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 Metabolic Regulation and
Metabolic Engineering for Biofuel
and Biochemical Production
 Metabolic Regulation and
Metabolic Engineering for Biofuel
and Biochemical Production

Kazuyuki Shimizu
Institute for Advanced Biosciences
Keio University, Yamagata
Japan

p,
p,
A SCIENCE PUBLISHERS BOOK
A SCIENCE PUBLISHERS BOOK
CRC Press
Taylor & Francis Group
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 Preface

The increasing economic growth and the prosperity has accelerated

worldwide with the increasing demand on the energy, mostly generated
from fossil fuels since the beginning of the industrial revolution in the mid
eighteenth century. This has brought about the rapidly increasing global
warming, caused by the emission of the green-house gas such as CO2,
resulting in disastrous climate change, where this problem will become
crucial even to the level of human survival by the end of this century.
It is, therefore, the time to take action to protect further global warming
by employing the alternative clean renewable sources of energy. Among
the renewable energy sources such as nuclear, solar, wind energies, etc.,
bioenergy seems to be the most promising alternative source of renewable
energy in the long range future.
This book intends to explain the current status and future perspectives
for biofuel and biochemical production from biomass, and motivates the
innovation for future energy problem. Moreover, attention is focused on
the production of biofuels and biochemicals directly from atmospheric CO2
by photosynthetic microorganisms. In particular, it is important to create
new approaches that bring innovation or revolution for bio-based energy
generation. The center for this is the metabolism of organisms. Although
many papers have been published on the production of biofuels and
biochemicals by metabolic engineering or synthetic biological approaches,
less attention is focused on the metabolic regulation. Understanding the
metabolism or metabolic regulation mechanisms in response to pathway
modification or genetic modulation is critical for the innovative design of
the cell metabolism for efficient biofuel production providing a solution to
the future energy problems. Thus the present book intends to explain the
metabolic regulation mechanisms including catabolic regulation prior to
the metabolic engineering practices. The metabolic engineering practices
are subdivided into two categories: one, using heterotrophic bacteria, yeast,
and fungi, and second, the photoautotrophic autotrophic microorganisms
such as algae and cyanobacteria. Finally, brief explanation is given for the
systems biology approach and modeling.
 Contents

Preface v
Notations xiii

1. Background 1
1. Current Status of Global Warming and Action Plan 2
2. Attempts to Reduce Energy Consumption 5
3. Alternative to Petroleum-based Fuels 6
3.1 Nuclear power energy 6
3.2 Wind energy 7
3.3 Solar energy 8
3.4 Other sources of renewable energy 8
3.5 Grid energy storage and future perspectives of 9
renewable energy sources
4. Bio-based Energy Generation for the Reduced CO2 Emission 9
5. Biofuel and Biochemical Production from Biomass 11
6. Brief Summary and the Outlook of the Book 12
References 14
2. Pretreatment of Biomass 16
1. Introduction 17
2. Various Pretreatments 20
2.1 Physical pretreatment 22
2.2 Biological pretreatment 22
2.3 Chemical pretreatment 24
2.4 Physiochemical pretreatment 26
3. Simultaneous Saccharification and Fermentation (SSF) 28
4. Consolidated Biomass Processing (CBP) 30
5. Concluding Remarks 32
References 32
3. Transport of Nutrients and Carbon Catabolite Repression for 38
the Selective Carbon Sources
1. Introduction 39
2. Variety of Regulation Mechanisms 40
viii Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

3. Porin Proteins in the Outer Membrane and their Regulation 42

4. Transport of Carbohydrates and PTS 46
5. Carbohydrate Uptake by Various PTSs and without PTS 50
6. Nitrogen PTS 52
7. Carbon Catabolite Repression for the Selective Carbon 53
Source Uptake
8. CCR in other Bacteria than E. coli 58
9. Concluding Remarks 61
References 61
4. Catabolite Regulation of the Main Metabolism 70
1. Introduction 70
2. Regulation of the Glycolytic Flux 72
3. Enzyme Level Regulation of the Glycolysis 74
4. Regulation of Pyruvate Kinase 79
4.1 Regulation of Pyk expression by isozymes 79
4.2 Effects of Pyk mutation on the metabolism 79
5. Transcriptional Regulation of the Glycolysis 84
6. Overflow Metabolism and the Oxidative Stress Regulation 86
7. Constraint on ATP Production by Respiration 90
8. Respiratory Pathways and the Competition with 91
Catabolic Transport
9. Coordination of the Metabolism by cAMP-Crp at Higher 94
Catabolic Rate
10. Carbon Catabolite Repression 96
10.1 Carbon catabolite repression in E. coli 97
10.2 Catabolite regulation in Corynebacteria 99
10.3 Catabolite regulation in Baccili 100
10.4 Catabolite regulation in Clostridia 100
11. Heteroginity of the Cell Population and CCR 101
12. Carbon Storage Regulation 103
12.1 Carbon storage regulation in E. coli 104
12.2 Carbon storage regulation in a variety of organisms 106
13. Concluding Remarks 109
References 110
5. Metabolic Regulation in Response to Growth Environment 126
1. Introduction 127
2. Nitrogen Regulation 127
3. Sulfur Regulation 131
4. Phosphate Regulation 132
5. Metal Ion Regulation and Oxidative Stress Regulation 133
6. Redox State Regulation 135
7. Acid Shock Response 137
 Contents ix

8. Heat Shock Stress Response 139

9. Cold Shock Response 140
10. Solvent Stress Regulation 141
11. Osmoregulation 142
12. Biofilm, Motility by Flagella, and Quorum Sensing 143
13. Concluding Remarks 145
References 147
6. Metabolic Engineering for the production of a Variety of 155
Biofuels and Biochemicals
1. Introduction 156
2. Typical Fermentation 159
2.1 Pyruvic acid fermentation 161
2.2 Lactic acid fermentation 162
2.3 Acetic acid fermentation 186
3. Production of TCA Cycle Intermediates 187
3.1 Citric acid production 189
3.2 Malic acid production 192
3.3 Fumaric acid fermentation 200
3.4 Succinic acid production 206
4. Diol Fermentation 207
4.1 Propanediol production 207
4.1.1 1,3-propanediol production 208
4.1.2 1,2-propanediol production 208
4.2 Butandiol production 212
4.2.1 2,3-butandiol production 212
4.2.2 1,4-Butandiol production 215
4.2.3 1,3-Butandiol production 215
5. Other Organic Acid Fermentation 216
5.1 Itaconic acid fermentation 217
5.2 Glucaric acid production 219
5.3 Muconic acid production 220
5.4 Adipic acid production 222
6. Amino Acids and Related Fermentation 222
6.1 Shikimic acid fermentation 223
6.2 Phenylalanin and biofuels production by 225
modulating Csr
6.3 Production of L-Tyrosine and its derived compounds 225
7. Isoprenoid, Polyketide, and Alkanoid Production 225
8. Biofuel Production 226
8.1 Hydrogen production 227
8.2 Ethanol fermentation 228
8.3 Higher alcohol production 239
8.4 Fatty acid fermentation 241
x Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

9. Tolerance to Solvent Stresses 242

10. Concluding Remarks 244
References 245
7. Biofuel and Biochemical Production by Photosynthetic 276
Organisms
1. Introduction 277
2. Candidate Photosynthetic Micro-organisms for Biochemical 278
and Biofuel Production
3. Metabolism of Photosynthetic Microorganisms 280
3.1 Brief overview of the metabolism of photosynthetic 280
organisms
3.2 Metabolism of algae 281
3.3 Metabolism of cyanobacteria 283
3.4 Nitrogen regulation and carbon storage regulation in 288
cyanobacteria
4. Metabolic Engineering of Photosynthetic Microorganisms 290
4.1 Metabolic engineering of algae 290
4.2 Metabolic engineering of cyanobacteria 291
4.2.1 Hydrogen production 292
4.2.2 Ethylene production 293
4.2.3 Ethanol production 296
4.2.4 Isopropanol production 297
4.2.5 Lactic acid production 297
4.2.6 Acetone production 300
4.2.7 Butanol production 301
4.2.8 Fatty acid and fatty alcohol production 302
4.2.9 Isoprene production 306
4.2.10 Diols production 308
(a) 1,2-propanediol 308
(b) 1,3-propanediol 309
(c) 2,3-butanediol 310
4.2.11 PHB production 311
4.2.12 3-hydroxybutyrate production 314
4.2.13 Other metabolite production 315
4.3 Efficient CO2 fixation 317
5. Systems Biology Approach and Modeling of the Metabolism 318
6. Cultivation Methods 320
7. Harvesting of Algal Biomass 320
8. Downstream Processing 321
9. Concluding Remarks 323
References 324
 Contents xi

8. Systems Biology Approach and Modeling for the 341

Design of Microbial Cell Factories
1. Introduction 342
2. Flux Balance Analysis and its Extensions 343
3. Kinetic Modeling and Incorporation of Metabolic Regulation 346
4. Modeling of the main Metabolism for Catabolite Regulation 348
5. Importance of the Modeling for the main Metabolic Pathways 350
6. Metabolic Regulation Mechanisms to be Incorporated in the 352
Kinetic Model
7. Modeling for the Metabolism under Oxygen Limitation 361
8. Concluding Remarks 363
References 364
Appendix A 373
Index 375
About the Author 387
 Notations

(General)
ABC transporter ATP-binding cassette transporter
CBB cycle Calvin-Benson-Bassham cycle
CBP consolidated bioprocess
CCR carbon catabolite repression
ETC electron transport chain
FFA free fatty acid
PP pathway pentose phosphate pathway
TCA cycle tri-carboxylic acid
PS photo system
PTS phosphotransferase system
ROS reactive oxygen species
SHF separate hydrolysis and fermentation
SSF simultaneous saccharification and
fermentation
(Metabolite)
AA amino acid
AcAcCoA acetoacetyl coenzyme A
ACC 1-aminocyclopropane 1-carboxylic
acid
AcCoA acetyl coenzyme A
AcP acetyl phosphate
ADP adenosine diphosphate
ADPG adenosine 5’-diphospho glucose
ALA amino-levulinic acid
Alg alginine
xiv Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

AMP adenosine monophosphate

Asp aspartic acid
ATP adenosine triphosphate
BDO butanediol
1,3BPG 1,3-bisphosphoglycerate
CHA chorismic acid
CIT citrate
DAHP 3-deoxy-D-arabinoheptulosonate
7-phosphate
DHA dihydroxyacetone
DHAP dihydroxyacetone phosphate
DHQ 3-dehydroqunic acid
DHS 5-dehydro shikimic acid
DMAPP demethylalyl diphosphate
E4P erythrose 4-phosphate
EPSP 5-enolpyruvate-shikimate-3-
phosphate
F6P fructose 6-phosphate
FBP fructose 1,6-bisphosphate
Fru fructose
FUM fumarate
G1P glucose 1-phosphate
G6P glucose 6-phosphate
GABA ɤ-amino butyric acid
GAP glyceraldehyde 3-phosphate
GL3P glycerol 3-phosphate
Glc glucose
Glu glutamic acid
Gln glutamine
GOX glyoxylate
GSH glutathinon
GTP guanosine triphosphate
HA hydroxyalkanoate
 Notations xv

HB hydroxybutyric acid
IA itaconic acid
ICI isocitrate
KDPG 2-keto-3-deoxy-6-phosphogluconate
αKG α-ketoglutaric acid
KMBA 2-keto-4-methyl-thiobutyric acid
Lys lysine
MA muconic acid
MAL malate
2MB 2-methyl-1-butanol
MEP 2-C-methyl-D-erythritol 4-phosphate
MG methylglyoxal
MQ menaquinone
MVA mevalonic acid
OAA oxaloacetate
OSP oseltamivir phosphate
PDO propanediol
2PG 2-phosphoglycerate
3PG 3-phosphoglycerate
PEP phosphoenol pyruvate
PHA polyhydroxy alkanoate
PHB polyhydroxybutyrate
PLA polylactic acid
PMA plymalic acid
6PG 6-phosphogluconate
PGA poly-β-1,6-N-acetyl-D-glucosamine
PPY phenylpyruvate
PQ plastoquinone
PQQ pyrroloquinone
PUFA polyunsaturated fatty acid
PYR pyruvate
R5P ribose 5-phosphate
R5PI ribose phosphate isomerase
xvi Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

Ru5P ribulose 5-phosphate

RuBP ribulose 1,5-bisphosphate
SA shikimic acid
S7P sedoheptulose 7-phosphate
Ser serine
SUC succinate
TAG triacylglycerol
Trp tryptophane
Tyr tyrosine
UQ ubiquinone
UQOH ubiquinol
X5P xylulose 5-phosphate
XOS xylo-oligosaccharide

(Protein)
ACC acyl-acyl carrier protein
Ack acetate kinase
ACP acyl carrier protein
Acs acetyl coenzyme A synthetase
ADC (Adc) acetoacetate decarboxylase
ADH alcohol dehydrogenase
Adk adenylate kinase
ADSL adenylosuccinate lyase
AHL acyl-homoserine lactone
AI arabinose isomerase
Ald aldolase
ALDC 2-acetolactate decarboxylase
ALDH aldehyde dehydrogenase
ALS acetolactate synthase
AOR aldehyde oxidoreductase
ASL alginino-succinate lyase
AspP adenosine diphosphate sugar
pyrophosphatase
 Notations xvii

AtoB acetyl transferase

BDH butanediol dehydrogenase
CA carboxylic anhydrase
cAMP cyclic adenosine monophosphate
CcpA catabolite control protein A
CimA citramalate synthase
Cra catabolite repressor/ acticvator
Crp cAMP receptor protein
CS citrate synthase
Csr carbon storage regulator
Ctf CoA transferase
Cya adenylate cyclase
DXS 1-deoxy-D-xylulose 5-phosphate
synthase
EI enzyme I
EII enzyme II
EFE ethylene forming enzyme
Eno enolase
F1PK D-fructose-1-phosphate kinase
FAA fumarylacetoacetase
FAS fatty acid synthase
FHL formate hydrogenlyase
Fnr fumarate nitrate reductase
Fum fumarase
G6PDH glucose 6-phosphate dehydrogenase
GADC glutamate decarboxylase
GAPDH glyceraldehydes 3-phosphate
dehydrogenase
GDH glutamate dehydrogenase
GlgA glycogen synthase
GlgB glycogen branching enzyme
GlgC ADPG phosphorylase
GlgP glycogen phosphorylase
xviii Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

Glk glucokinase
Glx glyoxalase
GlgX glycogen debranching enzyme
GlpK glycerol kinase
GlyDH glycerol dehydrogenase
GOGAT Glutamate synthase (glutamine
oxoglutarate aminotransferase)
GS glutamine synthetase
Hbd 3-hydoxybutyryl-CoA
dehydrogenase
HPr histidine-phosphorylatable protein
HPrK HPr kinase
Hxk hexokinase
Hyd hydrogenase
ICDH isocitrate dehydrogenase
Icl isocitrate lyase
IspS isoprene synthase
KDC ketoacid decarboxylase
Kivd 2-keto-acid-decarboxylase
LADH L-arabitol 4-dehydrogenase
LALDH lactaldehyde dehydrogenase
LDH lactate dehydrogenase
Lrp leusine responsive regulatory protein
LXR L-xylulose reductase
MarR multiple antibiotic resistant regulator
MDH malate dehydrogenase
Mez(Mae) malic enzyme
Mgs methylglyoxal synthase
MIOX myo-inositol oxygenase
MS malate synthase
Nac nitrogen assimilation control protein
NR nitrogen regulator
Pck phosphoenolpyruvate caroxykinase
 Notations xix

PDC pyruvate decarboxylase

PDH pyruvate dehydrogenase
PDOX propanediol oxidoreductase
Pfk phosphofructokinase
PGDH 6-phosphogluconate dehydrogenase
Pgi phosphoglucose isomerase
Pgm phosphoglucomutase
Pgk phosphoglycerate kinase
PhaA β-ketothiolase
PhsB acetoacetyl-CoA reductase
PhaC polyhydroxy alkanoate (PHA)
synthase
PI3K phosphoinositide 3-kinase
Ppc phosphoenolpyruvate carboxylase
Pps phosphoenolpyruvate synthase
PRK phosphoribulokinase
Pta phosphotransacethylase
Pyc pyruvate carboxylase
Pyk pyruvate kinase
RhaD rhamnose dehydrogenase
RK ribulokinase
Rpe ribulose 5-phosphate 4-epimerase
Rpi ribulose 5-phosphate isomerase
RubisCo ribulose 1,5-bisphosphate
carboxylase
SADH secondary alcohol dehydrogenase
SDH succinate dehydrogenase
SOD superoxide dismutase
Tal transaldolase
TE thioesterase
Ter trans-2-enoyl-CoA reductase
Thl thiolase
Tkt transketolase
xx Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

Udh urinate dehydrogenase

XDH xylitol dehydrogenase
XI xylulose isomerase
XK xylulokinase
XR xylose reductase
 1
Background

ABSTRACT
The increasing economic growth and the prosperity has been accelerated
worldwide with the increasing demand on the energy mostly generated
from fossil fuels since the beginning of the industrial revolution in the mid
eighteenth century. This has brought about rapid global warming caused
by the emission of the greenhouse gases such as CO2, resulting in the
disastrous climate change, and this problem will become endangering even
to the level of human survival by the end of this century. It is, therefore,
the time to take action to prevent further global warming by employing
the alternative clean renewable sources of energy. Among the renewable
energy sources such as nuclear, solar, wind energies, etc., bioenergy seems
to be the most promising alternative source of renewable energy in the
long range future.
The so-called 1st generation biofuels have been produced from corn starch
and sugarcane in USA and Brazil. However, this causes a problem of the
so-called “food and energy issues” as the production scale increases. The
2nd generation biofuels production from lignocellulosic biomass has thus
been paid more attention recently. However, it requires energy intensive
pre-treatment for the degradation of lignocellulosic biomass. The 3rd
generation biofuels production from photosynthetic organism such as
cyanobacteria and algae has also come under attention, but the cell growth
rate and thus the productivity of the fuels is significantly low. The typical
processes for biofuel and biochemical production from biomass include
pre-treatment of biomass, saccharification, fermentation, and separation
of the dilute fermentation broth followed by purification.
Keywords
Global warming, renewable energy, pretreatment, lignocellulosic biomass,
biofuel, biochemical, 1st generation biofuel, 2nd generation biofuel, 3rd
generation biofuel, low carbon society
2 Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

1. Current Status of Global Warming and Action Plan

The increasing economic growth and the prosperity have been accelerated
worldwide with the increasing demand on the energy generated from
mostly fossil fuels since the beginning of the industrial revolution in the mid-
eighteenth century. This has been promoted by the steam powered trains
and ships, and then the internal combustion engines significantly changed
the human’s life style and production system in relation to transportation
and industrial production. Nowadays, many people live comfortable
lives spending much electricity for air conditioning and freezing of food,
etc. Many people use cars and airplanes to move across the countries and
the continents spending more and more fossil fuels as sources of energy.
Unlike the beginning of the industrial revolution, where the population
of the world was 700 million, the current population is 7 billion and
is estimated to grow to 9 billion by 2,050, and even about 10 billion by
2,100 (Lee 2011). Unlike the developed countries, the population growth
is more eminent in the developing countries like Asia and Africa, where
the economic growth is accelerating year by year, caused by promotion
of industrialization to catch up to the standard of posh and comfortable
living of the developed countries. The International Energy Agency (IEA)
has projected that the world’s energy demand will increase from about
12 billion ton oil equivalents (t.o.e.) in 2,009 to either 18 billion t.o.e. or 17
billion t.o.e. by 2,035 under the current policies or new policies scenarios,
respectively (International Energy Agency 2011). In association with this,
carbon dioxide (CO2) emissions are expected to increase from 29 giga tons
(Gt) per year to 43 Gt/year or 36 Gt/year under the current policy and new
policies, respectively. This creates significant climate-change risks in either
policies, and we are now facing a risk to even human survival by the end of
this century. Namely, we need another industrial revolution for the energy
sources to be affordable, accessible, and sustainable. Energy efficiency and
conservation, as well as decarbonizing the energy sources are essential to
the revolution (Chu and Majumdar 2012).
International Panel on Climate Change (IPCC) keeps warning the
global society on global warming caused by greenhouse gases (GHGs) such
as CO2 based on the accumulating data and the reliable prediction model.
IPCC asks world societies to make decisions to take actions and to invest
in the reduction of CO2 emissions caused mostly by human activities. This
may be also considered from the point of view of future cost caused by the
severe climate change due to global warming. Namely, the global warming
may cause serious local climate change as well as the rise in the sea level,
which will severely damage the societies worldwide. In fact, every year
we are experiencing disastrous climate change, and it seems to grow more
and more severe. We have recently experienced disastrous phenomena as
 Background 3

extreme heat waves, droughts due to segregated rain falls, wild fires in
the broad forest, melting glaciers, etc. These phenomena may have been
caused largely by the global warming most likely due to the accumulation
of GHGs in the atmosphere, mainly CO2 from the burning of fossil fuels. In
2013, a big news was broadcasted throughout the world warning that the
CO2 level in the earth’s atmosphere had passed over 400 ppm (parts per
million) for the first time in several million years. As shown in Fig. 1, the
CO2 level periodically changed in accordance with the repetition of warming
and cooling of the earth probably due to the effect of black body irradiation
from the surface of the sun since 400,000 years ago. However, the sharp
increase in the CO2 level caused by the human activities after industrial
revolution becomes eminent, and this is becoming more and more severe.
The international society has recognized the importance of this
problem since the late 20th century. The international political activities
against climate change began at the Rio Earth summit in 1992, where Rio
convention included the adoption of the United Nations Framework
Convention on Climate Change (UNFCCC). This convention attempted
to take action for stabilizing atmospheric concentrations of GHGs to avoid
“dangerous anthropogenic interference with climate system.” The UNFCCC
has currently a near universal membership of 195 parties including both
developed and developing countries.

Figure 1. Global change in CO2 concentration and the temperature on earth (https://
stevengoddard.wordpress.com/2014/09/26/climate-is-not-difficult/, http://cdiac.ornl.gov/
trends/co2/recent_mauna_loa_co2.html).
4 Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

The annual conference of parties (COP) began in Berlin in 1995,

followed by several conferences such as COP3 in Kyoto, where the Kyoto
Protocol was adopted, COP11 in Montreal, where the action plan was
determined, COP17 in Durban, where the Green Climate Fund was created.
Unfortunately, some large CO2 producing countries denied signing the
agreement of Kyoto Protocol, and the efficient action against climate change
has been unsuccessful. As mentioned above, however, the global warming
steadily proceeds, where the glaciers and the iceberg in the north-pole
are melting, causing sea level to rise, and causing drastic climatic change
worldwide, and the atmosphere is becoming more and more humid
due to vaporization of sea water at an unprecedented rate caused by the
temperature rise. The international societies strongly recognize the current
and future critical situations caused by CO2 emission from fossil fuels.
In 2015, COP21 (also known as the 2015 Paris Climate Conference)
aimed to achieve a legally binding and universal agreement on climate,
where the conference was attracted about 50,000 participants including
25,000 official delegates from government, intergovernmental organizations,
united nation (UN) agencies, non-governmental organizations (NGOs) and
civil societies. Then a historic agreement was signed to combat climate
change and unleash actions and investment towards a low carbon, resilient
and sustainable future. Each county must set individual target and must
attain it, where the action result will be checked every 5 years, but this is
rather effort target without legal constraint, even though this is a big one
step towards preventing further global warming. The aim of the universal
agreement is to keep the global temperature rise well below 2 degrees
Celsius (2ºC) during this century, and take strong action to limit the
temperature rise even further to 1.5ºC above pre-industrial level.
Now the developed countries face the problem of promoting
industrialization without or much less emission of CO2 with efficient
transportation and production systems, or may have to change the life
styles drastically, while the developing countries may have to keep
industrialization without or less amount of emission of CO2 with the aid
of technology transfer and investment from the developed countries.
Namely, green based innovation and revolution is necessary to attain such
target and to realize a low carbon society. Now is the time to pay attention
to innovations that provide an entirely new energy system including
transportation and stationary systems together with energy generation
systems. It is also of importance to investigate the ways of energy efficiency
and the integration of energy sources with electricity transmission,
distribution and storage (Chu and Majumdar 2012).
Currently, petroleum-oriented liquid fuels are the main source of energy
in the transportation infrastructure throughout the world. The geographical
distribution of petroleum resources changes as the new resources such as
 Background 5

shale oil field are found and accessed by new technologies for discovery
and production. The distribution of petroleum production does not coincide
with the place of the demand. This means that fair amount of fuels must
be transported spending energy for this purpose. For example, about 2.690
billion tons of oil were consumed, where 1.895 billion tons of crude oil and
0.791 billion tons of refined products crossed national borders (BP Statistical
review of the world 2012). This also brings along with itself political problem
of national-security for energy, where many countries are forced to import
oils from the limited oil producing countries.
The overall loss caused by the increase in the number of heat waves,
floods, wildfires, droughts, and storms may be estimated to be over $150
billion per year (Munich Re 2012), where these phenomena may possibly be
caused by the climate change (Cambridge University Press 2012). The cost
of renewable energy is rapidly becoming competitive with other sources
of energy.

2. Attempts to Reduce Energy Consumption

Improvements in energy efficiency can contribute for the reduction of fuel
usage. One idea is to use light-weight materials for vehicles. Some attempts
have been made for ultra-high tensile strength steels, carbon-fiber reinforced
composite materials, aluminium and magnesium alloys, and polymers
(Gibbs et al. 2012). The potential of reducing the weight of vehicles such as
cars and airplanes has already been shown without sacrificing safety, and
it is expected to reduce more about 20–40% (Powers 2000). It is estimated
that for every 10% weight reduction of the vehicle, 6–8% of improvement
is expected in fuel consumption (Holmberg et al. 2012). Moreover, the
reduction of the friction loss in vehicles may also contribute for the efficient
energy usage, where several attempts have been made for the development
of cost effective technologies such as tyres, braking and waste-heat energy
recovery, etc. (Holmberg et al. 2012), where Rankin cycle may be utilized
to convert waste heat to work by low-cost and high efficiency solid state
thermoelectric systems (Yang and Caillat 2006).
Plug-in hybrid electric vehicles (PHEVs) or electric vehicles (EVs)
have the opportunity to displace a significant amount of liquid fuel use
in transportation, and reduce GHG emission. Such vehicles are becoming
competitive, a mass-market car with the internal combustion engine (ICE)
using liquid transportation fuel. The high efficiency of fuel-cell-powered
electric vehicles may become a potential option in the near future, where
there are inherent volumetric energy density issues for hydrogen-gas
storage. The supply infrastructure and a low-carbon source of hydrogen
are challenged in several countries. The technology advances in shale-gas
production may give impact on the transportation system. In addition to
6 Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

the direct use of natural gas as a fuel, low-cost natural gas may give local
reforming or hydrogen filling stations in the near future, where delivery
to local filling stations may be made through high-pressure gaseous tube
trailers in practice. Due to the low cost of natural gas as compared to
petrol, natural-gas vehicles may become more widely used, where liquefied
natural gas (LNG)-powered trucks and trains may be considered for the
long distance transportation (Chu and Majumdar 2012).

3. Alternative to Petroleum-based Fuels

The time-scale and the mix of the current energy sources is shown in Fig. 2,
where the ratios have been significantly changed after industrial revolution
in mid 1800s.

Figure 2. Change in energy usage after industrial revolution. World energy consumption by
source, based on Vaclav Smil estimates from Energy Transitions: History, Requirements and
Prospects together with BP Statistical Data for 1965 and subsequent. https://ourfiniteworld.
com/2012/03/12/world-energy-consumption-since-1820-in-charts/.

3.1 Nuclear power energy

Nuclear power may play an important role in decarbonizing the energy

production, where the percentage of nuclear energy usage is about 14% of
the total electric power generated in 2009 (International Energy Agency
 Background 7

2010), although this percentage dropped to about 12% in 2011, mainly

caused by the less usage in Japan due to Fukushima disaster and Germany
(BP 2012, Ruhl 2011). After the Fukushima nuclear disaster in 2011, all the
nuclear power plants were eventually shut down in Japan. This disaster
affected the energy policies of various countries, where some countries keep
running the nuclear power plants with careful caution; some have opted
to reverse their decision on nuclear energy, or accelerating planned-phase-
outs (Chu and Majumdar 2011). In some developing countries, however,
the electricity shortage is critical, which prevents the comfortable life and
industrialization, and thus the nuclear energy is among the top priority
for such countries with the risk of safety. The most significant problem in
using nuclear energy is treating the spent fuel before disposal, which is still
contaminated with highly radioactive particles. Moreover, the numbers of
old nuclear power plants (say more than about 40 years old) are increasing,
and the cost of amortizing plants is not low. Hence the humans are yet to
learn to control the nuclear energy.

3.2 Wind energy

Significant improvement has been made in turbines, blades and gearboxes,

and the height of the wind towers to reduce the cost. As the height and
size of the wind turbines increase, more and more mechanical stresses are
imposed on the gearbox, blades and tower. Direct-drive wind turbines can
convert the slow rotary motion directly into alternating current electrical
power using electromagnetic generators, which consist of permanent
magnets composed of rare-earth metals. The dependence of the materials
on the rare-earth gives a problem of unforeseen future availability, since
more than about 90% of the rare-earth metals are located in China (US
Department of Energy: DOE 2012).
Offshore wind farms are attractive in the sense that they can be placed
in near-proximity to cities, and yet far enough away to mitigate local
opposition. However, it is not easy to construct a marine structure that
can withstand the harsh marine environment for many years, and thus
the cost remains high, which prevents its application in practice (Chu and
Majumdar 2012).
As the scales and the number of wind turbines increase, the effect of
such towers with fans on the aerodynamics, and the effect of high frequency
sound on the human health must be carefully analyzed. Such problems
could be avoided by installing the wind towers far from the population
area, but another problem of energy loss during the long-distance transfer
of energy may become eminent.
8 Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production

3.3 Solar energy

Although the installation cost for solar panels are becoming lower due to
mass production, the apparent limitations are the locations and the areas
to be used, and the dependence on the sunny weather. The costs may be
further decreased by increasing the efficiency of the solar cells. The ultimate
efficiency limit in the conversion of sunlight energy to electric energy is
determined by a Carnot heat engine, where the limit is estimated to be
about 94% based on the temperature of the sun’s black body at 5,800 degree
Kelvin (K) and the cell temperature of about 350 K. The Shockley-Queisser
limit is a well-known efficiency limit of single-junction solar cells, which
suffer from spectrum losses, recombination and black-body radiation, where
the limit of the silicon is about 37%, but may actually be low at around
29% in practice (Swanson 2005). This efficiency may be more improved by
considering the above theoretical upper bound.
The solar thin films are made of crystalline substances, and
polycrystalline-silicon photovoltaics, direct-gap semi-conductors such
as cadmium telluride and copper-indium gallium-selenide (CIGS).
Cadmium-telluride thin films are used, but there is yet significant room
for improvement. Improvement of solar-module efficiency may also come
from multi-function cells that capture a large fraction of the solar spectrum,
multi-exciton generation, multi-photon absorption or photon up-conversion
and light concentration (Chu and Majumdar 2012).
Solar flat panels may be also replaced by the concentrated solar
photovoltaics, where the main cost of the system is the mirrors, lenses, and
their tracking system rather than solar cell at the focal point of the system
(Angel and Olbert 2011).
In order to convert direct current to alternating current electricity and
to integrate solar power into the grid, power electronics is crucial (US
Department of Energy 2012).

3.4 Other sources of renewable energy

Hydrolytic power energy has long been employed in particular at

mountainous regions, where water fall is necessary to catch the energy from
the river. Although this may be important for the protection of flooding
by the dam capacity during strong heavy rain, etc., this is limited to the
mountainous locations.
Geothermal energy is another source of energy associated with
volcanos, but the installation of the power plant may dry up the hot springs,
and may disturb the geothermal environment.
Another random document with
no related content on Scribd:
 Launcelot.—Nay, indeed, if you had your eyes, you might fail of
the knowing me: it is a wise father that knows his own child. Well, old
man, I will tell you news of your son: give me your blessing. Truth will
come to light; murder cannot be hid long,—a man’s son may; but, in
the end, truth will out.
Gobbo.—Pray you, sir, stand up: I am sure you are not Launcelot,
my boy.
Launcelot.—Pray you, let’s have no more fooling about it, but give
me your blessing: I am Launcelot, your boy that was, your son that
is, your child that shall be.
Gobbo.—I cannot think you are my son.
Launcelot.—I know not what I shall think of that: but I am
Launcelot, the Jew’s man; and I am sure Margery your wife is my
mother.
Gobbo.—Her name is Margery, indeed: I’ll be sworn, if thou be
Launcelot, thou art mine own flesh and blood. Lord, worship’d might
he be! What a beard hast thou got! thou hast got more hair on thy
chin than Dobbin, my fill-horse, has on his tail.
Launcelot.—It should seem, then, that Dobbin’s tail grows
backward: I am sure he had more hair of his tail than I have on my
face, when I last saw him.
Gobbo.—Lord, how art thou chang’d! How dost thou and thy
master agree? I have brought him a present. How ’gree you now?
Launcelot.—Well, well; but, for mine own part, as I have set up my
rest to run away, so I will not rest till I have run some ground. My
master’s a very Jew: give him a present! give him a halter: I am
famish’d in his service; you may tell every finger I have with my ribs.
Father, I am glad you are come: give me your present to one Master
Bassanio, who, indeed, gives rare new liveries: if I serve not him, I
will run as far as God has any ground.—O rare fortune! here comes
the man:—to him, father, for I am a Jew, if I serve the Jew any
longer.
—Act II, Scene II, Lines 29-104.
 HAMLET’S DECLARATION OF FRIENDSHIP

Hamlet. What ho! Horatio!

Horatio. Here, sweet lord, at your service.

Hamlet. Horatio, thou art e’en as just a man

As e’er my conversation coped withal.

Horatio. O, my dear lord,—

Hamlet. Nay, do not think I flatter;

For what advancement may I hope from thee
That no revenue hast, but thy good spirits,
To feed and clothe thee? Why should the poor be flatter’d?
No, let the candied tongue lick absurd pomp,
And crook the pregnant hinges of the knee
Where thrift may follow fawning. Dost thou hear?
Since my dear soul was mistress of her choice
And could of men distinguish, her election
Hath sealed thee for herself; for thou hast been
As one, in suffering all, that suffers nothing,
A man that fortune’s buffets and rewards
Hast ta’en with equal thanks: and blest are those
Whose blood and judgment are so well commingled
That they are not a pipe for Fortune’s finger
To sound what stop she pleases. Give me that man
That is not passion’s slave, and I will wear him
In my heart’s core, ay, in my heart of hearts,
As I do thee.

—From Act III, Scene 2.

OTHELLO’S APOLOGY
[The speech calls for great dignity, ease, and power, in both speech
and manner.]
Most potent, grave, and reverend signiors,
My very noble and approved good masters,
That I have ta’en away this old man’s daughter,
It is most true; true, I have married her:
The very head and front of my offending
Hath this extent, no more. Rude am I in my speech,
And little bless’d with the soft phrase of peace;
For since these arms of mine had seven years’ pith,
Till now some nine moons wasted, they have used
Their dearest action in the tented field,
And little of this great world can I speak,
More than pertains to feats of broil and battle,
And therefore little shall I grace my cause
In speaking for myself. Yet, by your gracious patience,
I will a round unvarnish’d tale deliver
Of my whole course of love; what drugs, what charms,
What conjuration, and what mighty magic,—
For such proceeding I am charg’d withal,—
I won his daughter.
...
Her father loved me; oft invited me;
Still question’d me the story of my life,
From year to year,—the battles, sieges, fortunes,
That I have pass’d.
I ran it through, even from my boyish days,
To the very moment that he bade me tell it:
Wherein I spake of most disastrous chances,
Of moving accidents by flood and field,
Of hair-breadth scapes i’ the imminent deadly breach,
Of being taken by the insolent foe
And sold to slavery, of my redemption thence
And portance in my travels’ history:
...

This to hear
Would Desdemona seriously incline:
But still the house-affairs would draw her thence;
Which ever as she could with haste despatch,
She’d come again, and with a greedy ear
Devour up my discourse: which I observing,
Took once a pliant hour, and found good means
To draw from her a prayer of earnest heart
That I would all my pilgrimage dilate,
Whereof by parcels she had something heard,
But not intentively: I did consent,
And often did beguile her of her tears,
When I did speak of some distressful stroke
That my youth suffer’d. My story being done,
She gave me for my pains a world of sighs:
She swore, in faith, ’twas strange, ’twas passing strange,
’Twas pitiful, ’twas wondrous pitiful:
She wish’d she had not heard it, yet she wish’d
That heaven had made her such a man: she thank’d me,
And bade me, if I had a friend that loved her,
I should but teach him how to tell my story,
And that would woo her. Upon this hint I spake:
She loved me for the dangers I had pass’d;
And I lov’d her that she did pity them.
This only is the witchcraft I have used.

THE SEVEN AGES

[This is a succession of purely imaginative ideas which the voice
should touch lightly. In this speech one meets always the question of
impersonation: shall the mewling infant, the whining schoolboy, the
sighing lover and the rest be imitated by the reader? It is in better
taste not to impersonate these seven characters beyond certain
almost imperceptible hints which the gayety of Jaques’s mind might
naturally throw off.]

All the world’s a stage,

And all the men and women merely players:
They have their exits and their entrances;
And one man in his time plays many parts,
His acts being seven ages. At first the infant,
Mewling and puking in the nurse’s arms:
And then the whining schoolboy, with his satchel
And shining morning face, creeping like snail
Unwillingly to school. And then the lover,
Sighing like furnace, with a woeful ballad
Made to his mistress’ eyebrow. Then a soldier,
Full of strange oaths and bearded like the pard,
Jealous in honor, sudden and quick in quarrel,
Seeking the bubble reputation
Even in the cannon’s mouth. And then the justice,
In fair round belly with good capon lined,
With eyes severe and beard of formal cut,
Full of wise saws and modern instances;
And so he plays his part. The sixth age shifts
Into the lean and slipper’d pantaloon,
With spectacles on nose and pouch on side;
His youthful hose, well saved, a world too wide
For his shrunk shank; and his big manly voice,
Turning again toward childish treble, pipes
And whistles in his sound. Last scene of all,
That ends this strange eventful history,
Is second childishness and mere oblivion,
Sans teeth, sans eyes, sans taste, sans everything.

—“As You Like it,” Act II, Scene 7.

SOLITUDE PREFERRED TO COURT LIFE

Duke S. Now, my co-mates and brothers in exile,

Hath not old custom made this life more sweet
Than that of painted pomp? Are not these woods
More free from peril than the envious court?
Here feel we but the penalty of Adam.
The season’s difference, as the icy fang
And churlish chiding of the winter’s wind,
Which, when it bite and blows upon my body,
Even till I shrink with cold, I smile and say
’Tis no flattery; these are counselors
That feelingly persuade me what I am.
Sweet are the uses of adversity,
Which, like the toad, ugly and venomous,
Wears yet a precious jewel in his head;
And this our life, exempt from public haunt,
Finds tongues in trees, books in the running brooks,
Sermons in stones, and good in everything.
I would not change it.

Amiens. Happy is your grace,

That can translate the stubbornness of fortune
Into so quiet and so sweet a style.

...

Duke S. Come, shall we go and kill us venison?

And yet it irks me the poor dappled fools,
Being native burghers of this desert city,
Should in their own confines with forked heads
Have their round haunches gor’d.

—“As You Like It,” Act II.

THE POTION SCENE

Scene: Juliet’s Chamber

(Enter Juliet and Nurse, who bears wedding garments.)

Juliet (looking at garments).

Ay, those attires are best; but, gentle nurse,

I pray thee, leave me to myself to-night;
For I have need of many orisons
To move the heavens to smile upon my state,
Which, well thou knowest, is cross and full of sin.
 (Enter Lady Capulet.)

Lady Capulet.

What are you busy, ho? need you my help?

Juliet.

No, madam; we have cull’d such necessaries

As are behoveful for our state to-morrow:
So please you, let me now be left alone,
And let the nurse this night sit up with you;
For, I am sure, you have your hands full all,
In this so sudden business.

Lady Capulet (crossing and kissing Juliet on the forehead).

Good night;
Get thee to bed and rest, for thou hast need.

(Exit Lady Capulet with nurse.)

Juliet (looking after them).

Farewell! God knows when we shall meet again.

I have a faint cold fear thrills through my veins,
That almost freezes up the heat of life:
I’ll call them back again to comfort me. (Runs to R.)
Nurse! What should she do there?
My dismal scene I needs must act alone.
Come, vial. (Takes vial from bosom.)
What if this mixture do not work at all?
Shall I be married then to-morrow morning?
No, no! (draws dagger) this shall forbid it.

(Lays dagger on table.)

Lie you there. (To vial.)

What if it be a poison, which the friar
Subtly hath ministered to have me dead,
Lest in this marriage he should be dishonored
Because he married me before to Romeo?
I fear it is; and yet, methinks, it should not,
For he hath still been tried a holy man.

(Puts vial in bosom.)

How if, when I am laid into the tomb,

I wake before the time that Romeo
Come to redeem me? there’s a fearful point!
Shall I not then be stifled in the vault,
To whose foul mouth no healthsome air breathes in,
And there die strangled ere my Romeo comes?
Or, if I live, is it not very like,
The horrible conceit of death and night,
Together with the terror of the place,—
As in a vault, an ancient receptacle,
Where, for these many hundred years, the bones
Of all my buried ancestors are packed;
Where bloody Tybalt, yet but green in earth,
Lies festering in his shroud; where as they say,
At some hours in the night spirits resort; ...
O, if I wake, shall I not be distraught,
Environed with all these hideous fears?
And madly play with my forefathers’ joints?
And pluck the mangled Tybalt from his shroud?
And, in this rage, with some great kinsman’s bone,
As with a club, dash out my desperate brains?
O, look! methinks I see my cousin’s ghost
Seeking out Romeo, ...
Stay, Tybalt, stay!—
Romeo, I come! (Drawing out vial—then cork.)
This do I drink to thee.

(Throws away vial. She is overcome and sinks to the floor.)

—From “Romeo and Juliet,” Act IV, Scene 3.

 BANISHMENT SCENE
SCENE III, A ROOM IN THE PALACE
(Enter Celia and Rosalind.)
Cel. Why, cousin; why Rosalind;—Cupid have mercy;—Not a
word?
Ros. Not one to throw to a dog.
Cel. No, thy words are too precious to be cast away upon curs,
throw some of them at me; come, lame me with reasons.
Ros. Then there were two cousins laid up; when the one should be
lamed with reasons, and the other mad without any.
Cel. But is all this for your father?
Ros. No, some of it for my father’s child: O, how full of briars is this
working-day world!
Cel. They are but burrs, cousin, thrown upon thee in holiday
foolery; if we walk not in the trodden paths, our very coats will catch
them.
Ros. I could shake them off my coat; these burrs are in my heart.
Cel. Hem them away.
Ros. I would try; if I could cry hem, and have him.
Cel. Come, come, wrestle with thy affections.
Ros. O, they take the part of a better wrestler than myself.
Cel. Is it possible, on such a sudden, you should fall into so strong
a liking with old Sir Rowland’s youngest son?
Ros. The duke my father lov’d his father dearly.
Cel. Doth it therefore ensue, that you should love his son dearly?
By this kind of chase, I should hate him, for my father hated his
father dearly; yet I hate not Orlando.
Ros. No ’faith, hate him not, for my sake.
Cel. Why should I not? Doth he not deserve well?
 Ros. Let me love him for that; and do you love him, because I do:
Look, here comes the duke.
Cel. With his eyes full of anger.
(Enter Duke Frederick, with Lords.)
Duke F. Mistress, despatch you with your safest haste, and get
you from our Court.
Ros. Me, uncle?
Duke F. You, cousin, within these ten days if thou be’st found so
near our public court as twenty miles, thou diest for it.
Ros. I do beseech your grace, let me the knowledge of my fault
bear with me: if with myself I hold intelligence, or have acquaintance
with mine own desires; if that I do not dream, or be not frantic (as I
do trust I am not), then, dear uncle, never so much as in a thought
unborn, did I offend your highness.
Duke F. Thus do all traitors, if their purgation did consist in words,
they are as innocent as grace itself: let it suffice thee, that I trust thee
not.
Ros. Yet your mistrust cannot make me a traitor: tell me, whereon
the likelihood depends.
Duke F. Thou art thy father’s daughter, there’s enough.
Ros. So was I, when your highness took his dukedom; so was I,
when your highness banish’d him: treason is not inherited, my lord:
or, if we did derive it from our friends, what’s that to me? my father
was no traitor: then, good my liege, mistake me not so much, to think
my poverty is treacherous.
Cel. Dear sovereign, hear me speak.
Duke F. Aye, Celia; we stay’d here for your sake. Else had she
with her father rang’d along.
Cel. I did not then entreat to have her stay, it was your pleasure,
and your own remorse; I was too young that time to value her, but
now I know her; if she be a traitor, so am I: we still have slept
together; rose at an instant, learn’d, play’d, eat together;

And wheresoe’er we went, like Juno’s swans,

Still we went coupled, and inseparable.

Duke F. She is too subtle for thee; and her smoothness,

Her very silence, and her patience,
Speak to the people and they pity her.
Thou art a fool: she robs thee of thy name;
And thou wilt show more bright, and seem more virtuous,
When she is gone: then open not thy lips;
Firm and irrevocable is my doom
Which I have pass’d upon her; she is banish’d.

Cel. Pronounce that sentence then on me, my liege;

I cannot live out of her company.

Duke F. You are a fool:—You, niece, provide yourself;

If you outstay the time, upon my honor,
And in the greatness of my word, you die.

(Exeunt Duke Frederick and Lords.)

Cel. O my poor Rosalind: whither wilt thou go?

Wilt thou change fathers? I will give thee mine.
I charge thee, be not thou more griev’d than I am.

Ros. I have more cause.

Cel. Thou hast not, cousin,

Pr’ythee, be cheerful: know’st thou not, the duke
Hath banish’d me his daughter?

Ros. That he hath not.

Cel. No? hath not? Rosalind lacks then the love

Which teaches thee that thou and I art one:
Shall we be sunder’d? shall we part, sweet girl?
No; let my father seek another heir.
Therefore devise with me, how we may fly,
Whither to go, and what to bear with us:
And do not seek to take your charge upon you,
To bear your griefs yourself, and leave me out;
For by this heaven, now at our sorrows pale,
Say what thou can’st, I’ll go along with thee.

Ros. Why, whither shall we go?

Cel. To seek my uncle.

Ros. Alas, what danger will it be to us,

Maids as we are, to travel so far?
Beauty provoketh thieves sooner than gold.

Cel. I’ll put myself in poor and mean attire,

And with a kind of umber smirch my face;
The like do you; so shall we pass along,
And never stir assailants.

Ros. Were it not better,

Because that I am more than common tall,
That I did suit me in all points like a man?
A boar-spear in my hand; and in my heart
Lie there what hidden woman’s fear there will,
We’ll have a swashing and a martial outside;
As many other mannish cowards have,
That do outface it with their semblances.

Cel. What shall I call thee when thou art a man?

Ros. I’ll have no other worse than Jove’s own page,

And therefore, look you, call me Ganymede.
But what will you be call’d?

Cel. Something that hath a reference to my state:

No longer Celia, but Aliena.

Ros. But, cousin, what if we assayed to steal

The clownish fool out of your father’s court?
Would he not be a comfort to our travel?

Cel. He’ll go along o’er the wide world with me;

Leave me alone to woo him: Let’s away
And get our jewels and our wealth together;
Devise the fittest time, and safest way
To hide us from pursuit that will be made
After my flight: Now go we in content,
To liberty, and not to banishment.

—From “As You Like It,” Act I.

CORYDON
By Thomas Bailey Aldrich
SCENE, A ROAD-SIDE IN ARCADY

Shepherd. Good sir, have you seen pass this way

A mischief straight from market-day?
You’d know her at a glance, I think;
Her eyes are blue, her lips are pink;
She has a way of looking back
Over her shoulder, and alack!
Who gets that look one time, good sir,
Has naught to do but follow.

Pilgrim. I have not seen this maid methinks,

Though she that passed had lips like pinks.

Shepherd. Or like two strawberries made one

By some sly trick of dew and sun.

Pilgrim. A poet.
Shepherd. Nay, a simple swain
That tends his flocks on yonder plain
Naught else I swear by book and bell.
But she that passed you marked her well
Was she not smooth as any be
That dwells here—in Arcady?

Pilgrim. Her skin was the satin bark of birches.

Shepherd. Light or dark?

Pilgrim. Quite dark.

Shepherd. Then ’twas not she.

Pilgrim. The peaches side

That next the sun is not so dyed
As was her cheek. Her hair hung down
Like summer twilight falling brown;
And when the breeze swept by, I wist
Her face was in a somber twist.

Shepherd. No that is not the maid I seek;

Her hair lies gold against her cheek,
Her yellow tresses take the morn,
Like silken tassels of the corn,
And yet brown-locks are far from bad.

Pilgrim. Now I bethink me this one had

A figure like the willow tree
Which, slight and supple, wondrously
Inclines to droop with pensive grace,
And still retain its proper place.
A foot so arched and very small
The marvel was she walked at all;
Her hand in sooth, I lack for words—
Her hand, five slender snow-white birds,
Her voice, tho’ she but said “God Speed”—
Was melody blown through a reed;
The girl Pan changed into a pipe
Had not a note so full and rife.
And then her eye—my lad, her eye!
Discreet, inviting, candid, shy,
An outward ice, an inward fire,
And lashes to the heart’s desire.
Soft fringes blacker than the sloe—

Shepherd. Good sir, which way did this one go?

Pilgrim. So he is off! The silly youth

Knoweth not love in sober sooth,
He loves—thus lads at first are blind—
No woman, only womankind.
I needs must laugh, for by the mass
No maid at all did this way pass.
 PART FOUR
Oratoric Reading and the Art of Public Speech
Discussion of forceful speech in making history. Value of forceful
speech. Practice selections.

HAMLET’S INSTRUCTION TO THE PLAYERS

Speak the speech, I pray you, as I pronounced it to you,—
trippingly on the tongue; but if you mouth it, as many of our players
do, I had as lief the town-crier spake my lines. Nor do not saw the air
too much with your hand, thus, but use all gently; for in the very
torrent, tempest, and, as I may say, whirlwind of your passion, you
must acquire and beget a temperance, that may give it smoothness.
Oh! it offends me to the soul to hear a robustious periwig-pated
fellow tear a passion to tatters,—to very rags,—to split the ears of
the groundlings; who, for the most part, are capable of nothing but
inexplicable dumb show and noise. I would have such a fellow
whipped for o’erdoing Termagant; it out-herods Herod. Pray you
avoid it.
—Shakespeare.
 CHAPTER XIII
ORATORIC READING AND THE ART OF PUBLIC SPEECH

Upon this important subject of public speaking, and the

interpretation of the addresses made by others, great men have thus
expressed themselves: Dr. Charles W. Eliot, formerly President of
Harvard University, says: “Have we not all seen, in recent years, that
leading men of business have a great need of a highly trained power
of clear and convincing expression? Business men seem to me to
need, in speech and writing, all the Roman terseness and the French
clearness. That one attainment is sufficient reward for the whole long
course of twelve years spent in liberal study.” Abraham Lincoln
likewise said: “Extemporaneous speaking should be practiced and
cultivated. It is the lawyer’s avenue to the public. However able and
faithful he may be in other respects, people are slow to bring him
business if he can not make a speech.”
Every thinker knows what a vital part eloquence plays in national
as well as individual welfare. If at first thought effective speaking
seems a simple thing and a superficial part of education, on mature
thought and consideration it will be found to be one of the most
complex, vital and difficult problems that education has to meet. And
yet, notwithstanding this complexity of the problem, the teacher is
cheered by the delightful assurance of giving the student a
consciousness of his latent talents and the ability to reveal and make
use of them for the proper influencing of his fellow men.
There is a belief fairly commonly held that only a limited few need
study the art of public speaking. Never was there a greater error or a
more fatal mistake—especially in a republic like ours, where every
man should be vitally interested in public affairs. No single citizen
can afford not to be able to stand before his fellows and clearly,
pleasingly and convincingly present his ideas upon any subject of
local, state, or national importance. It is no more an ornamental
accomplishment than is grammar, penmanship or simple arithmetic.
It should be as universal as “the three r’s.” The hints and selections
that follow are carefully chosen to incite every good citizen to the
acquirement of this useful and practical aid for his own benefit as
well as that of his fellows. All the lessons and analyses that have
gone before in these pages will materially aid in the elucidation of
these brief lessons.
The basis for development in Effective Speaking rests upon one’s
bodily, emotional and mental agencies of expression, and a
knowledge of their respective importance and efficient use. That
which counts most for development is conscientious practice; without
which, progress is impossible.
There are three definite means of communicating thought and
feeling to others: (a) Pantomime: face, hands, body; (b) Vocal: tone
sound; (c) Verbal: words, which are conventional symbols
manifesting mental and emotional states.
The problem, then, is to obtain a harmonious coördination of these
three languages. In other words, the content of the word when
spoken should be reflected in the tone and in the body. Thus speech
becomes effective merely because it receives its just and fair
consideration.
With this general understanding let us take up and master the
successive steps which ultimately lead to a realization of the desired
end.
The first important essential of effective speaking is the Spirit of
Directness. By this is meant natural, unaffected speech. Nothing can
be more important than that the person speaking use in public
address the ordinary elements of Conversation.
Hence, the first step is practice in natural speaking. Commit to
memory Hamlet’s Instructions to the Players given on a preceding
page. Do this not line by line, but the entire selection as a whole.
First: Read it through silently three times to familiarize yourself with
the subject-matter. Second: Read it aloud at least five times. Third:
Speak it conversationally at least five times from memory. In this
practice always be intensely conscious that you are addressing an
individual and not an audience.
 Now take any of the prose or poetic selections from the earlier
pages of this book, memorize them, after studying them as the
instructions require, and speak them directly and naturally, in the
ordinary conversational style.
Sufficient practice in this is the necessary preparation for the next
step, viz., the acquiring of a natural elevated conversational style,
which is merely another name for the higher type of public speaking.
Commit all, or a part, of the following selections, keeping in mind
that in speaking them you are addressing a group of people.

THE GETTYSBURG ADDRESS

By Abraham Lincoln
Fourscore and seven years ago our fathers brought forth upon this
continent a new nation, conceived in liberty, and dedicated to the
proposition that all men are created equal. Now we are engaged in a
great Civil War, testing whether that nation, or any nation, so
conceived and so dedicated, can long endure. We are met on a
great battlefield of that war. We are met to dedicate a portion of it as
the final resting place of those who here gave their lives that that
nation might live.
It is altogether fitting and proper that we should do this. But in a
larger sense we cannot dedicate, we cannot consecrate, we cannot
hallow this ground. The brave men, living and dead, who struggled
here, have consecrated it far above our power to add or detract. The
world will little note, nor long remember, what we say here, but it can
never forget what they did here.
It is for us, the living, rather to be dedicated here to the unfinished
work they have thus far so nobly carried on. It is rather for us to be
here dedicated to the great task remaining before us, that from these
honored dead we take increased devotion to the cause for which
they gave their last full measure of devotion; that we here highly
resolve that these dead shall not have died in vain, that the Union
shall, under God, have a new birth of freedom, and that the
government of the people, by the people, and for the people, shall
not perish from the earth.

By this time you should have mastered Ordinary Conversational

Style; Elevated Conversational Style; and Abandon and Flexibility of
Speech. The next consideration is the importance of Clearness.
Clearness in speech means making prominent central words and
subordinating unimportant words, or phrases. In other words, the
logical sequence of thought must be clearly shown. This is brought
about by a variety of inflections, changes of pitch, pause, etc.
Clearness in speech is dependent upon clearness of Thinking.
It is important now to give full consideration to the subject of
Emphasis. There are more ways than one of emphasizing your
thought. The most common way is by merely increasing the stress of
voice upon a word. This, however, is the most undignified form of
emphasis. It is common to ranters and “soap-box” orators and is one
mark of an undisciplined and uncultured man. Remember that
loudness is a purely physical element, and does not manifest
thought. Such emphasis is an appeal to the brute instinct, and is only
expressive of the lower emotions. But Inflection, Changes of Pitch,
Pause, Movement and Tone-Color—as have been fully explained in
preceding pages—all appeal to the exalted nature of man.
In proportion to the nobleness of an emotion or thought, we find a
tendency to accentuate these above-named elements. Such
methods of emphasis are appropriate to the most disciplined and
cultured man. More than that, they are the surest evidence of a great
personality.
Commit, then make clear to the hearer, the vital thought in the
following:

He have arbitrary power! My lords, the East India Company

have not arbitrary power to give him; the King has no arbitrary
power to give him; your Lordships have not; nor the
Commons; nor the whole legislature. We have no arbitrary
power to give, because arbitrary power is a thing which