Beruflich Dokumente
Kultur Dokumente
OF PAGES 9
Introduction
There are intrinsic limitations to the production of biofuels
derived from photosynthetic organisms that impede the
development of a renewable liquid fuel industry at largescale. In particular, these include low efficiency of solar
energy conversion (Figure 1) and competition for agricultural resources. Recent initiatives in the U.S. and elsewhere have the objective of harnessing the molecular
mechanisms of non-photosynthetic organisms that can
utilize CO2 directly for the production of energy-dense
liquid fuels, which are now referred to as electrofuels
[1,2,3]. Electrofuel-producing microorganisms are being
developed that require the complementary expertise of
www.sciencedirect.com
synthetic biologists, metabolic engineers, and microbiologists to equip native CO2-fixing species or autotrophs with
pathways for targeted fuel production, or confer autotrophy
on heterotrophic host organisms, or both. The range of
possible sources currently being explored for low-potential, high-energy electrons to power an electrofuel process
includes hydrogen gas, formate, carbon monoxide and
electricity. This review will focus on electrofuel strategies
that use hydrogen gas as source of reducing power for CO2
fixation. Microorganisms that are able to use other sources
of electrons including electricity directly are discussed in
an accompanying review by Lovely [4]. A general scheme
for electrofuel production from hydrogen and CO2 is shown
in Figure 2.
CO2 fixation
The reduction of CO2, the most oxidized form of carbon,
into technologically useful organic compounds remains a
daunting task for abiological chemical catalysis. There
are, currently, six naturally occurring biological pathways
for carbon fixation, and these have been reviewed extensively in recent years [5,6,7,8]. Each pathway has
unique features arising from the ecological and molecular
context in which it evolved. Although there are many
examples of CO2-fixing carboxylases that are utilized for
metabolic purposes other than carbon assimilation, such
as energy conservation, anaplerosis, and redox-balancing
[1,2,3,9,10], this review will focus on the autotrophic
CO2 fixation pathways that are relevant for electrofuel
production in addition to the primary host microorganisms that are currently being considered that use hydrogen (Table 1).
The most ubiquitous CO2-fixation pathway is the Calvin
BensonBassham (CBB) cycle found in plants, algae,
cyanobacteria, purple bacteria, and also in some proteobacteria, such as Ralstonia eutropha. R. eutropha is a metabolically diverse, facultatively autotrophic bacterium that
can grow on sugars, fatty acids, amino acids, triacylglycerides as well as on H2/CO2 [5,6,7,8,11]. Previous work
on R. eutropha has focused on its ability to store excess
carbon as polyhydroxyalkanoates [12,13] (PHAs) and now
efforts seek to divert carbon flux away from PHA storage
and into other molecular targets. For example, R. eutropha
is a proposed host for isobutanol production via the 2ketoisovalerate pathway for branched chain amino acid
synthesis [11] from H2 and CO2. This strategy has
already been successfully used in E. coli to produce
Current Opinion in Biotechnology 2013, 24:19
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
2 Energy biotechnology
Figure 1
(a)
CH2OH
O OH
1.8-2.4%b
0.18-0.2%c
OH
a
Wet-milling, EH
6%
4.5-
CBP
OH
OH
PS
15-2
0% d
10-13%e,f
PV
WEHP
H2/CO2
TBD
(b)
50-70%f
WEHP
70-85%f
SMR
H2/CO2
TBD
50-70%f
WEHP
Current Opinion in Biotechnology
Electrofuel production from H2 and CO2. (a) Comparison of overall photon-to-fuel efficiency of biofuels versus electrofuels. Percentages represent the
cumulative efficiency of solar energy conversion at different stages. Although the conversion efficiency of an electrofuel process is currently unknown,
the improved efficiency of solar hydrogen compared to photosynthetic sugars indicates that a solar electrofuel process would require less land area
than current biofuels. (b) Hydrogen inputs for electrofuels are highly flexible and generation strategies can include electrolysis of water by renewable
wind power, electrolysis of water by conventional electricity (e.g. coal), or by steam reformation of methane, shown with conversion efficiencies for
each process. Abbreviations: PS photosynthesis, EH enzymatic hydrolysis, PV photovoltaic, WEHP Water Electrolysis Hydrogen Production,
SMR steam methane reformation, TBD To be determined. aZhu et al. [76], bAssuming grain starch represents 40% of total corn biomass, cConrado
et al. [1], dParida et al. [77], eAssuming 65% overall electrolysis efficiency, fHolladay et al. [78].
Figure 2
+
NADP
n-butanol
NADPH
CO2
Fixation
Acetyl-CoA
Pyruvate
CO
HCO
farnesene
long chain alkenes
isobutanol
Schematic drawing of the primary biochemical modules involved in electrofuel formation. Intracellular reducing power is generated from hydrogen gas
via hydrogenase enzymes. Carbon fixation cycle incorporates inorganic carbon into central metabolism via key intermediates. Microbial synthesis of
the target fuel molecule proceeds via endogenous and/or engineered metabolic pathways.
Current Opinion in Biotechnology 2013, 24:19
www.sciencedirect.com
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
Table 1
Electrofuel producing hosts in development. The organisms listed utilize hydrogen or formate for the delivery of low-potential electrons,
and have either been reported to produce an electrofuel or are currently in development. R = reported, D = development
Organism
Type
Electron donor(s)
Betaproteobacteria
Betaproteobacteria
Archaeon
Electrochemically
generated H2
H2
H2
Clostridium ljungdahlii
Acetogenic bacteria
H2
Status
References
Target products
[18]
D
D
[11]
[3]
n-butanol, ethanol
[21]
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
4 Energy biotechnology
Hydrogen utilization
Hydrogen is used as a source of energy by microorganisms
from all three domains of life [36] and is activated by
hydrogenase, which catalyzes the reversible interconversion of molecular hydrogen and protons in the presence of
a suitable electron carrier. Based on structural and biochemical analysis, hydrogenases can be categorized by
the metal atoms in their active sites: [NiFe]-hydrogenases, [FeFe]-hydrogenases, and [Fe]-hydrogenases
[37,38]. Since electrofuel hosts containing CO2 fixation
pathways need to coordinate acquisition of reductant with
carbon flux, the types of hydrogenase present in the host
microorganism and the electron carriers that they use are
of paramount importance.
Extensive work on [NiFe]-hydrogenases from Desulfovibrio gigas [3943] and the [FeFe]-hydrogenases from
Clostridium pasteurianum [44] and Desulfovibrio desulfuricans [45] has shown that both enzyme types harbor deeply
buried active sites in which the site of H2 reactivity is a
single iron atom coordinated by diatomic ligands (carbon
www.sciencedirect.com
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
6 Energy biotechnology
Host development
The development of electrofuel hosts requires the availability of genetic tools, as well as a sound knowledge of
gene regulation and metabolism in the target host. Current electrofuel projects are taking advantage of an established body of knowledge for genetic manipulation in
well-studied organisms such as R. eutropha and Clostridium
spp., while others are working with newly developed
genetic systems (Table 2). There are two basic avenues
for electrofuel host development: engineer a natural
Table 2
Genetic tools for select electrofuels organisms. Current genetics tools for electrofuels organisms.
Organism
Genome sequence
Clostridium spp.
Many
Pyrococcus furiosus
Metallosphaera sedula
Genetic tools
Plasmid-based gene expression [62]
Gene knockout or deletion [63,65]
Chromosomal insertion [64]
Plasmid-based gene expression [68]
Anti-sense RNA silencing [68]
Expression reporter systems [68]
Chromosomal insertion [70]
Expression regulation [53]
Chromosomal insertion [27,79]
Gene knockout or deletion [72]
www.sciencedirect.com
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
Conclusion
Electrofuels are a promising paradigm that could improve
on the relatively poor efficiency of photosynthetically
produced biofuels and open up a program for scalable,
renewable liquid fuel production based on flexible energy
inputs. A handful of electrofuel organisms are in development or have been recently reported, but opportunities
exist to expand the concept. The potential of harnessing
biological carbon fixation for production of key industrial
biomolecules holds great promise for displacing
petroleum-based feedstocks [75].
Acknowledgments
This work described here was supported in part by the US Department of
Energy Research through the ARPA-E Electrofuels Program (DEAR0000081 to RMK and MWA) and the Division of Chemical Sciences,
Geosciences and Biosciences, Office of Basic Energy Sciences (DE-FG0595ER20175 to MWA).
3.
www.sciencedirect.com
5.
6.
7.
8.
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
8 Energy biotechnology
24. Estelmann S, Hugler M, Eisenreich W, Werner K, Berg IA, RamosVera WH, Say RF, Kockelkorn D, Gadon N, Fuchs G: Labeling
and enzyme studies of the central carbon metabolism in
Metallosphaera sedula. J Bacteriol 2011, 193:1191-1200.
25. Han Y, Hawkins AS, Adams MWW, Kelly RM: Epimerase (Msed_
0639) and Mutase (Msed_0638, Msed_2055) convert (S)methylmalonyl-CoA to succinyl-CoA in the Metallosphaera
sedula 3-hydroxypropionate/4-hydroxybutyrate cycle. Appl
Environ Microbiol 2012, 78:6194-6202.
26. Hawkins AS, Han Y, Bennett RK, Adams MWW, Kelly RM: Role of
4-hydroxybutyrate-CoA synthetase in the CO2 fixation cycle in
thermoacidophilic Archaea. J Biol Chem 2012 http://dx.doi.org/
10.1074/jbc.M112.413195.
43. Marques MC, Coelho R, De Lacey AL, Pereira IAC, Matias PM: The
three-dimensional structure of [NiFeSe] hydrogenase from
Desulfovibrio vulgaris Hildenborough: a hydrogenase without
a bridging ligand in the active site in its oxidised, as-isolated
state. J Mol Biol 2010, 396:893-907.
44. Peters JW, Lanzilotta WN, Lemon BJ, Seefeldt LC: X-ray crystal
structure of the Fe-only hydrogenase (CpI) from Clostridium
pasteurianum to 1.8 angstrom resolution. Science 1998,
282:1853-1858.
45. Nicolet Y, Piras C, Legrand P, Hatchikian CE, FontecillaCamps JC: Desulfovibrio desulfuricans iron hydrogenase: the
structure shows unusual coordination to an active site Fe
binuclear center. Structure 1999, 7:13-23.
46. Calusinska M, Happe T, Joris B, Wilmotte A: The surprising
diversity of clostridial hydrogenases: a comparative genomic
perspective. Microbiology 2010, 156:1575-1588.
47. Bock A, King PW, Blokesch M, Posewitz MC: Maturation of
hydrogenases. In Advances in Microbial Physiology. Edited by
Poole RK. Academic Press; 2006:1-71.
49. Burgdorf T, van der Linden E, Bernhard M, Yin QY, Back JW,
Hartog AF, Muijsers AO, de Koster CG, Albracht SPJ, Friedrich B:
The soluble NAD+-reducing [NiFe]-hydrogenase from
Ralstonia eutropha H16 consists of six subunits and can be
specifically activated by NADPH. J Bacteriol 2005,
187:3122-3132.
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017
www.sciencedirect.com
69. Lee J, Jang Y-S, Choi SJ, Im JA, Song H, Cho JH, Seung DY,
Papoutsakis ET, Bennett GN, Lee SY: Metabolic engineering of
Clostridium acetobutylicum ATCC 824 for isopropanol
butanolethanol fermentation. Appl Environ Microbiol 2012,
78:1416-1423.
70. Al-Hinai MA, Fast AG, Papoutsakis ET: Novel system for efficient
isolation of Clostridium double-crossover allelic exchange
mutants enabling markerless chromosomal gene deletions
and DNA integration. Appl Environ Microbiol 2012,
78:8112-8121.
Recent breakthrough in Clostridium genetics that opens the door for
precise metabolic engineering.
71. Heap JT, Kuehne SA, Ehsaan M, Cartman ST, Cooksley CM,
Scott JC, Minton NP: The ClosTron: mutagenesis in Clostridium
refined and streamlined. J Microbiol Methods 2010, 80:49-55.
72. Maezato Y, Johnson T, McCarthy S, Dana K, Blum P: Metal
resistance and lithoautotrophy in the extreme
thermoacidophile Metalosphaera sedula. J Bacteriol 2012,
194:6856-6863.
73. Lipscomb GL, Stirrett K, Schut GJ, Yang F, Jenney FEJ, Scott RA,
Adams MWW, Westpheling J: Natural competence in the
hyperthermophilic archaeon Pyrococcus furiosus facilitates
genetic manipulation: construction of markerless deletions of
genes encoding the two cytoplasmic hydrogenases. Appl
Environ Microbiol 2011, 77:2232-2238.
74. Bridger SL, Lancaster WA, Poole FL, Schut GJ, Adams MWW:
Genome sequencing of a genetically tractable Pyrococcus
furiosus strain reveals a highly dynamic genome. J Bacteriol
2012, 194:4097-4106.
75. Werpy T, Peterson G, Aden A, Bozell J, Holladay J, White J,
Manheim A: Top Value Added Chemicals from Biomass: Volume
I Results of Screening for Potential Candidates from Sugars and
Synthesis Gas. U.S. Department of Energy-Energy Efficiency and
Renewable Energy; 2004.
76. Zhu X-G, Long SP, Ort DR: What is the maximum efficiency with
which photosynthesis can convert solar energy into biomass?
Curr Opin Biotechnol 2008, 19:153-159.
77. Parida B, Iniyan S, Goic R: A review of solar photovoltaic
technologies. Renew Sust Energ Rev 2011, 15:1625-1636.
78. Holladay JD, Hu J, King DL, Wang Y: An overview of hydrogen
production technologies. Catal Today 2009, 139:244-260.
79. Basen M, Sun J, Adams MWW: Engineering a hyperthermophilic
archaeon for temperature-dependent product formation.
mBio 2012, 3:1-8.
Please cite this article in press as: Hawkins AS, et al.: Biological conversion of carbon dioxide and hydrogen into liquid fuels and industrial chemicals, Curr Opin Biotechnol (2013), http://dx.doi.org/
10.1016/j.copbio.2013.02.017