Biofuels-Fundamental Research and Industrial Application

1. Shi-Zhong Li
Email:szli@tsinghua.edu.cn
shizhongli@ust.hk
Tel:+86 10 62772123
+852 34692229
Biofuels- Fundamental Research and
Industrial Application

2.  Volkswagen to promote
biofuel-powered cars
ahead of electric ones to
tackle carbon emissions,
new report suggests.
 COP21: Biofuels industry
calls for 15% biofuel in
global transport
Biofuels are one of the major technologies
available to decarbonise transport.

3.  All inner city bus lines run on
renewable fuels
 A total of 721 ethanol and 298
biogas buses in operation in
2013
 Reduced diesel use by
29 million litres / year
 Reduced fossil CO2 by
> 90 000 t / year
 Reduced PM 18.5 tons and NOx
185 tons
Net gain in Stockholm using ethanol
and biogas in buses
by courtesy of SL

4. In 2014, global
biodiesel
production was
26.64 million tons
to reduce 69.88
million tons of
automobile exhaust
particulates
7/28/2016

5. Hainan CONNC biodiesel plant with the
capacity of 60000 t/a
Biodesel standards (B100) was issued in May 2007.
Around 300 thousand tons of biodiesel were produced
annually in 2014, mostly used as solvent, mainly from waste
kitchen oil or residue from vegetable oil crushers.
B5 was in trial in Hainan Province in 2011, consumption tax
(5%) is removed.

6. However, the main problem to develop biodesel industry in China
is the shortage of raw materials, the feedstocks are focused on
woody oil plants, such as Jatropha curcas, Pistacia Chinensis,
Sapium sebiferum, and palm tree.
25Kg per spike of palm fruit
Palm trees are successfully
planted in Hainan Island

7. The Blunder Crop：
jatropha biofuels development
 Jatropha is realizing less than half its projected
yields in most projects, and less than a third of
optimistic estimates that led jatropha to be
labeled “the wonder crop”.
 Jatropha biodiesel uses 20,000 gallons of water
per gallon of fuel; soy 14,000
- University of Twente
http://www.utwente.nl

8.  FT diesel is Biomass
Integrated Gasification
Combined Cycle.
 CHOREN built a demon
plant with the capacity of
13000 tons of biodiesel
per year in April, since
there was no profit, Shell
quited the joint venture in
May 2012.
Second generation biofuel-FT diesel
CHOREN’s pilot facility of 200t/a

9. 性质 ethanol gasoline
Engine octane value 96 85
Research octane value 130 95
Octane value 113 90
In 2014, global bioethanol production was 75.23 million
tons to reduce 98.81 million tons of GHG and 17 thousand
tons of automobile exhaust particulates.

10. NASA confirms: biofuels do indeed burn cleaner
 FAME is not the answer, there is greater and greater risk for cross contamination
into the jet fuel stream- Pamela Serino, Defense Logistics Agency Energy
 However, waste cooking oil and jatropha can’t meet the demand of bio-ject fuel.
 Ethanol is the best feedstock to produce jet fuel via dehydration and
oligomerization, ethanol based jet fuel is listed in ASTMD7566-4

12. Only ethanol powered airplane in the world completes 10 years
 Compared with aviation gasoline (EMB-202), aviation ethanol (EMB-202A) spends 25% less
fuel on average. Furthermore, ethanol allows an increase of 7% in power, improving the
performance of the aircraft at takeoff, climb speed and maximum altitude
 By 2014, Embraer has sold 269 aircraft and 205 conversion kits for gasoline-powered planes
to fly with ethanol.

13. Japanese car giant Nissan Motor Co. is researching and developing what it calls a
solid oxide fuel-cell (SOFC)-powered system that runs on bio-ethanol electric power.
The new system – a world first for automotive use – features an e-Bio Fuel-Cell with
an SOFC power generator.
The e-Bio Fuel Cell generates electricity through the SOFC (power generator) using
bio-ethanol stored in the vehicle.
Nissan to develop world first ethanol-powered
electric car motor
Nissan to develop world first ethanol-powered electric car motor

14. A sign advertising E15, a gasoline with 15 percent of
ethanol, is seen at a gas station in Clive, Iowa, United
States, May 17, 2015.
Photo by Jim Young, Reuters
Final renewable fuel volumes
2014 2015 2016 2017
Cellulosic biofuel (million gallons) 33 123 230 n/a
Biomass-based diesel (billion
gallons)
1.63 1.73 1.90 2.00
Advanced biofuel (billion gallons) 2.67 2.88 3.61 n/a
Renewable fuel (billion gallons) 16.28 16.93 18.11 n/a
The US Environmental Protection Agency (EPA) has released the final
volume requirements for bioethanol under the Renewable Fuel
Standard (RFS) for the years 2014, 2015, and 2016. December 1, 2015

15. Logum is the world's first commercial ethanol pipeline, designed to become
Brazil's main ethanol transporter and bypass the country's lack of trains and poor
road infrastructure. The first phase of the project, the connection between the
town of Ribeirão Preto in São Paulo and the petrochemical plant in Paulinia, runs
for 206 kilometers and can carry up to 20 billion liters of ethanol per year.

16. BMW ramping up flex-fuel production in Brazil
BMW is now investing heavily in all corners of the world.
After focusing some of its attention on China, the
company is now expanding in the other extreme,
opening a new plant in Brazil. BY GABRIEL NICA 12 NOV.2014

17. 湖北省
河南省
安徽省
江苏省
山东省
黑龙江省
吉林省
辽宁省
河北省
四川省
重庆
广西省
江西省湖南省
河北省
山东省
湖北省
江苏省
黑龙江省
吉林省
辽宁省
安徽省
河南省
局部推广省份,
4省
全省封闭推广
省份, 6省
尚未推广省份,
21省（市、
区）
Part of the regions
Entire Regions
Fuel ethanol (E10) used area

18. Jilin Fuel Ethanol Co., Ltd.
Heilongjiang Huarun Ethanol Co., Ltd.
Henan Tianguan Group
Anhui Fengyuan BioChemicals Co., Ltd.
Guangxi Beihai Bioenergy Ltd.

19.  ethanol is shifting to non-food feedstock, such as sweet
sorghum, and Jerusalem artichoke (1.5 generation of bio-
ethanol)
 cellulosic ethanol (2nd generation of bio-ethanol) is
under R&D.
 China lowered the tariff on imports of ethanol to 5% from the
previous 30 % on 1 Jan 2010, and 0.6 million tons of ethanol
was imported from abroad in 2015.
China has limited arable land against a large population. To
ensure food security is a basic national policy. Fuel ethanol must
go for a non-food pathway though it was originated from grains.

20. Globally, biofuels
contribute about 3% of
transport energy, but
use significant
amounts of food
production to do so: in
recent years biofules
accounted for
11% of coarse grains
and vegetable oil use
 21% sugar cane use. Chris Woolston,Tough Characters: in search of hardy plants for biofuels
Looking for biofuel plants that can survive drought &
other harsh conditions
Abengoa's bioenergy arm files for bankruptcy in US
Feb. 25, 2016 http://biofuels-news.com/display_news/10234/abengoa039s_bioenergy_arm_files_for_bankruptcy_in_us/
Dilemma Status

21. 2014年9月3日DSM-POET纤维素乙醇厂开工典礼，
美国农业部长、能源部副部长、荷兰国王等参加。
 The $275 million factory ( 20 mgy cellulosic biofuel ) can accept 300,000
tons of biomass harvested from a 468 square-mile area.
 In a press conference in 2012, Broin said that the company cost of ethanol
production was below $3 per gallon but did not elaborate further. In
November 2009, the company projected a per-gallon cost of $2.35 at scale.
"With our first factory in the US making it possible,
we hope to bring it to China, too," Sijbesma told
China Daily on the sidelines of the World Economic
Forum in Tianjin on 12 Sept. 2014.

22.  The investments of 82 million liters of cellulosic ethanol per year reached US$ 265
million: US$ 190 million for the plant and US$75 million for a cogeneration unit.
 Currently, the plant is operating at a 20% load
Brazil’s first cellulosic ethanol plant Bioflex started operations
on 15 Sept. 2014 in the northern state of Alagoas.

23. RaÃzen to start 2G ethanol production in November 2014
The $92 million cellulosic biofuel factory (40 million litters per year)
"The challenge is to reach a trade price. There is still much to be learned," said
Pedro Mizutani, director of Operations of Raízen who estimates a further two or
three years for the 2G becomes equivalent to the first generation cost.
Cosan's net income plummeted in the third-quarter of 2014 to approximately 92.6%

24. KIOR declares bankrupcy on 9 Nov. 2014
 After several months of uncertainty, KIOR finally seeked Chapter 11 reorganization,
which basically means it is bankrupt on 9 Nov. 2014.
 A former cofounder said he tried to warn other board members about problems
with KiOR’s technology, as the company's Columbus-based refinery wasn't worked
as designed so KiOR idled the plant after stopping production in December.

25. DuPont's new cellulosic ethanol facility officially opened in Oct 2015,
but will ramp up production into 2016. This is the first plant of its kind
in the country - not the first cellulosic ethanol plant - but this one uses
proprietary processes the company developed

27. BA waste-to-jet fuel project fails to take off- 2016
 British Airways (BA) has announced that it has been forced to abandon plans to
turn landfill waste into green jet fuel, partly due to lack of government support.
 BA planned to turn 575,000 tonnes of household waste into gas. Enough green
fuel would have been produced to power all BA's yearly flights from London City
airport twice over.

28. 1st biofuels- ethanol from
corn and sugar cane
2nd biofules-
cellulosic ethanol 

Food crisis!
Cost-expensive!
Sweet sorghum can end the dilemma status of biofuels
1.5 generation-ethanol
from sweet sorghum
Cost-effective!


29.  Easy for big capacity
–More stable operation due to liquid processing
 The bagasse (residual of the stalks) can be sent to boilers
directly as fuel
 High energy consumption for process plant, due to juice
extraction consuming many power
 Around 5% sugar loss during juice extraction process
 More waste water produced due to:
–20% water added for juice abstraction
–Juice become waste water after fermentation and ethanol
abstracted
 Higher investment cost compare to solid fermentation for
similar capacity plant
Liquid Fermentation Using Juice
However, there is no current significant
bioethanol production based on sweet sorghum.
BNDES and CGEE, Sugarcane-based bioethanol : energy for
sustainable development. ISBN: 978-85-87545-27-5

30. What and Why is ASSF Technology ?
( Advanced Solid State Fermentation)
StalksStalks BagassesBagasses
 Short process
 Less fermentation time
 Less water consumption and less waste water
 Less energy consumption
 Less total investment cost
 Simple operation
FermentaterFermentater

31. Three basic requirement for ASSF: the excellent yeast,
automatically controlled fermenter, and sugar preservation
Du, et al. (2014) A Novel Wild-Type Saccharomyces cerevisiae
Strain TSH1 in Scaling-Up of Solid-State Fermentation of
Ethanol from Sweet Sorghum Stalks. PLoS ONE 9(4): e94480.
Wang, et al, Modeling of rotating drum bioreactor for
anearobic solid state fermentation. Applied Energy, 2010, 87:
2839-2845.

32. Chemistry and chemical engineering
 Unit operation
- solid state fermentation
- solid state distillation
 Mass transfer and heat transfer
- effect of particle size and water content on sugar transfer
- modeling of heat transfer and mass transfer during solid state fermentation
process
 Surface chemistry
- enzymes adsorption and the effect of lignin on enzymatic hydrolysis of
lignocellulose
 Molecular design
- the effect of hydrogen bond on enzymatic hydrolysis of lignocellulose

33.  Increase product yield
more active metabolic enzymes
higher production of metabolic enzymes, cellulases, b-glucosidase,
SLIC cloning to generate fusion protein to increase production of Hydrogen
in algae
 Heat tolerance
Increase fermentation temperature, more cost effective
Increase enzyme activity
 Increase resistance to inhibitory compounds
developing sulfite tolerant yeast strains by genetic cloning
 Mechanism of Action Studies
Molecular signaling pathway of recombinant microbes
Phylogenetic analysis
Compare differences in gene expression of consortiums
Genetic and metabolic engineering of microbes
to enhance biofuel production

34.  Isolation of bioet
hanol and bioga
s producing micr
obial consortia
 Next-generation
sequencing
 Bioinformatics
 Key strains
 Key genes
 Metabolic and
regulation
network
 Analysis the pathway of bioethanol and bio
gas production
 Construct the model of bioethanol and
biogas production
 Reconstruction of high bioethanol and
biogas-producing microbial consortium
Bioinformatics- the metagenomics and metatranscriptomics
study on microorganism for biofuel production

35. Research on ASSF in nm-dimension
 Unit Model of Mass Transfer of Fermentable Sugar
in sweet sorghum solid state fermentation
Metabolizing
rate of yeast
Solid-liquid
Extraction

36. Mass transfer kinetics of fermentable
sugar in xylem and pith
)e1()e1(* 21 -*
2
-*
1
tktk
CCC 
Transection of sweet
sorghum stalk without pill
Xylem
Pith
A modified kinetic model with structural coefficient
C1 and C2 are the structural coefficients for the sweet sorghum stalks.

37.  
 )*)02627.502681.098748.013728.0exp(exp(1
)*)80981.203678.001432.116645.0exp(exp(1
3212
3211
*
txxxC
txxxCC


)80981.203678.001432.116645.0exp( 3211  xxxk
)02627.502681.098748.013728.0exp( 3212  xxxk
x1:Particel size，mm; x2:osmatic pressure, ×103 kPa;
x3:temperature，℃。k1(min-1) and k2(min-1) are the mass
transfer rate of fermentable sugar in the pith and xylem,
respectively.
The mass transfer kinetic model of
fermentable sugar in sweet sorghum stalks

38. Kinetic Model Validation
Sugar extraction rate profiles, extraction conditions：
(a): x1：4 mm; x2 ：0 ×103KPa; x3：50 ℃; (b): x1：2 mm; x2：1.78×103KPa; x3：50 ℃
· experimental
—— predictable
· experimental
—— predictable
time time
Sugarextractionrate(%)
Sugarextractionrate(%)

39. 编号对应的酵母菌：
A：酿酒酵母TISTR5048
B：酿酒酵母Fleischmann
C：南阳酵母突变株NY-07017
D：酒精酵母 GJ2008 - 90 g/L
E：酒精酵母 GJ2008 - 230 g/L
F：酒精酵母 GJ2008 - 250 g/L
G：酒精酵母 GJ2008 - 270 g/L
H：酿酒酵母S-2002
I：酿酒酵母GGSF16
J：酿酒酵母苏-25
K：安琪酵母
L：N+离子束改造出发菌12#
M：N+离子束改造突变菌12#-70-11
N：N+离子束改造突变菌12#-90-A-1
O：酿酒酵母（Fermax yeast）
metabolizing process is the rate-controlling step of solid state
fermentation.
According to the unit model of solid state fermentation, the mass transfer
of fermentable sugar is significantly higher than the metabolizing rate of
sugar. So metabolizing process is the rate-controlling step of solid state
fermentation.

40. Research on ASSF in μm-dimension
because of the poor mass and heat transfer and
substrate specificity of sweet sorghum stem, we puts
forward requirements on yeast:
1.High temperature resistance
2.Product resistance (main products are ethanol
and ethyl ester )
3.Acid resistance
S.cerevisiae TSH1 screening of sweet sorghum is
proved that has excellent property mentioned above.
membrane
protein
transport
protein
In view of the excellent
property of TSH1,to gain
super yeast which has better
property via metabonomics
and evolutionary engineering
means will be of great
significance to sweet
sorghum ethanol industry, as
well as the future
development of the SSF
producing ethanol industry.
excellent characterization and molecular
modification of strains in SSF

41. Phylogenetic analysis using rRNA of a strain for solid
state fermentation
TSH1 is closely related to S. cerevisiae S288c. Phylogenetic tree reconstructed from the
neighbor-joining analysis of the 18S rDNA gene and 26S rDNA sequence of TSH1. The
bootstrap percentages over 50% (from 1000 bootstrap replicates) are shown. The
reference sequences were from the species type strains retrieved from GenBank under the
indicated accession numbers. The bars represent 0.01 substitutions per nucleotide position.
Du, et al. (2014) A Novel Wild-Type Saccharomyces cerevisiae Strain TSH1 in Scaling-Up of Solid-State
Fermentation of Ethanol from Sweet Sorghum Stalks. PLoS ONE 9(4): e94480.

42. Thermo-tolerant yeast strains
• Generated thermo-tolerant yeast strains S2-1 for solid state
fermentation by genomic shuffling between a good
fermentation strain T3 and a thermo tolerant strain F1.
Starting
glucose
(g/L)
Ending glucose
(g/L)
Ethanol
acumlated
(g/L)
Glucose
utilization
(%)
Ethanol yield
(%)
Theoretical
Ethanol Yield
(%)
30oC-24h
T3 132.39 0.00 56.19 100.00 83.22 83.22
F1 132.39 32.62 42.10 75.36 62.36 82.75
S2-1 132.39 0.00 56.69 100.00 83.96 83.96
40oC-24h
T3 132.39 0.43 55.45 99.68 82.12 82.39
F1 132.39 51.21 32.52 61.32 48.16 78.55
S2-1 132.39 4.00 54.91 96.98 81.32 83.86
43oC-40h
F1 132.39 71.53 22.56 45.97 33.42 72.70
S2-1 132.39 52.27 32.62 60.52 48.31 79.83
43oC-66h
F1 132.39 71.30 24.07 46.14 35.65 77.26
S2-1 132.39 40.99 38.54 69.03 57.08 82.68
45oC-72h S2-1 132.39 92.18 14.67 30.37 21.73 71.54
Feng, preliminary data

43. We have engineered 3 yeast strains which can produce ethanol at
43℃ with production yield of 79%.
差
异
表
达
基
因
数
量
73 12 74
T3-400 T3-600
83 17 62
T3-400 T3-600
TSH3
49 48 68
T1-400 T1-600
下调表达基因数量
52 12 50
T1-400 T1-600
上调表达基因数量
TSH1

44. Hardballmodel
ViscousBallmodel
Viscousrodmodel
Realbagaseparticle
Research on ASSF in mm-dimension
Mix at Radial direction

45. The mixture
of low
cohesive
particle with
different
long baffles
The mixture
of high
cohesive
particle with
different long
baffles
Enhanced mixture by baffles

46. Cohesive particle effects
a is the mixture of low cohesive particles
b is the mixture of high cohesive particles

47. The shapes of baffles affect the mixture
L- shape baffles with the same rotating direction bend has the
most positive effect to enhance the mixture.
L- shape baffle with the
same rotating direction bend
L- shape baffle with the
opposite rotating direction
bend
Straight baffle without any
bend

48. Condition Real Predict
MRT/s Particle
size（mm）
MRT/s Particle
size（mm）
5.4°4.26rpm 438.47 3 249.37 32
Calculation 410.86 3 248.41 32
Axial transfer simulation

49. 5 m3 (1.3k gallons)
10 L (2.6 gallons)
127 m3 (33.6k gallons)
555 m3 (132.1k gallons)
Flask Test
50 L (13.2 gallons)
2005
2006
2007
2007
2009
2011
49
250 L (66 gallons)
Zhang, et al, Scale-up of ethanol production from sweet sorghum using advanced solid-state fermentation. AIChE Annual
Meeting, Pittsburgh, PA, Nov 2, 2012
Wang, et al, Modeling of rotating drum bioreactor for anearobic solid state fermentation. Applied Energy, 2010, 87: 2839-2845.
Scale up of solid state fermentor

50. Dr. Buchanan, former
under secretary of USDA
visited the demon on 15
Sept. 2014.
A demon. plant under construction in Dongying,
Shandong Province, China

51. 50 % of fibrous residues (1.28t) for boiler fuel, 50% (1.28t)
to feed 1 cattle, and nutrition report is as the following
Raw material Dry
matter
Crude
Protein
content
Crude
fiber
Neutral
detergent
fiber
Acid
detergent
fiber
Crude
ash
Calcium Phosphate Total
energy
MJ/kg
Corn silage 24 1.47 4.59 9.89 5.76 1.34 0.06 0.06 16.44
Fermented
bagasse
(dry sample)
94.22 7.26 30.12 63.75 40.62 22.5 0.32 0.13 11.91

52. 52
1.5th Generation Biofuel – Sweet Sorghum Ethanol
Sweet sorghum
Pulverizer
Power system
Steam generator Continuous
distillation Rectifying
tower
Sorghum
rice
Livestock
Biogas
purifying
tower
Biogas tank
Advanced rotary
drum fermentor
Biogas reactor
A ‘close loop’: 1500ha/10000t ethanol and 4500t grain/6000
cattles/2.8 million Nm3 biogas and 60000t organic fertilizer
Sweet sorghum to
fuel & feed module

53. Grain/acre Stalks/acre USD remarks
Grain sorghum 150 bu (4.2t on) - 1050 $7/bu grain
Sweet sorghum 35.7 bu (1ton) 40 tons 1450 (250+1200) $30/ton stalk
Income of US farmers
Sweet sorghum can be planted all over the US, and help the US
to realize the goal of 35 billion gallons of ethanol by 2022.

54. 1 million ha are available to be used to produce 10 million tons of
ethanol competitively to supply the domestic need, and 2500 MW
electricity annually by 2025.

55. Primary agreement was
reached by Dr. Nazlee
Kamal and Dr. Shizhong
Li on 22 Aug. 2015:
 To establish the Sino-
Malaysia Join Research
Center for Biofuel & Bio-
based Products
 To start ethanol
production from sweet
sorghum by ASSF
 To start R&D on lactic
acid from sweet sorghum
stalks by ASSF.

56. On 29 July 2015, Dr. Yongyuth
Sawatdisawanee, director of Bureau of
Biofuel Development, Department of
Energy, Thailand, headed a delegation
to visit Tsinghua, and discussed the
potential collaboration:
√To introduce ASSF technology to
Thailand for cost-effective
production of ethanol
√To establish Sino-Thailand Joint
Research Center for Biofuel
 Now, Thailand’s ethanol is mainly from cassava and
molasses. Ethanol demand in 2018 is 2.96 billion liters.
 Khon Kaen University plans to develop sweet sorghum as
a new feedstock for ethanol.

58. Ethiopia is keen to establish bioethanol industry
A Ethiopian delegation visited demon
plant in Inner Mongolia, China
 If 1.6 million ha grain sorghum is replaced by
sweet sorghum, 10 million tons of ethanol can
be produced per year.
 2 million ha are available to be used to
produce 15 million tons of ethanol
competitively to supply the domestic need, 10
million tons of sorghum grain, and 20 billion
Kwh electricity annually.
 A new industry of more than $15 billion/a will
be built in 3-5 years in Ethiopia.
Dr Li met with Mr. Ato Girma,
former President of Ethiopia
Dr Li met with Mr. Hailemariam,
PM of Ethiopia.

59.  The South African government has introduced ASSF technology
from Tsinghua University to establish of sweet sorghum ethanol
industry of $12.25 billion/a in 5-10 years, including agriculture of
$2.25 billion and industry of $ 10 billion
 Dr. Li met with ministers of Department of Energy, Department of
Social development, and vice minister of Department of Finance in
Johnnesburg.

60. Heavy metal absorbing capacities of sweet sorghum
unit
Zn Cs As Cu Cd
Heavy metal content mg/kg 500.0 400.0 50.0 400.0 15.0
Ⅲ grade soil quality
standard
mg/kg 500 -- 30 400 1
Heavy metal
contents in
different parts of
sweet soghum
stalks
mg/kg
210.24 80.04 44.66 19.84 11.24
leaves 68.88 7.38 6.36 2.37 0.42
grains 60.76 23.13 12.91 5.73 3.24
Control stalks
mg/kg
18.62 0.21 0.64 3.22 0.16
leaves 8.71 0.06 0.14 0.49 0.02
grains 11.20 - - 1.92 0.088
Production yield:
75t fresh stalks, 1.2t leaves, 180kg grain per hectares

62. 桂阳
衡阳
株洲
赫山
The map of heavy metal contaminated
area in Xiang river valley
Sweet sorghum trial places
Sweet sorghum plantation
trial in Hunan

63. Cd contamination caused peasant unrest in 2006，in Xinma village, Zhuzhou city
Hunan province, the factory was shut down, however the abandoned field
（Cd>15mg/kg）is full of weeds. Sweet sorghum was planted in 2 Aug.
2013,harvested on 26 Dec. 2013, the highest stalk yield was more than 150t/ha.

64. 9 varieties with high Cd accumulating capacity
are selected from 229 sweet sorghum varieties
0.000
5.000
10.000
15.000
20.000
25.000
30.000
35.000
40.000
45.000
50.000
M94
M100
M8
M76
M28
M27
M51
M53
M91
M12
M61
M14
M67
M68
M64
M41
M57
M37
M38
M72
M98
M75
M42
Z120
Z119
Z92
Z84
Z124
Z55
Z107
Z15
Z20
Z27
Z52
Z90
Z102
Z35
Z82
Z54
Z74
Z83
Z71
Z75
Z26
Z100
Z63
Z80
Z87
Z95
植株
土壤
镉富集率
 19 of 100 varieties from USDA with high production yield of
more than 75t fresh stalk/ha, the highest is 150t/ha
 46 0f 129 varieties from Institute of Botany CAS with high
production yield of more than 75t fresh stalk/ha, the highest
is 136.5t/ha.

65. Cd recovery
From ash
20,000t ethanol
smashing/
ASSF
Bagasse for
burning
30 million Kwh to
grid（6MW）
3.6t Cd
320,000t sweet sorghum
stalks from 3000ha Cd
contaminated lands
 It’s reported that there are 1/6 arable lands are contaminated by heavy
metals in China now. There is no available technology to remediate these
lands cost-effectively.
Sweet sorghum can absorb heavy metals, and can be proceeded into
ethanol and electricity cost-effectively, and the heavy metals, such as Cd, will
be concentrated and recovered from ash simultaneously.
Sweet sorghum for fuel & power production is the unique practical and
cost-competitive technology to treat heavy metal contaminated croplands.

66. Composition of cellulosic ethanol cost:
1 feedstock 38-50%
2 preteatment 30-40%
TEA study of cellulosic ethanol conducted by NREL
Tao Ling et al, Process and technoeconomic analysis of leading pretreatment technologies for
lignocellulosic ethanol production using switchgrass. Bioresource Technology, 2011, 102.24: 11105

67. The combination of ASSF and
Alkaline pretreatment process for
ethanol production
34.8 Kg
Sweet Sorghum Stalk Crushing
Advanced
Solid State
Fermentation
Solid State
Distillation
1st Generation
Ethanol
Bagasse
Alkaline
Pretreatment
Enzymatic
Hydrolysis
Squeezing
c
Lignin
C5/C6 sugars
Co-fermentation
2nd Generation
Ethanol
One step
solid
liquid
1 ton Fresh Stalks
Containing 16% dry biomass
and 14% fermentable sugar
66 Kg
102 Kg
dry matters
One
One step
One
reactor
One step
Total Ethanol
100.8 Kg
Total Ethanol
100.8 KgMore than 7.5t ethanol/ha !
9.5 Kg
cellulose
hemicellulose
lignin

68. Energy input and output for novel cost-efficient integrated processes for
ethanol production from sweet sorghum stalks
68
Process Input (MJ) Output (MJ)
Smash Electricity 327.1
Preheat Electricity 80.6
Vapor 58.6
Seed culture Electricity 147.4
Vapor 58.6
Fermentation Electricity 209.2
Reaction
heat
502.0
Stripper Electricity 60.9
Steam 7,561.3 Ethanol 20,163.0
Certification Electricity 65.3 Lignin 5,386.5
enzymatic saccharification and
fermentation
Electricity 761.6
Distillation and separation Electricity 2,437.1 Ethanol 9,388.5
Other biorefinerya 380.8
Total 12,650.5 34,938.0
The energy input in other cellulosic ethanol processes is from 17,430 to 33,330 MJ/tonne
aZhu JY, Zhuang XS: Conceptual net energy output for biofuel production from lignocellulosic biomass through biorefining.Prog
Energ Combust 2012, 38:583–598.

69. 69
Conversion
ratio at 72h
Pretreatment Allomorph CrI Lattice
spacing
wettability Free
energy
Surface
area
1 [BMIM]Cl II 3
2 Ethylenediamine III 3 3 3 2
3 NaOH II 2 2 2 2 3
4 No treatment Iα 4 4 4 4 -
5 Glycerol Iβ 5 5 5 5 -
How does cellulose structure effect enzymatic hydrolysis
Ting Cui, et al. The Correlation between the Enzymatic Saccharification and the Multidimensional Structure of
Cellulose Changed by Different Pretreatments. Biotechnology for Biofuels, 2014, 7:134 doi:10.1186/s13068-
014-0134-6.

70. O % C% O/C %
Oextracted
%
Cextracted
%
O/Cextracted% Slig %
Ut 19.45 80.55 24.15 30.48 69.52 43.84 78.31
NaOH 32.52 67.48 48.19 35.80 64.20 55.76 54.47
Ca(OH)2 39.77 60.23 66.03 39.43 60.57 65.10 35.80
How does lignin effect enzymatic hydrolysis
Impact of AL and AIL content on the conversion of
cellulose(C) and hemicellulose (H) of sweet
sorghum bagasse (left is NaOH treatment , right is
Ca(OH)2 treatment )
NaOH
Ca(OH)2
Zhipei Yan, et al, Impact of Lignin Removal on the Enzymatic
Hydrolysis of Fermented Sweet Sorghum Bagasse. Applied
energy, 2015, http://dx.doi.org/10.1016/j.apenergy.2015.02.07

71. Integrated Biology for ONE STEP cellulosic
ethanol production
CBP by Mascoma, the US

72. The structure of bioethanol-
producing microbial-community
Members from Clostridium, Chelatococcus
and Tepidanaerobacter, are mostly enriched,
followed by Genus Paenibacillus and
Proteiniphilum.
Clostridium stercorarium degrades
polysaccharides and produces acetate,
ethanol, CO2, and H2, as well as minor
amounts of lactate and L-alanine
Clostridium thermocellum is a candidate
microorganism as it is capable of
hydrolyzing cellulose and fermenting the
hydrolysis products to ethanol
Acetivibrio cellulolyticus have a very
sophisticated cellulosome system that could
deconstruct cellulosic substrates
Paenibacillus sp. strain B39 is a novel
thermophilic, cellulosedegrading
bacterium
Ran Du, Jianbin Yan, Shizhong Li, et al, Cellulosic ethanol production
by natural bacterial consortia is enhanced by Pseudoxanthomonas
taiwanensis. Biotechnology for Biofuels, 2015, 8:10-19 DOI
10.1186/s13068-014-0186-7
Du, et al (2015), Optimization of cellulosic ethanol production
consortium via reconstruction of cellulolytic bacteria. (in preparation)

73. There are three primary challenges to cost-effective algae production:
 Algae de-watering stage is energy-intensive, and typically requires
chemical additives and expensive capital equipment.
 In order to extract oil , algae cell wall must be cracked. This is also
an energy-intensive process.
 It is critical that energy is recovered in every possible way, and
algae by-products must be harvested to achieve the best possible
energy balance.
Third generation biofuel-algae diesel

74. In 2009, European Algae Biomass
Association says commercialization 10-15
years away; US say 2-3 yrs: who’s right?
5-6,000 gallon per acre algae-to-energy production system

75. Algal Growth and Photo-H2 Production
from ASSF Wastewater
Green algae culture

76. A LED was Driven Directly by
a 40 mL Microalgae Culture
LED was lighted up by a single
hydrogen fuel cell which is
fueled by a single tube of
microalgae culture
Actually, in our electrochemical analysis, the voltage
generated by microalgal photo-H2 is as high as 1000 mV

77. Photo-H2 Driven Auto-flotation and its’ Application
in Harvesting of Oil Algae
Filamentous
culture Microscopy of
filaments
Auto-flotation of filaments was driven by photosynthetic H2
Application in harvests of
oil algae-Chlorella
1 hydrogen algae can float 6.69 oil algae

78. MOST-USDA Joint Research Center for Biofuels
The Jiont Center was established in Aug. 2008 under the Biofuels Cooperative
Activties between the Ministry of Science and Technology of P.R. China and the
Department of Agriculture of USA. In August, 2013 MOST and USDA signed an
agreement to extend the joint research to August, 2018.

79. Research Activities
 1.5 generation biofuel-ethanol from sweet sorghum stalks
using Advanced solid State Fermentation ASSF
– Scaling up of ethanol production using ASSF
– nm, µm, mm, and m scales of ASSF
 2nd generation biofuel-cellulose based biofuels
– Alkaline distillation pretreatment
– Cellulosic ethanol production using microbial consortia-
driven consolidated bioprocessing (CBP)
– An integrated low energy-consumption and cost-efficient
process for ethanol production from sweet sorghum
 4th generation biofuel- photosynthetic hydrogen production
using microalgae

80. Selected publications1. Jinhui Wang, Yinxin Li, Shizhong Li, et al, Lignin engineering through laccase modification: a promising field for energy plant improvement. Biotechnology for Biofuels, 2015, 8:145 DOI
10.1186/s13068-015-0331-y
2. Zhou, Z., Li, J, Li, S, et al. Enhancing mixing of cohesive particles by baffles in a rotary drum. Particuology (2015), http://dx.doi.org/10.1016/j.partic.2015.03.008
3. Ran Du, Jianbin Yan, Shizhong Li, et al, Cellulosic ethanol production by natural bacterial consortia is enhanced by Pseudoxanthomonas taiwanensis. Biotechnology for Biofuels, 2015, 8:10-
19 DOI 10.1186/s13068-014-0186-7
4. Juanjuan Feng, Shizhong Li, Yinxin Li, et al, High-throughput deep sequencing reveals that microRNAs play important roles in salt tolerance of euhalophyte Salicornia europaea. BMC Plant
Biotechnology, 2015, 15:63 DOI 10.1186/s12870-015-0451-3
5. Weitao Jia, Yinxin Li, Shizhong Li, et al, Restore Heavy Metal Contaminated Soil with Energy Plant. China Biotechnology, 2015, 35(1):88-95.
6. Yueying Mao, Jihong Li , Shizhong Li , Sandra Chang , Gang Zhao，The mass transfer of sugar in sweet sorghum stalks for solid-state fermentation process. Fuel, 2015, 144:90-95
7. Ting Cui, Jihong Li, Shizhong Li, et al. The Correlation between the Enzymatic Saccharification and the Multidimensional Structure of Cellulose Changed by Different Pretreatments.
Biotechnology for Biofuels, 2014, 7:134 doi:10.1186/s13068-014-0134-6.
8. Ming Chen, Jihong Li, Shizhong Li, et al, Auto-flotation of heterocyst enables the efficient production of renewable energy in cyanobacteria. Scientific Reports, 2014; 4:3998.
9. Ran Du, Jianbin Yan, Shizhong Li, et al, Optimization of ethanol production from NaOH pretreated solid state fermented sweet sorghum bagasse. Plos One, Published: April 15, 2014.DOI:
10.1371/journal.pone.0094480
10. Menghui Yu，Jihong Li, Shizhong Li, et al, A cost-effective integrated process to convert solid-state fermented sweet sorghum bagasse into cellulosic ethanol. Applied Energy, 2014,
115:331-336
11. Pei Pei, Chengming Zhang, ShizhongLi, et al, Optimization of NaOH pretreatment for enhancement of biogas production of banana pseudo-stem fiber using response surface methodology.
BioResources, 2014,9(3):5073-5087.
12. Jihong Li, Shizhong Li, et al. A novel cost-effective technology to convert sucrose and homocelluloses in sweet sorghum stalks into ethanol. Biotechnology for Biofuels 2013, 6:174-185
13. Shizhong Li, Guangming Li, Lei Zhang, et al. A demonstration study of ethanol production from sweet sorghum stems with advanced solid state fermentation technology. Applied Energy,
2013, 102: 260–265
14. Shaoxin Li, Jihong Li, Shizhong Li, et al. Study on enzymatic saccharification of Suaeda salsa as a new potential feedstock for bio-ethanol production. J. Taiwan Inst. Chem. Eng. 2013,44:
904–910
15. Fengcheng Li, Shuangfeng Ren, Shizhong Li, et al. Arabinose substitution degree in xylan positively affects lignocellulose enzymatic digestibility after various NaOH/H2SO4 pretreatments in
Miscanthus. Bioresource Technology 2013, 130 : 629–637
16. An Li, Yanan Chu, Shizhong Li*, et al. A pyrosequencing-based metagenomic study of methane-producing microbial community in solid-state biogas reactor. Biotechnology for Biofuels,
2013, 6 (3): 1-17
17. Chengming Zhang, Jihong Li, Shizhong Li*, et al. Alkaline pretreatment for enhancement of biogas production from banana stem and swine manure by anaerobic codigestion. Bioresource
Technology 2013, 149:353–358
18. Chen M, Zhiyu Zhang, Shizhong Li*, et al. Improving conversion efficiency of solar energy to electricity in cyanobacterial PEMFC by high levels of photo-H2 production, International
Journal of Hydrogen Energy 2013, 38: 13556-13563
19. Tian Meng; Liu Xiaoling; Li Shizhong*，Characteristics of solid-state anaerobic co-digestion for converting banana stalk and manure to biogas，Transactions of Chinese Society of
Agriculture Engineering，2013，29（7）：177-184
20. Z. Lewis Liu, Scott A. Weber, Shi-Zhong Li, et al. A new β-glucosidase producing yeast for lower-cost cellulosic ethanol production from xylose-extracted corncob residues by simultaneous
saccharification and fermentation. Bioresource Technology 2012, 104: 410–416
21. Jihong Li, Shizhong Li, Chenyu Fan, et al, The mechanism of poly(ethylene glycol) 4000 effect on enzymatic hydrolysis of lignocellulose. Colloids and Surfaces B: Biointerfaces 2012, 89:
203–210
22. Han Bing, Fan Guifang, Li Shizhong, et al. Comparison of three sugar feedstocks and two yeast strains in ethanol production by solid state fermentation, Transactions of Chinese Society of
Agriculture Engineering，2012,28(5):201-206
23. Han Bing, Li Jihong, Li Shizhong, et al., RESEARCH ON SWEET SORGHUM STALKS STORAGE TECHNOLOGIES FOR FUEL ETHANOL PRODUCTION, Acta Energiae Solaries Sinica，2012,33 (10):
1719-1723
24. Quanzhou Feng, Shizhong Li, et al, Evaluation on glucose-xylose co-fermentation by a recombinant Zymomonas mobilis strain. Chinese Journal of Biotechnology, 2012 28(1):37-47.
25. Zhao Yunfei, Liu Xiaoling, Li Shizhong*, et al, Effects of organic substance mixing ratios on methane bioconversion through high-solids anaerobic co-fermentation，China Environmental
Science，2012,32(6)：1110-1117
26. Fu Xiaofen, Li Shizhong, Wei Ming, The production of L(+)-lactic acid through the fermentation of glucose and xylose by thermophilic lactobacillus, Industrial Microbiology, 2012，6:52-57
27. Lei Zhang, Jihong Li, Shizhong Li, et al, Challenges of Cellulosic Ethanol Production from Xylose – Extracted from Corncob Residues. BioResources, 2011, 6(4):4302-4316
28. Ran Du, Shizhong Li, et al, Using a microorganism consortium for consolidated bioprocessing cellulosic ethanol production. Biofules, 2011, 2 (5):569-575.
29. Xiaoling Liu, Shizhong Li*, et al, Study on high-solids mesophilic-anaerobic digestion of waste activated sludge to produce biogas. ACTA SCIENTIAE CIRCUMSTANTIAE, 2011, 31(5): 955-963
30. Erqiang Wang, Shizhong Li, Ling Tao, Modeling of rotating drum bioreactor for anearobic solid state fermentation. Applied Energy, 2010, 87: 2839-2845.
31. Geng X, Li SZ, et al, Studies on the process parameters of solid state fermentation for fuel ethanol production from sweet sorghum stalks and pilot test. Acta Energiae Solaries Sinica 2010;
31(2):257–262

81. Thank
You
“It always
seems
impossible
until it’s
done”.