Algal Biotechnology: Biofuels and Beyond - Abstracts for poster presentations

Group A: Cultivation of algae for increased bioproduct

1. Environomics for scale-up of photobiological hydrogen production

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2. Evaluation of okadaic acid production by Prorocentrum lima culture using different scales of
cultivation
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3. Glycerol production by novel strains of Dunaliella and Asteromonas isolated from Namibian
marine water
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4. Chlorella emersonii cultivation, biofuel analysis and LED photobioreactor utilisation

5. Integrated production of algal biomass

Group B: Algal bioprocessing, bioprospecting and modelling for biofuel production

6. Promising catalytic processes for the chemical conversion of microalgae into biofuels

7. Biofilm of oxygenic phototrophs enhance cathode efficiency

8. Algae for cathodic oxygen reduction in MFCs

9. Micro-algal process flow-sheet energy balance optimisation: initial software evaluation

10. 'Oilgae' - A new generation of biofuel

11. Evaluation of froth flotation for recovery of algal biomass

12. Flocculation to harvest Chlorella sorokiniana

Group C: Use of algae in waste treatment

13. Microalgae and cyanobacteria in waste stabilization ponds - a molecular investigation

14. Microalgae for sustainable wastewater treatment and feed production

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15. Integration of biological wastewater treatment and algal growth for biofuels

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16. Accomplishing industrial carbon capture-an Academia for Business Project (A4B)

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17. Carbonation of an algal cultivation system

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Group D: Molecular engineering and analysis of algae

18. Exploring the potential for therapeutic protein production in algae

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19. Synthesis of antibacterial proteins in the chloroplast of the green microalga Chlamydomonas
reinhardtii
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20. Chloroplast engineering of green algae

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21. Analytical methods for research on photosynthetic organisms

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1. Environomics for scale-up of photobiological hydrogen production

P. Dechatiwongse, B. Tamburic, F. Zemichael, G. C. Maitland, K. Hellgardt
Imperial College London
Molecular hydrogen (H2) has the potential to become the fuel of the future, but only if it is produced
by a sustainable process.The green alga Chlamydomonas reinhardtii (C. reinhardtii) has an ability to
photosynthetically produce H2. It offers a biological route to renewable H2 production from the
abundant resources of sunlight and water. Photosynthetic hydrogen production is governed by the
hydrogenase enzyme, which recombines H+ + e-, to produce H2 and is activated under anaerobic
conditions. The efficient and economical growth conditions to attain healthy and high cell densities
are necessary for a commercially large-scale H2 production. As a result, key environmental
parameters e.g. light intensity, temperature and CO2 concentration have been examined to
investigate the optimum growth rate. Environomics and growth of C. reinhardtii have been
manipulated and measured using the tubular reactor, Sartorius Biostat PBR 2s.

2. Evaluation of okadaic acid production by Prorocentrum lima culture using different scales of
cultivation
R. A. Praptiwi1, C. Edwards1, L. Lawton1, C. Tang2, C. Dowle2
1

Robert Gordon University, 2The Centre for Process Innovation (CPI), Redcar

Prorocentrum lima produces several highly valuable compounds, such as okadaic acid, which are
potentially useful in medical and pharmaceutical applications. Okadaic acid (OA) is a tumour
promoter and a potent inhibitor of protein phosphatases 1 and 2A (PP1 and PP2A). OA is currently
used as an important research tool in medical and physiological studies to understand cell regulation
and various cellular processes. The aim of this study was to test the capability of P. lima in producing
OA at different scales of phototrophic cultivation.
Experiments were performed in small (1 L), medium (10 L) and large (30 L) scales of batch culture,
using P. lima PL2V strain grown in f/2 medium. Filtered natural seawater was used for the medium
preparation with salinity level of 36 ppt and pH of 6.5. Cultivation was maintained at temperature of
22 oC. Average illumination was 17.16 mol/m2/s for small and medium cultures, and 20 mol/m2/s
for large culture.
Comparison of small and medium cultures showed that optimum growth was obtained in small scale
cultivation with 3.7x105 cells/ml after 4 weeks of cultivation compared to 1.3x105 cells/ml in medium
scale culture. Analysis on the OA production also revealed that small scale culture produced much
higher OA than medium and large scale cultures, with OA concentration observed at 0.475, 0.250
and 0.195 g/ml for small, medium and large cultures respectively. This was probably caused by
difficulties in maintaining homogeneity of culture and light penetration at larger scales. Other
challenges identified from the cultivation experiment with large scale are: seawater requirement,
benthic nature of the organism, and slow growth. This indicates that optimisation of reactor design
and cultivation condition is still required for large scale cultivation of P. lima.

3. Glycerol production by novel strains of Dunaliella and Asteromonas isolated from Namibian
marine water
A.L. Abubakar (1), R. Swamy (1), J. Orchard(1), A. Highfield(2) , D. Schroeder (2), and P.J. Harvey (1)
University of Greenwich Central Avenue, (2) Marine Biological Association of the UK
Aim: to develop a low-cost system for producing industrial quantities of glycerol in pure streams
from halophytic microalgae. This study represents the first step in data acquisition for process
synthesis, modelling, optimization, and process flowsheeting aimed at integrating stages that
produce glycerol and assessing reliable targets for efficiency and cost. It centred on characterising
halophytic microalgae isolated from marine water of Namibia.
Innovation: Glycerol has emerged as a new biofuel with important properties, and now underpins a
commercial CHP technology: this is as a result of the novel Mcneil combustion cycle which allows
standard production compression ignition engines to combust glycerol at high efficiencies and with
very low emissions without chemical alteration or the addition of combustion enhancers.
Combustion is more energy efficient than any known fossil, bio or synthetic fuel (34 37 % (10
30kW), 40 42% (up to 1 MW) and 42 48% (over 1 MW) electrical efficiency), and engine
performance with glycerol has been proven. Furthermore when combusted within diesel engines
glycerol produces no combustion particulate, no SOx, reduced primary NOx, extremely low VOC and
aldehyde emissions and no catalyst poisons; these parameters have been independently verified.
Glycerol also has safe handling properties: it is water-soluble and bio-degradable and could
therefore be used to power ships and even tankers carrying glycerol as a distributable fuel with
minimal risk to the marine environment. It holds the key to developing an entirely new
environmentally-sustainable, biofuel industry with enormous commercial potential in all fields of
application of engine power production. Moreover in the biorefineries of the future glycerol will
serve additionally as an intermediate to replace various fossil oil-based bulk chemicals (e.g. ethylene
and propylene glycols, 1,4-butanediol, epichlorohydrin and acrolein).
Glycerol is currently chemically produced from biodiesel manufacture using plant oil (approximately
10% by weight of plant oil). However we anticipate that demand for glycerol will rapidly outstrip
Europes capacity to produce glycerol from plant oil. Halophytic microalgae of the Dunaliella species
that grow in highly saline environments such as salt pans and desalination non-potable waters across
the globe will also synthesise glycerol. These have been cultivated at commercial-scale as a source of
-carotene and animal protein but not as a source of glycerol, despite the fact that the basis for the
technology was established in 1978. Since the cultivation facilities depend on non-potable saline
waters, by now seeking to develop and expand halophytic microalgae cultivation for glycerol to meet
increased demand, cultivation for glycerol will help to meet EU SET targets and avoid food/fuel
competition for land use. Moreover, a very important difference between the production of algal
glycerol as opposed to algal biodiesel from lipid is that the glycerol algal product is a finished fuel
and does not require chemical modification or molecular reformation. This aspect adds benefit to
the algal production facility as glycerol derived from the plant can be used directly as a fuel to meet
the combined heat and power requirement.
Results: In this study NaCl concentration had a strong effect on the growth rate and generation time
of six strains belonging to two novel halophilic microalgae species Dunaliella (T35, T36 and T37) and
Asteromonas (T33a, T33b and T33c) as confirmed by Bayesian and Neighbour-Joining analyses. Both
species were shown to be capable of growth at different ionic strength of 0.5, to 4.0 M NaCl in a
modified Johnsons culture medium; at 12:12 photoperiod, pH 7.5, average of 4395Wm 2
photosynthetically active irradiation (PAR) and 23 0.2oC temperature. On exposure to
hyperosmotic shock the amount of glycerol produced as a function of cell density greatly increased
and was dependent on the applied NaCl concentration. After 28 days growth, for Dunaliella species
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the highest glycerol accumulation on a cell basis was obtained from T35 (203.2 3.1 pg/cell) at 4.0
M NaCl, from T36 (177.0 2.6 pg/cell) at 3.0 M NaCl and T37 (234.9 10.0 pg/cell) at 4.0 M NaCl
concentration. Ranges of 50 to 400 pg/cell glycerol have been reported by Ben-Amotz and Grunwald
(1981) for Dunaliella grown at 0.5 to 4.5 M NaCl and 94.26 pg/cell glycerol for Dunaliella species
from a hypersaline river of India (Phadwal and Signh (2003)). Similar to Dunaliella cells, Asteromonas
accumulated 53.8 0.8 to 209.5 10.1 pg/cell in modified Johnsons growth media at 1.0, 2.0, 3.0
and 4.0 M NaCl concentrations respectively. Following hyperosmotic shock glycerol accumulated
both with and without illumination, indicating that the precursor for glycerol synthesis during
hyperosmotic stress is also derived from sources other than photosynthesis, most probably from
starch breakdown.
Conclusion: The first step towards data acquisition for process flowsheeting has been achieved using
halophytic microalgae isolated from marine water of Namibia. The data provide a strategy for
cultivation scale-up to achieve large-scale glycerol production in highly saline non-potable water.

4. Chlorella emersonii cultivation, biofuel analysis and LED photobioreactor utilisation

Kirsty R. Mokebo, Prof. M.G. Davidson, Dr. L. Torrente-Murciano
University of Bath
Innovative and sustainable replacements are required for the rapidly depleting supplies of
petroleum worldwide. Biodiesel integration during the decline in use of petroleum is successfully
proven, with the infrastructure of the fuel supply network and diesel engines requiring minimal
adaptation. Algae have been cultivated using various methods, both simple and more complex.
Through the design, construction and utilisation of a specialised photobioreactor algae species can
be screened for lipid, carbohydrate and protein composition as well as for higher value products.
Gas chromatography-mass spectrometry analysis has been developed for the resultant biofuel,
generally chosen to be fatty acid methyl esters. Different growth conditions of algae, particularly the
species, Chlorella emersonii, have been investigated. To conclude, each species responds uniquely to
varying growth conditions and algal growth is furthermore affected by maturity upon inoculation
and harvesting of cultures.

5. Integrated production of algal biomass

Marco Lizzul, Dr Luiza Campos, Dr Saul Purton, Dr Frank Baganz and Paul Hellier
University College London
The burden placed on natural resources by increasing human activity has fostered a concerted drive
to redefine industrial problems within a sustainable context. This transition to a greener economy
has been recognised as vitally important to UK industry and is highlighted as key government policy.
A considerable part of this effort has been focused on the development of renewable technologies
to assist in replacing the array of chemicals derived from conventional petrochemical refineries. In
this respect, the expanding field of green biotechnology offers great promise in providing cleaner
routes to compounds made by traditional chemical synthesis. This wide-ranging potential for
biomass and its associated products has been described as the bio-refinery concept. Within this
expanding field, recent developments in algal cultivation have been identified as having the
potential to play an important role in sustainable biomass generation.

One of the major problems facing global algal production is the necessity of lowering biomass
production costs in order to make the process more economically feasible. This can be achieved in a
number of ways, but one of the most promising approaches is to integrate algal production within
existing industry and infrastructure. This approach could prove particularly effective if coupling
biomass generation to environmental remediation. Such a process would allow for the utilisation of
low cost or even completely free feedstocks, which are normally considered as waste by other
industries. Using sustainable biotechnology in this way can find parallels in the approaches
employed by systems thinkers when describing industrial symbiosis, a concept in which waste
products from one industry become the feedstock for another. With these considerations in mind,
this research project will investigate the potential benefits that can be achieved by developing more
sustainable methods of algal production, including the simultaneous design of both photobioreactor
and bioprocessing loop.

6. Promising catalytic processes for the chemical conversion of microalgae into biofuels
Li Li, Louise F. Parkes, Jack S. Rowbotham, Philip W. Dyer and H. Chris Greenwell
Durham University
The need to develop sustainable fuel sources that do not compete for land with food crops is of
growing interest. Microalgae represent possible feedstocks for achieving this aim. The methods for
converting algae into biofuels are numerous and integrating catalysts into these processes will help
to optimise their efficiency and commercial appeal. This poster aims to give an overview of some
promising processes and show how catalysts can be used to improve them.
(1) Trans-esterification is a reaction occurring between an alcohol and an ester of a second alcohol
to interconvert the functionality. For biofuel production, this typically occurs between triglycerides
(TGs) or complex fatty acids and either methanol or ethanol, to produce fatty acid methyl esters
(FAMEs). These reactions are catalysed by layered double hydroxide (LDH) materials, which are
optimised initially using simple esters, in preparation for use with microalgae.
(2) Decarboxylation is a method of chemically upgrading crude bio-oils via deoxygenation through
the release of carbon dioxide. For example, trans-esterification produces bio-diesel (composed of
FAMEs), which is oxygen rich compared to fossil fuels, therefore of limited utility. Decarboxylation
can be used to upgrade the bio-diesel to obtain a more valuable product; green diesel.
(3) Thermochemical degradation incorporates techniques such as pyrolysis and hydrothermal
liquefaction and differs substantially from conventional chemical methods. It relies on high
temperatures and heating rates to decompose long biomolecules into shorter, less oxygenated
hydrocarbons which can be burned as fuels or used to produce commodity chemicals.
Recent results exploring the efficiency of catalysts in these processes are reported, in addition to
speculation on the work that still needs to be undertaken to further enhance the efficiency of
microalgal processing.

7. Biofilm of oxygenic phototrophs enhance cathode efficiency

X. A. Waltera, I. Ieropoulosa, J. Greenmanb, C. Melhuisha
a

Universities of Bristol and of the West of England, b University of the West of England

Microbial fuel cells (MFC) are electroactive systems composed of an anode and a cathode. In such
systems, anaerobic electroactive microorganisms use the anode as an electron acceptor when
mineralising organic matter. These electrons pass through a circuit before arriving in the cathode
where they reduce a compound of a higher redox potential, thus, producing current. Typically, in
MFCs oxygen or ferricyanide accept the incoming electrons at the cathode. Power outputs are, thus,
limited by the high overpotential of oxygen. To overcome such potential, mediators or catalysts are
usually required. However, such abiotic cathodes increase the cost and lower their sustainability
through time. For this reason biocathodes have received great interest in recent years as they can
increase the power output at a lower cost and with a higher sustainability: biocathodes utilise
microorganisms as biocatalyst to mediate the reduction of an oxidant either directly or indirectly.
One of the numerous possibilities is the use of phototrophs.
The objective of the present study was to grow onto the cathode a biofilm composed of a mixture of
oxygenic phototrophs. The aim was to increase cathode efficacy by directly producing the oxygen
where it was consumed, thus, avoiding the use of any mediators. Indeed, an oxygen-supersaturation
is observed in stratified ecosystems such as in lake summer stratification or as in microbial mats.
Therefore, reproducing it in direct vicinity of the cathode would enhance the MFC output. The
stability of this current output was also investigated in order to see if such a system could be useful
for MFC applications where oxygen is a limiting factor. Figure 1 shows the relationship between
power output and light/dark conditions.

Figure 1: Mean power (n=8) output response

from a phototrophic MFC under light/dark
conditions. Error bars are present but are
smaller than the points.

The presented results confirmed that 1) the supersaturation effects of an oxygenic biofilm growing
on the cathode enhanced the current produced (Fig. 1), and 2) that this effect was stable over time.

8. Algae for cathodic oxygen reduction in MFCs

Iwona Gajda, John Greenman, Chris Melhuish, Ioannis Ieropoulos
University of the West of England
Introduction: Microbial Fuel Cells (MFCs) are systems that use microorganisms to extract energy
from organic waste. One of the MFCs major limitations is the cathodic reduction rate of oxygen
which requires an efficient catalyst. Excluding non sustainable catalysts such as ferricyanide or
expensive platinum and adapting cost effective and environmentally friendly materials, a biocathode
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is a promising alternative (1). In this study a phototrophic biocathode was developed to provide
oxygen in situ for the reduction reaction.
Materials and Methods: Five types of two chamber-MFCs were used in triplicates: 3 control abiotic
cathode MFCs and 12 biocathode MFCs separated in 4 experimental groups. The difference between
them was the cathode electrode used: a) carbon veil, b) carbon veil wrapped in cotton string, c)
carbon veil wrapped in cellulose, d) carbon veil wrapped in stainless steel wire. These electrode
configurations were used to promote algal colonisation/current collection. Cathode and anode
chamber were of equal volume and electrode surface area: 25 ml and 270cm respectively. Anode
was operated in batch mode with periodic feeding with sludge and 0.1M acetate. Cathodes were
operated in continuous flow mode connected to peristaltic pump and 0.5L photoreactors.
Results: Figure 1 shows the power curves of 4 tested biotic (a,b,c,d) and one abiotic MFC (veil
water). The maximum power density generated by the best performing biotic MFC (veil algae) was
2.78 mW/m which was 79% higher than the maximum performance of abiotic MFC that generated
1.52 mW/m.

Power density veil water

Power density [mW/m ]

3
Power density veil algae

Power density algae string

Power density algae cellulose
Power density algae wire

0
0

5
10
15
Current density mA/m

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Figure 1. Power output comparison from the different cathodic systems.

Conclusions: In the present study, algal based biocathode MFCs outperformed the Microbial fuel
cells with the abiotic cathode both in terms of power output and open circuit voltage.
References: 1. He, Z. and Angenent, L. (2006), Application of Bacterial Biocathodes in Microbial Fuel
Cells. Electroanalysis, 18: 20092015. doi: 10.1002/elan.20060362

9. Micro-algal process flow-sheet energy balance optimisation: initial software evaluation

John J. Milledge & Sonia Heaven
University of Southampton
The FP7 All-Gas project aims to produce biogas from the anaerobic digestion of micro-algal biomass
grown in open high rate ponds using nutrients from wastewater. Carbon dioxide is supplied from the
combustion of local agricultural biomass with a proportion provided from the combustion of
recycled algal biogas. The dilute nature of the algal suspension poses considerable challenges in
achieving favourable energy balances for the unit operations of algal biofuel production and
optimising the overall process.

The ability of Life Cycle Assessment (LCA) software to produce dynamic energy and mass balance
models of algal biofuel production is reviewed. An initial assessment of flow-sheet software
(Superpro, UNISIM and Aspen Plus) was carried out to assess its ability to produce a dynamic model
using parts of the possible FP7 All-Gas algal biofuel production process. It is concluded that although
UNISIM and Aspen Plus may be capable of producing dynamic mass and energy balances for an
entire algal biofuel production process it will be innovative and challenging. A very considerable
amount of effort and time will be needed to produce algal biofuel dynamic models as many unit
operations and components are not among the defaults available in either UNISIM or Aspen Plus.
Work is now examining the use of pinch analysis for not only optimisation of water usage but also on
the possibility of the use of a novel carbon and nutrient pinch.

10. 'Oilgae' - a new generation of biofuel

Holly D. Smith-Baedorf, Prof. R.J. Scott, Prof. M.G. Davidson
University of Bath
The finite nature of petrochemical energy and global climate change are driving the need for
sustainable fuels that can be readily implemented into our existing infrastructure. One such solution
may be microalgal biodiesel. With high levels of productivity, fast growth rates and few nutrient
requirements, these versatile organisms are an attractive option with the potential for many
additional applications. The project focuses on the following obstacles to realisation of microalgal
biodiesel:
1 - IDENTIFICATION OF CANDIDATE ALGAE SPECIES. As a diverse and relatively understudied group
of organisms, the isolation of temperature tolerant, filamentous or flocculating strains may reduce
cooling and harvesting costs significantly. As the only naturally 'hot' spring in the UK we are
bioprospecting the roman baths for interesting species.
2 EXTRACTABILITY. Harsh and unpredicatable environments have led to the majority of microalgae
possessing a very tough cell wall. This coupled with their small size makes the extraction of valuable
products technically difficult, usually inefficient and expensive. The project is currently developing a
screening method for the selection of cell wall mutants using staining and FACS.

11. Evaluation of froth flotation for recovery of algal biomass

Atta Ajayebi, Steve Wilkinson
University of Sheffield
In recent years, economic, environmental and energy security issues justified an investigation for
new sources of energy to replace fossil fuels. Biofuels are essentially considered as promising
substitutes for the current 80% share of fossil fuels (mainly petroleum) in the energy market.
Fundamental limitations of biofuels such as lack of arable land and fresh water encourage
researchers to look for new generations of biofuels. Algae fuel as one of these new generations has
been studied in the past decades, but high energy and production costs have been a great obstacle
against economic and sustainable production of algae fuel. In order to tackle these problems, efforts
have been made to decrease the overall costs of algae fuel production, especially the separation of
algae biomass from its growing medium, which is called harvesting of algae. In this poster a novel
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harvesting method has been proposed and studied, based on a combination of the existing air
flotation method, and froth separation, a method used in mineral processing for separation of ores.

12. Flocculation to harvest Chlorella sorokiniana

Yanan Xu, Saul Purton and Frank Baganz
University College London
Owing to the difficulties of algae biomass harvesting, flocculation is so far the most feasible
harvesting method for large scale algal biodiesel production with relatively low costs. However, most
effective chemical flocculants such as aluminum sulphate (Al2(SO4)3) will cause a problem that
recovered biomass would contain large amounts of chemical precipitation formed by the flocculants.
Chitosan is a kind of cationic polymer which can induce algal flocculation mainly due to partial charge
neutralization. It was proved highly effective in harvesting the green alga Chlorella sorokiniana at a
working pH under 7. The flocculation process was monitored with the light scattering method and
the floc size was found to grow to as large as several millimeters in a very short time. Flocculation
can significantly improve the performance of sequential dewatering processes like centrifugation and
filtration and meanwhile save much of the energy input.

13. Microalgae and cyanobacteria in waste stabilization ponds - A molecular investigation

Lucy E. Eland, Cesar R. Mota, Russell Davenport & Charlotte Paterson
Newcastle University
Waste Stabilization Ponds (WSPs) are a low cost, low energy wastewater treatment method
predominantly used in developing countries. Systems usually comprise of an anaerobic pond, a
facultative pond and a series of maturation ponds. The facultative and maturation ponds are
populated by eukaryotic microalgae and cyanobacteria that photosynthesise providing oxygen for
aerobic bacteria to treat the wastewater. The algae have also been shown to take up nutrients,
removing them from the water column.
The photosynthetic component of the biomass in WSPs has not been investigated extensively.
Molecular techniques have been used for bacterial communities and marine algal communities. This
research focuses on adapting and testing molecular techniques to assess diversity of photosynthetic
organisms in WSPs.
DNA extraction methods were tested on WSP samples, to find one that is effective and as bias free
as possible. Commercially available kits were tested for DNA quality, DNA quantity, use for
downstream amplification and diversity. Qiagens Blood and Tissue Kit outperformed the other kits
tested and does not require any specialised laboratory equipment, making it ideal for use in
developing countries.
WSP samples have been collected in two systems in Brazil for molecular analysis, one treating
domestic wastewater from homes local to the plant, the other treating a 50:50 mix of industrial and
domestic wastewater. Community analysis targeting RuBisCO genes and 18S rRNA genes aims to
investigate the effects of the waste stream on the organisms present. The study also looks at the
changes in community composition and succession seen throughout the ponds in the system.

The effects of engineering solutions on WSPs are being investigated in a pilot scale WSP in Colombia.
The effect of baffles that increase water flow and oxygen content on the community composition
and the expression of photosynthetic genes will also be assessed.

14. Microalgae for sustainable wastewater treatment and feed production

Philippe Mozzanega1, Dan Murray1, Tom Arnot1, Matthew Davidson2, Jerome Vaughan4 and Rod
Scott1
University of Bath1, Aragreen (UK) Ltd2.
Pivotal to the basic needs of modern society are access to clean water, food and fuel. Given the
added pressure of reducing our carbon intensive lifestyles and processes in order to limit potential
effects on climate change, we are exploring the viability of developing a commercial wastewater
treatment system also capable of producing biologically-derived added value products.
The removal or polishing of nitrates and phosphates from wastewater in sewage works is an energy
intensive process, which relies heavily on chemical inputs, and produces CO2, sludge and other
waste. Microalgae grown in photobioreactors offer significant potential to reduce the cost and
impact of wastewater treatment whilst contributing to the security of liquid fuel supply. In addition,
algae require large amounts of CO2 to produce biomass thereby participating in the process of
reducing global carbon emissions. Nitrates, phosphates, together with CO2 and light are the main
feedstocks for efficient algal biomass production.
Candidate species of freshwater microalgae are being assessed for their water polishing capabilities
as well as for valuable bio-products (oils, proteins (feed, food) or pigments).
A photobioreactor pilot plant has been built, which is linked to a local treatment works -operated by
Welsh Water- to provide wastewater. The pilot is now testing state-of-the-art low-energy harvesting
technologies to recover algal biomass and produce water that meets strict EU discharge standards,
while continued efforts in the lab are also looking at ways of reducing the overall energy input
(lighting, processing etc.)

15. Integration of biological wastewater treatment and algal growth for biofuels
Philippa J. Uttley
University of Sheffield
Biological wastewater treatment is an established process that harnesses microorganisms to reduce
the contaminants in domestic and industrial effluents to acceptable levels for discharge or further
processing. Photosynthetic microalgae are actively under consideration for large scale biodiesel
production since, under the right conditions, some species produce high levels of lipid that can be
separated and converted to biodiesel. This research seeks to investigate the benefits of combining
the two complementary processes via the construction of a mathematical model. By coupling our
computational model for integrated wastewater treatment and algal biodiesel production to a
mathematical optimisation algorithm, we can investigate the economics of large scale production of
microalgae using wastewater-derived nutrients, as well as enhanced production of biogas from
anaerobic digestion of algae and activated sludge. A proof-of-concept model of an integrated
WWTP has been built using simplified models of activated sludge and algal growth, utilising Monod
kinetics. Initial results illustrate the benefits of mass transfer of gases between the separate units,
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showing an increase in COD removal and an increased rate of algal biomass production. Future
versions will include anaerobic digestion and combined heat and power as producers of carbon
dioxide. The final model will employ a mathematical optimisation algorithm to find cost-optimal
designs for industrial-scale WWTPs.

16. Accomplishing industrial carbon capture-an Academia for Business Project (A4B)
Graham Nelson, Alla Silkina, Robin Shields
Swansea University
Wales carbon emissions per person are highest in the UK due to the Wales heavy industry. The 30
month project, known as ACCOMPLISH (AlgalCarbon Capture and BiOMass Production LInked Supply cHain) is being supported with more than 425,000 by EU structural funds, the Welsh
Government and resources from industrial partners, and aims to harness the properties of
microalgae to mitigate carbon emissions and provide sustainable energy.
The three main tasks include:
Development, installation and commissioning of an efficient, microalgae production system at
Tata Steel Strip Products Ltd (TATA). This system will be set up to be applied to a range of
different gas sources, including industrial steel works flue gases as well as other waste nutrient
sources.
Development of efficient processes for harvesting and drying algal biomass with the industrial
partner Axium Process Ldt through the deployment of filtration equipment.
Assessing the suitability of wet and dried micro algal biomass for bioenergy production with a
third industrial partner, using Anaerobic Digestion technology. This task is to include analyses of
moisture content, calorific and nutritional values to evaluate algal biomass as a candidate
sustainable feedstock to be integrated into existing industrial processes.
The microalgae production system developed in the ACCOMPLISH project will supply a unique
approach to provide a feasibility of steel flue gas mitigation by microalgae to reduce the carbon
footprint and assure sustainable bioenergy production.

17. Carbonation of an algal cultivation system

William Nock
University of Southampton
Cultivating microalgae for the production of biofuels is currently uneconomical in comparison to
fossil fuels. There is an on-going effort to improve the biological processes and engineering designs
to make algal biofuels economically competitive. This poster illustrates one of these concepts, which
is looking to understand and improve the supply of waste carbon dioxide into an algal cultivation
system. As well as improving the algal yield by supplying carbon to the algal cells, these designs can
also improve the de-oxygenation and mixing of the algal medium, which are both vital in improving
the effectiveness of algal cultivation systems.

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Through experimental work and numerical modeling this project has started looking into the mass
transfer from carbon dioxide bubbles into water and the effects the different chemistry of algal
media will have on this. The mass transfer of dissolved oxygen out of an algal medium has been
proven to be very important for effective algal growth and this will also be studied to better
understand the dynamics of this process. From this work it is hoped alternative designs could be
developed to maximise mass transfer in an algal media, while also enhancing the mixing that the
bubbling process generates.

18. Exploring the potential for therapeutic protein production in algae

Stephanie Braun Galleani, Frank Baganz & Saul Purton
University College London
Even though algae have been used for centuries especially as a food source, algal biotechnology is
quite novel and still remains as a non-transgenic field in commercial processes, offering much space
for development and research. Chlamydomonas reinhardtii is a well-studied organism that has been
extensively used in the past two decades for developing genetic manipulations on algae. This current
situation and the urgency to generate commercially-feasible platforms for massive recombinant
protein production have put algae in a privileged and promising position.
This research aims at analysing different factors limiting recombinant protein production in
Chlamydomonas reinhardtii by using both genetic manipulations in order to improve cell features
and protein expression, and bioprocess optimisation, studying different culture parameters that may
affect growth and protein productivity. By merging both approaches it is expected to develop an
improved platform with recombinant protein productivity comparable to current bacterial systems.
The first approach considers the incorporation an additional light-capture system (proteorhodopsin)
into Chlamydomonas reinhardtii aiming at increasing the ATP production in the cell, ongoing work
expects to conclude if this genetic modification gives some benefit on extending the culture lifespan
under nutrient starvation. A second approach considers the incorporation of a green fluorescent
protein as a marker in order to have a tool for monitoring the optimisation of culture conditions, and
it is expected to use this strain in lab scale cultivation in the coming months.

19. Synthesis of antibacterial proteins in the chloroplast of the green microalga Chlamydomonas
reinhardtii
Laura Stoffels, Bambos Charalambous & Saul Purton
University College London
Endolysins are antibacterial proteins that are produced by bacteriophages to digest the bacterial cell
wall for phage progeny release at the end of the lytic cycle. These efficient enzymes are highly
specific for the cell wall of the target bacteria without affecting other species. Development of
resistance against endolysins is very rare, because they evolved to target molecules in the cell wall
that are essential for bacterial viability. Taken together, this makes them promising novel
antibacterial agents. C. reinhardtii offers already established techniques for the expression of foreign
genes in the chloroplast and is an attractive expression platform for therapeutic proteins, due to the
lack of endotoxins and potentially infectious agents. Furthermore it can be inexpensively cultivated
in full containment and under sterile conditions in simple photobioreactors. Two bacteriophage
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endolysins targeting two major human pathogens were successfully expressed in the chloroplast of
C. reinhardtii, and the activity in killing the bacteria was assayed in vitro.

20. Chloroplast engineering of green algae

Joanna Szaub, Thanyanun Ninlayarn and Saul Purton
University College London
Our group has created an efficient method for chloroplast transformation of the green alga
Chlamydomonas reinhardtii that involves a simple procedure of agitating a DNA/cell suspension with
glass beads, and also avoids the use of any bacterial antibiotic-resistance marker. This method is
currently being applied in our research aimed at producing novel biofuel compounds or
pharmaceutical proteins in the chloroplast. For example, we have introduced a synthetic, codonoptimised gene encoding a vaccine candidate for Human Papilloma Virus, and two genes from a
cyanobacterium that encode the aldehyde decarbonylase and acyl reductase enzymes necessary for
alkane production. We are currently testing the transformed cell lines for synthesis of the
recombinant products and exploring the influence of different 5UTR and promoter elements on the
yield of the recombinant products.
Although chloroplast transformation of green algae is currently limited exclusively to
Chlamydomonas reinhardtii, our research has shown that a strain of Chlorella might make a superior
host for such recombinant production since it has significantly higher growth rate and biomass
productivity than Chlamydomonas. In order to exploit this algas potential as a platform for the
synthesis of valuable recombinant products we are developing chloroplast (and also nuclear)
transformation vectors, and testing different DNA delivery and selection methods. Our progress on
the chloroplast engineering of both Chlamydomonas and Chlorella will be presented.

21. Analytical methods for research on photosynthetic organisms

Kenneth Schneider*a, Huaining Hua, Guy Denaultb, Frank Markena and Petra J. Camerona
a

University of Bath, bUniversity of Southampton

Whether it is biofuel production, bio-electrochemical systems or fundamental biological research,

analytical methods are needed to measure quantities, which enable us to compare versatile subjects
of study and evaluate the impact of the directed change of certain factors. A couple of analytical
techniques have proven to be very useful for the selective determination of these factors in
biological investigations. The techniques presented on this poster are electro-chemical methods,
involving rotating disk electrochemistry and the use of ultramicroelectrodes as well as continuous
flow measurements. Photographs and schematics of the experimental setup are shown on the
poster, the respective advantages of the method are given and examples of the obtained
information are presented. Since most biological studies involve liquid phase studies,
electrochemistry opens up the opportunity to conduct experiments in the natural environment of
the organism. Especially the presented use of ultramicroelectrodes enables non-invasive
observations on a micrometer scale. The possibility to tune electrochemistry into high selectivity
results in a paramount sensitivity, which allows one to detect the slightest concentration changes of
the targeted species. Additionally it holds the possibility to be directly coupled with other extremely
sensitive detection methods, such as mass spectroscopy (MS) and high pressure liquid
chromatography (HPLC) for metabolic profiling and flow cytometry for growth rate monitoring.
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