Shi-Zhong Li

Email:szli@tsinghua.edu.cn

shizhongli@ust.hk

Tel:+86 10 62772123

+852 34692229

Biofuels- Fundamental Research and Industrial Application

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.

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 by29 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 ethanoland biogas in buses

by courtesy of SL

In 2014, global biodiesel production was 26.64 million tons to reduce 69.88 million tons of automobile exhaust particulates

7/28/2016

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.

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

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

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

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.

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

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.

Japanese car giant Nissan Motor Co. is researching and developing what it calls asolid 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 withan SOFC power generator.The e-Bio Fuel Cell generates electricity through the SOFC (power generator) usingbio-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

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 finalvolume requirements for bioethanol under the Renewable FuelStandard (RFS) for the years 2014, 2015, and 2016. December 1, 2015

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.

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

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

Jilin Fuel Ethanol Co., Ltd.

Heilongjiang Huarun Ethanol Co., Ltd.

Henan Tianguan GroupAnhui Fengyuan BioChemicals Co., Ltd.

Guangxi Beihai Bioenergy Ltd.

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.

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 USFeb. 25, 2016 http://biofuels-news.com/display_news/10234/abengoa039s_bioenergy_arm_files_for_bankruptcy_in_us/

Dilemma Status

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.

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.

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," saidPedro Mizutani, director of Operations of Raízen who estimates a further two orthree 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%

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.

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

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.

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 sorghumCost-effective!

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

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

Three basic requirement for ASSF: the excellent yeast, automatically controlled fermenter, and sugar preservation

Du, et al. (2014) A Novel Wild-Type Saccharomyces cerevisiaeStrain 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.

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

Increase product yieldmore active metabolic enzymeshigher production of metabolic enzymes, cellulases, b-glucosidase, SLIC cloning to generate fusion protein to increase production of Hydrogen in algae

Heat toleranceIncrease fermentation temperature, more cost effective

Increase enzyme activity

Increase resistance to inhibitory compoundsdeveloping sulfite tolerant yeast strains by genetic cloning

Mechanism of Action StudiesMolecular signaling pathway of recombinant microbesPhylogenetic analysisCompare differences in gene expression of consortiums

Genetic and metabolic engineering of microbes to enhance biofuel production

Isolation of bioethanol and biogas producing microbial 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 andbiogas production

Reconstruction of high bioethanol and

biogas-producing microbial consortium

Bioinformatics- the metagenomics and metatranscriptomics

study on microorganism for biofuel production

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

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.

)*)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

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

Sugar

ext

ract

ion r

ate

(%

)

Sugar

ext

ract

ion r

ate

(%

)

编号对应的酵母菌：A：酿酒酵母TISTR5048B：酿酒酵母FleischmannC：南阳酵母突变株NY-07017D：酒精酵母 GJ2008 - 90 g/LE：酒精酵母 GJ2008 - 230 g/LF：酒精酵母 GJ2008 - 250 g/LG：酒精酵母 GJ2008 - 270 g/LH：酿酒酵母S-2002I：酿酒酵母GGSF16J：酿酒酵母苏-25K：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#-70-11N：N+离子束改造突变菌12#-90-A-1O：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.

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 resistance2.Product resistance (main products are ethanol and ethyl ester )3.Acid resistanceS.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

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.

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(%)

TheoreticalEthanol 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

We have engineered 3 yeast strains which can produce ethanol at43℃ 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

Ha

rd b

all m

od

el

Vis

co

us

Ba

ll mo

de

l

Vis

co

us

rod

mo

de

l

Re

al b

ag

as

e p

artic

le

Research on ASSF in mm-dimension

Mix at Radial direction

The mixture of low cohesive particle with different long baffles

The mixture of high cohesive particle with different long baffles

Enhanced mixture by baffles

Cohesive particle effects

a is the mixture of low cohesive particlesb is the mixture of high cohesive particles

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

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

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

20072007

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

Dr. Buchanan, former under secretary of USDA visited the demon on 15 Sept. 2014.

A demon. plant under construction in Dongying, Shandong Province, China

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 Drymatter

Crude Proteincontent

Crudefiber

Neutral detergent fiber

Aciddetergentfiber

Crude ash

Calcium Phosphate Total energyMJ/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

1.5th Generation Biofuel – Sweet Sorghum Ethanol

Sweet sorghum

Pulverizer

Power systemSteam generator Continuous

distillation Rectifying tower

Sorghumrice

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

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.

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.

Primary agreement was reached by Dr. NazleeKamal and Dr. ShizhongLi 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.

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.

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.

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.

Heavy metal absorbing capacities of sweet sorghum

unitZn 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/kg18.62 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

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 plantationtrial in Hunan

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.

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

M100M8

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.

Cd recoveryFrom ash

20,000t ethanol

smashing/ ASSF

Bagasse for burning

30 million Kwh togrid（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.

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

The combination of ASSF and Alkaline pretreatment process for

ethanol production

34.8 Kg

Sweet Sorghum Stalk CrushingAdvanced Solid State

Fermentation

Solid StateDistillation

1st Generation Ethanol

Bagasse

AlkalinePretreatment

Enzymatic Hydrolysis

Squeezing

c

Lignin

C5/C6 sugarsCo-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

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 327.1Preheat Electricity 80.6

Vapor 58.6Seed culture Electricity 147.4

Vapor 58.6Fermentation Electricity 209.2

Reaction heat

502.0

Stripper Electricity 60.9Steam 7,561.3 Ethanol 20,163.0

Certification Electricity 65.3 Lignin 5,386.5enzymatic saccharification and

fermentationElectricity 761.6

Distillation and separation Electricity 2,437.1 Ethanol 9,388.5Other 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/tonneaZhu JY, Zhuang XS: Conceptual net energy output for biofuel production from lignocellulosic biomass through biorefining.Prog Energ Combust 2012, 38:583–598.

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.

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

Integrated Biology for ONE STEP cellulosic ethanol production

CBP by Mascoma, the US

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, cellulosedegradingbacterium

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/s13068-014-0186-7Du, et al (2015), Optimization of cellulosic ethanol production consortium via reconstruction of cellulolytic bacteria. (in preparation)

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

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

Algal Growth and Photo-H2 Production from ASSF Wastewater

Green algae culture

A LED was Driven Directly bya 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

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

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.

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

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-y2. 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.2015.03.0083. 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/s13068-014-0186-74. 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/s12870-015-0451-35. 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, 144:90-957. 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.009448010. 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, 6:174-18513. 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: 260–26514. 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:

904–91015. 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 : 629–63716. 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): 1-1717. 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, 149:353–35818. 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: 13556-1356319. 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）：177-18420. 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: 410–41621. 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:

203–21022. 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):201-20623. 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):

1719-172324. 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)：1110-111726. Fu Xiaofen, Li Shizhong, Wei Ming, The production of L(+)-lactic acid through the fermentation of glucose and xylose by thermophilic lactobacillus, Industrial Microbiology, 2012，6:52-5727. Lei Zhang, Jihong Li, Shizhong Li, et al, Challenges of Cellulosic Ethanol Production from Xylose – Extracted from Corncob Residues. BioResources, 2011, 6(4):4302-431628. 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): 955-96330. 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

Thank

You

“It always seems

impossible until it’s done”.