Large amount of energy
needed.
Sidiras and Koukios
(1989)
Hydrothermal Hemicellulose is dissolved
in water by high
temperature and pressure.
Reduction of crystallinity of
cellulose.
・Low cost
・Low glucose yield
Mok and Antal
(1994)
Various pretreatment for saccharification
Biomass Project Research Center, Hiroshima Univ. 5
Hydrothermal
Pretreatment
Enzymatic
hydrolysis Ethanol
fermentation
Lignocellulosic
biomass Cellulosic
component
Monosaccharide
Ethanol
Inhibitors
Inhibitor byproducts for fermentation
Biomass Project Research Center, Hiroshima Univ. 6
Fermentation inhibitors are produced during
hydrothermal pretreatment, which affects the activities
of the yeast
Inhibitors Formic acid, Acetic acid, Furfural , 5-HMF
Yeast Saccharomyces cerevisiae
Anaerobic
Aerobic
Ethanol fermentation
Cell growth
Fermentation inhibitors
Biomass Project Research Center, Hiroshima Univ. 7
This evaluation has not been reported so far.
To commercialize the process, reaction characteristics
as well as inhibitor effect should be clarified.
but・・・
The purpose of this study is to determine the
reaction characteristics and inhibitor effect
quantitatively.
Purpose of this study
Purpose
Biomass Project Research Center, Hiroshima Univ. 8
Raw materialRubber wood residues
Hydrothermal pretreatment
Filtration
Liquid fraction Solid fraction
Glucose product
Glucose productEnzymatic hydrolysis
Autoclave reactor
The working volume of the pretreatment
vessel was 96 mL. The pretreatment
agitator was set at 500 rpm. Reactant 1 g
Buffer fluid 60 mL
10 g/L cellulase solution 5 mL
Enzymatic hydrolysis
Cellulase from Aspergillus niger powder,
≥0.3 units/mg solid
The flasks were shaken at 250 rpm at 37 C
HPLC with SUGAR K S-802(Shodex)
column operated at 60 °C with
0.8mL/min flow of water as an eluate. The
detector was a refractive index
Experimental for hydrothermal pretreatment
Biomass Project Research Center, Hiroshima Univ. 9
Temperature 130, 150, 170, 190, 200, 210, 240, 260 and 280 oC
Rubber wood powder 7 g
De-ionized water 63 g
0
50
100
150
200
250
300
0 50 100
Tem
per
atu
re [℃
]
Time [min]
130°C140°C150°C170°C190°C200°C210°C240°C260°C280°C
Temperature history for different target temperatures.
Experimental conditions
Biomass Project Research Center, Hiroshima Univ. 10
Rubber wood
residue 37% of market share export of
the world are from Thailand
Composition %
hemicelluloses 29
lignin 28
cellulose 39
ash 4
CHEMICAL CHARCTERISTICS
OF RUBBER WOOD RESIDUE
•United States Department of Agriculture, “Forage fiber analyses
(Apparatus, reagents, procedures, and some applications)”,
Agriculture Handbook, 379 (1970)
Feedstock
Biomass Project Research Center, Hiroshima Univ. 11
A B C
A B C D E
The samples after
hydrothermal pretreatment
at temperature on 130 (A),
140 (B), 150 (C), 170 (D),
and 190°C (E)
D E
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0 24 48
Yie
ld [
-]
Time of cellulase reaction [hour]
HC 130 °C
HC 140°C
HC 150°C
HC 170°C
HC 190°C
HC 200°C
HC 210°C
HC 240°C
HC 250°C
HC 260°C
HC 280°C
Time dependence of amount of glucose generated
from solid residue treated temperature 130–280oC,
10 wt% of concentration of raw material, treatment
time 0 min. (HC denote the solid sample from
hydrothermal pretreatment used with enzymatic
hydrolysis)
Products and saccharification
Biomass Project Research Center, Hiroshima Univ. 12
C G D
C*
1 k
3 k
2 k 4 k
) / exp( RT E A k ⊿ - =
C G D
C*
1
3
2 4
) / exp( RT E A k =
C Cellulose
C* Cellulose hydrolyzed by cellulase after pretreatment
G Glucose
D Decomposition products of glucose
C
C*
C
G
D
G
C*
C
DH
yd
roth
erm
al
pre
tre
atm
en
t
Cellu
las
e t
reatm
en
t
Reaction modeling
Biomass Project Research Center, Hiroshima Univ. 13
][][][
21 CkCkdt
Cd
][][*][][
423 GkCkCkdt
Gd
*][][*][
31 CkCkdt
Cd
][][
4 Gkdt
Dd
Preexponential
factor
[1/s]
Activation
energy
[kJ/mol]
k1 1.87×105 62.0
k2 2.02×107 87.7
k3 1.80×1018 222.2
k4 2.88×102 14.4
Reaction rate parameters
0
0.2
0.4
0.6
0.8
1
120 140 160 180 200 220 240 260 280 300
Temperature [oC]
Yie
ld [
-]
C C* G
D
Reaction rate parameters
Biomass Project Research Center, Hiroshima Univ. 14
0.00
0.20
0.40
0.60
0.80
1.00
140 150 160 170 180 190 200 210
Pretreatment Temp [℃]
Yie
ld [
-] C
C*
G
D
0.00
0.20
0.40
0.60
0.80
1.00
160 170 180 190 200 210 220 230 240
Pretreatment Temp [℃]
Yie
ld [
-] C
C*
G
D
Cabbage
EFB
0.00
0.20
0.40
0.60
0.80
1.00
150 160 170 180 190 200 210 220 230
Pretreatment Temp [℃]
Yie
ld [
-] C
C*
G
D
Kenaf
0
0.2
0.4
0.6
0.8
1
120140160180200220240260280300
Temperature [oC]
Yie
ld [
-]
C C* G
D
Comparison with other feedstocks
Biomass Project Research Center, Hiroshima Univ. 15
Model for the reactions in hydrothermal pretreatment
reactor was proposed.
The reaction parameter in the hydrothermal reactor for
rubber wood was successfully decided.
Reaction characteristics differs from feedstock to
feedstock.
緒言 Conclusions (reaction characteristics)
Biomass Project Research Center, Hiroshima Univ. 16
5 wt%YPD
10 mL rotary shaker, 30 ℃
incubator
Yeast
0.2 mL
Fermentatio
n inhibitor
Glucose and
ethanol
concentration
by HPLC
14 mL
vial
OD at 600 nm
緒言 Experiment for inhibitor effect clarification
Biomass Project Research Center, Hiroshima Univ. 17
Yeast
YPD medium (5.0 wt%)
Preculture
Formic acid
Acetic acid,
Furfural
5-HMF
Measuring time
Incubation temperature
S. cerevisiae *
10 mL
0.2 mL
0-45 mM
0-45 mM
0-45 mM
0-15 mM
36 h
30 oC
Inhibitor concentration
*Sigma-Aldrich (Type II)
緒言 Experimental conditions
Biomass Project Research Center, Hiroshima Univ. 18
μ=μmax KS
S
Xdt
dXμ
dt
dX
dt
dSk
(Monod equation)
XXoSo
k
SokXo
K
Xo
X
SokXo
K
1lnln)1(
μmax
1
X:Cell concentration
X0:Initial Cell concentration
S:Culture medium concentration
S0:Initial culture medium concentration
t: Incubation time
μmax: Maximum growth rate
K:Half medium concentration rate
k:
τ: Lag phase time
(1) Lag phase (2) Exponential growth phase
(3) Resting phase
)( τt
(1) (2) (3)
Time [h]
OD
at
600 n
m
τ
μmax
K
k
緒言 Cell growth model
Biomass Project Research Center, Hiroshima Univ. 19
Formic acid Acetic acid
0
0.1
0.2
0.3
0.4
0.5
0.6
0 10 20 30 40
OD
at
60
0 n
m [
-]
Time [h]
0 mM
15 mM
30 mM
45 mM 0
0.1
0.2
0.3
0.4
0.5
0.6
0 10 20 30 40O
D a
t 6
00
nm
[-]
Time [h]
0 mM 15 mM
30 mM
45 mM
緒言 Inhibitor effect on cell growth
Biomass Project Research Center, Hiroshima Univ. 20
Furfural 5-HMF
0
0.1
0.2
0.3
0.4
0.5
0.6
0 10 20 30 40
OD
at
60
0 n
m [
-]
Time [h]
0 mM
15 mM
30 mM
45 mM
0
0.1
0.2
0.3
0.4
0.5
0.6
0 10 20 30 40O
D a
t 6
00
nm
[-]
Time [h]
0 mM
5 mM
10 mM
15 mM
緒言 Inhibitor effect on cell growth
Biomass Project Research Center, Hiroshima Univ. 21
0.00
0.02
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
0 20 40
K/S
0 [
-]
Concentration [mM]
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
0 20 40
k/S0
[-]
Concentration [mM]
0.00
0.10
0.20
0.30
0.40
0.50
0.60
0 20 40
μm
ax [
1/h
]
Concentration [mM]
Formic acid Acetic acid Furfural 5-HMF
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
0 20 40
τ [h
]
Concentration [mM]
緒言 Monod parameter change by inhigibors
Biomass Project Research Center, Hiroshima Univ. 22
Formic acid Acetic acid
0 mM 15 mM 30 mM 45 mM
Ethanol
Glucose
0
0.05
0.1
0.15
0.2
0.25
0.3
0 10 20 30 40
Co
nce
ntr
atio
n [
mo
l/L]
Time [h]
0
0.05
0.1
0.15
0.2
0.25
0.3
0 10 20 30 40
Co
nce
ntr
atio
n [
mo
l/L]
Time [h]
緒言 Inhibitor effect on ethanol fermentation
Biomass Project Research Center, Hiroshima Univ. 23
Furfural 5-HMF
0
0.05
0.1
0.15
0.2
0.25
0.3
0 10 20 30 40
Co
nce
ntr
atio
n [
mo
l/L]
Time [h]
0
0.05
0.1
0.15
0.2
0.25
0.3
0 10 20 30 40
Co
nce
ntr
atio
n [
mo
l/L]
Time [h]
Furfural 0 mM 15 mM 30 mM 45 mM
5-HMF 0 mM 5 mM 10 mM 15 mM
Ethanol
Glucose
緒言 Inhibitor effect on ethanol fermentation
Biomass Project Research Center, Hiroshima Univ. 24
• The inhibitors used in this study slows cell growth
and final yeast concentration. Effect on
parameters were observed (μmax , k, τ).
• The inhibitors used in this study except acetic acid
decreases glucose consumption rate and ethanol
production rate for ethanol fermentation.
• Acetic acid affects cell growth but does not affect
ethanol production.
緒言 Conclusions (inhibitor effect)
Biomass Project Research Center, Hiroshima Univ. 25
This study was supported by the following funds.
Ministry of Education
Collaborating bodies
Tawatchai Charinpanitkul (Chulalongkorn University)
Staffs and Students Takuya Yoshida, Machi Kanna, Takashi Yanagida
Phacharakamol Petchpradab
Yuta Fukutomi
緒言 Acknowledgment
Biomass Project Research Center, Hiroshima Univ. 26
緒言
Thank you!!
See you at the European Biomass Conf. and Exhibition, 3-7 June 2013
Bella Center - Copenhagen, Denmark