Tredici Photobiology 2010

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Review
Photobiology of microalgae mass cultures: understanding

the tools for the next green revolution
Biofuels (2010) 1(1), 143–162
Mario R Tredici†
Several obstacles and limitations currently prevent the industrial exploitation of microalgae for feed, food
and biofuel production. Photobioreactors (closed systems for algae cultivation) suffer from high‑energy
expenditures for mixing and cooling, while cultures in large-scale open ponds, which have a more
favorable net energy ratio, are unstable ecosystems in which maintaining selected strains for long periods
is difficult. Techniques for supplying nutrients and CO2, for mixing and for harvesting and processing the
biomass in an energy-efficient manner are still under study and development. Despite these impediments
and although microalgae are not superior to higher plants in terms of photosynthetic efficiency and
productivity, microalgal cultures remain one of the most attractive sources of feed, food and next-generation
biofuels since microalgae can be grown in saline or seawater on nonarable lands, can use fertilizers with
an almost 100% efficiency, are able to attain much higher oil and protein yields than traditional crops and,
being endowed with high growth rates, are easier to be improved via genetic and metabolic engineering.
Microbial oxygenic phototrophs (microalgae and cyano composition by varying culture conditions [1] . They can
bacteria) are believed to be very efficient solar energy con be grown under nutrient limitation and thus allow the
verters and for this reason they are considered attractive for use of fertilizers with almost 100% efficiency. Microalgae
renewable biofuel production [1,2] . However, in contrast mass cultivation (with suitable strains and technologies)
to common belief, microalgal cultures are not superior to does not necessarily rely on chemical molecules to control
higher plants in terms of photosynthetic efficiency and pathogens and predators, and can be carried out in saline
productivity. Microalgae present several advantages over or seawater on nonarable lands. This last characteristic
plants as sources of second-generation biofuels, but they may become critical when agro-industry will choose the
neither achieve higher solar radiation capture and conver next generation of crops to tackle the dramatic problem
sion efficiencies nor do they possess better mechanisms of increasing food production under decreasing resource
for photoprotection. Microalgae lack heavy supporting availability (e.g., fossil fuels, fertile soils and water) and
structures and anchorage organs and, different from most climate change. Last but not least, despite similar efficien
plants, microalgal cultures attain a complete land coverage cies and productivities, microalgae can attain much higher
throughout the year. However, currently, these character oil yields per hectare than traditional crops [3] .
istics do not translate into a real advantage in terms of bio The concept of ‘mariculture on land’, the large-
mass production per unit land surface area occupied. The scale cultivation of selected marine microalgae on arid
reasons of their inefficiency are illustrated in this review. coastal areas, was proposed in the late 1970s for energy
The real advantages of microalgae over plants reside in purposes [4,5] . Nowadays, humankind is faced with
their metabolic flexibility, which offers the possibility of other, more urgent, necessities owing to the increased
modification of their biochemical pathways (e.g., towards world population, climate change and impoverished
protein, carbohydrate or oil synthesis) and cellular earth’s resources. In this article, mariculture on land is
†
Author for correspondence
Dipartimento di Biotecnologie Agrarie – Università degli Studi di Firenze, Piazzale delle Cascine 24, 50144 Firenze, Italy; Tel.: +39 055 328 8306;
Fax: +39 055 328 8272; E-mail: [email protected]
future science group 10.4155/BFS.09.10 © 2010 Future Science Ltd ISSN 1759-7269 143
Review Tredici
Key terms reproposed, after careful evaluation Appropriate techniques to build large-scale ponds for
Microalgal cultures: Artificial of the potential of algal photosynthe bulk cultivation and bioreactors necessary to provide
systems where algae are grown for sis and its limitations, with a broader large amounts of inoculum;
particular applications scope: producing a novel source of
Efficient techniques for separating the microscopic
Photosynthetic efficiency: The biomass from which to derive food,
efficiency with which the energy of light cells from the culture solution and processing the
feed, renewable fuels and chemicals,
is converted into the chemical energy of biomass to obtain the desired product.
carbohydrate or biomass
using seawater, sun energy and arid or
marginal lands as the main resources. Obviously, all this has to be done cost effectively in
Photobioreactors: Closed system for
algae cultivation
Mariculture on land will have the order to compete with conventional agriculture and
possibility to succeed and reach the fossil fuels. This review analyzes the main reasons that
Algae biofuels: Biofuels derived from
algae biomass (e.g., algae oil, bioethanol market only if we recognize, today, limit the photosynthetic performance of algal cultures
or biodiesel) or attained through algal that there are several and severe in comparison with plant systems, compares perfor
metabolism (e.g., hydrogen) bottlenecks that hamper algae culti mances of photobioreactors and open ponds, and con
Oxygenic photosynthesis: Coordinated vation at large industrial scales. Algae siders the potential of microalgae for oil and protein
complex of biochemical reactions used advantages and drawbacks must be production. Marine microalgae can provide answers
by oxygenic phototrophs to convert
considered, without excessive enthu to the global energy crisis and ongoing changes to our
light energy into chemical energy. The
process releases oxygen as a by-product siasm or prejudices, but exclusively planet’s climate, since they neither need fertile soils nor
with a scientific approach. What are freshwater. To illustrate the potential of algal photo
the real advantages of microalgae synthesis, freshwater algae, on which more numerous
over plants? Where are the main limitations of large- studies have been carried out, are also considered.
scale microalgal cultures? What can we reasonably expect
to achieve in half a decade of research from now in the Reactions of oxygenic photosynthesis
algae–biofuel arena and what is beyond our reach? In the Photosynthesis literally means ‘building up by light’ [7] .
past years, much emphasis has been put on the photo In this process, energy-poor inorganic substances (CO2,
synthetic efficiency and productivity of algae and into water and mineral salts) are converted to energy-rich
photobioreactor design [6] . However, after more than 50 organic compounds and oxygen is released as a byprod
years of R&D, there is not yet an algal strain or reactor uct. Overall, the solar energy absorbed by photo
or combination of both able to achieve large-scale (100s synthetic pigments is converted into the chemical energy
of hectares) yields comparable to C4 plants (e.g., sugar of biomass. The reaction can be simply described as:
cane) and to compete with plants in producing biofuels or Reaction 1
protein. There are interesting old and new ideas that may
lead to higher photosynthetic efficiencies and productivi CO 2 + H 2 O + sunlight $ 6 CH 2 O @ + O 2 + waste heat
ties with microalgae (e.g., reduced antenna mutants, light
dilution and optical fibers), but they must be proved at where [CH 2O] represents a nominal carbohydrate
large scales and their application is, at present, cost pro (e.g., one sixth of d-glucose) molecule, the immediate
hibitive. Besides, and above all, it must be said that no end product of photosynthesis. The initial products of
technology will help to break the laws of physics. There CO2 fixation – sugars – are later converted to a vari
have been numerous recent claims of yields attained ety of secondary products, for example structural (e.g.,
or attainable with miraculous strains and bioreactors cellulose) or energy-storage polymers (e.g., starch and
that largely surpass the theoretical efficiency of photo fats) and functional molecules (e.g., enzymes, DNA,
synthesis and, in some cases, even break the first law RNA, pigments, phospholipids, different low-molecular
of thermodynamics [201] . As expected, the claims were, weight compounds), that are necessary for cell growth.
and remain, unsubstantiated. It is perhaps useful to say Photosynthesis can be also viewed as a redox reaction
that extremely high yields are not a necessary requisite in which electrons are transferred from water to CO2
to exploit microa lgae for the large-scale production of (CO2, the electron acceptor, is reduced and water, the
protein and biofuels. What we essentially need today are: electron donor, is oxidized). Since the transfer of elec
trons occurs against the electrochemical gradient, the
Robust marine algal strains and appropriate cultiva reaction is endothermic and requires a source of energy
tion technologies able to guarantee a sustained and (light) to proceed. Chlorophyll molecules are the site
relatively high biomass productivity; of the initial photochemical reactions and function as
both light absorbers and, in the excited state, as sources
Efficient methodologies to supply the algal cultures of electrons. Special chlorophylls absorb light and eject
with the required carbon and nutrient sources; strongly reducing electrons that are then transferred
144 Biofuels (2010) 1(1) future science group

Photobiology of microalgae mass cultures: understanding the tools for the next green revolution Review
Photochemistry
According to the now widely accepted ‘Z-scheme’
of photosynthesis, first proposed in 1960 [8] , two
O2 CO2 different photochemical membrane-bound systems
NADPH (containing special chlorophylls in the reaction cen
Light Glucose
(strong reductant) ter and several molecules of antenna pigments), are
H 2O involved in the electron transfer reactions from water
Dark biochemistry to NADP: photos ystem II (PSII) and photosystem I
(PSI). The two systems operate in series, with sev
Figure 1. Photosynthetic reactions. eral membrane and soluble complexes and molecules
acting as intermediate carriers (Figure 2) . Thus, two
photons are necessary (one absorbed by PSII and one
by several electron carriers to a final acceptor, NADP by PSI) to move one electron uphill from water to
(Figures 1 & 2) . The pigments are thus oxidized and NADP [9] . Since the formation of one molecule of
NADP is reduced (to NADPH). The photo-oxidized NADPH requires two electrons and consequently
chlorophyll is so reactive that a molecule of water, a four photons, to obtain the two moles of NADPH
very stable compound, is forced to split and the electron necessary to fix one mole of CO2 , a minimum of
hole in the chlorophyll molecule is replenished by water eight moles of photons is required. The absorption
oxidation. The light-driven electron transport in the of eight photons also leads to the accumulation of 12
photosynthetic membrane also generates ATP (Figure 2) , protons in the chloroplast lumen. With four protons
which, together with NADPH, is then used in the required for the synthesis of one ATP [10] , this is
so‑called dark reactions of the Calvin–Benson cycle to just sufficient to synthesize the three ATP necessary,
fix (reduce) CO2. In the cycle, used by plants, algae and together with the two NADPH, for reduction of one
photoautotrophic bacteria, 18 ATP and 12 NADPH molecule of CO2 to its sugar equivalent (CH 2O) in
molecules are required for every six molecules of CO2 the Calvin cycle (Reaction 2) . The energy content of
converted to one molecule of glucose [7] . The reaction NADPH is 220 kJ mole -1 and that of ATP is 30.7
can be represented as: kJ mole -1 [7] , thus the total energy expenditure in
Reaction 2 the fixation of CO2 (Reaction 2) is 532 kJ mole-1 CO2 .
CO2 + 2 NADPH + 3 ATP $ Since the energy stored in [CH 2O] is 477 kJ mole-1,
6CH2 O@ + 2 NADP + 3 ADP + 3 Pi
Reaction 2 has a negative DG of 55 kJ mole -1 and may
proceed spontaneously.
H+
Electron transfer pathway
ADP ATP
Photon H+ Photon
NADP NADPH
PSII Cyt b6f PSI e-
FD
FNR
QB Cyt b 6 FeS
QA PQH 2
Pheo FeS A ATP-synthase

PQ
e- e-
P680 Cyt f P700
PC
H 2O ½O2+ 2H +
H +
H+
Figure 2. The photosynthetic membrane in oxygenic phototrophs. Light is captured by light-harvesting pigment–
protein complexes and is funnelled to the reaction centers of PSII and PSI. The two photosystems cooperate to
move electrons from water to NADP. The diagram also shows how the proton gradient is generated by the oxidation
of water and of reduced plastoquinone by the Cyt b6f complex and is then used by ATP synthase to synthesize ATP.
Cyt b6 and Cytf: Cytochromes; FD: Ferredoxin; FeS: Nonheme iron-sulfur proteins; FNR: Ferredoxin-NADP reductase;
P680 and P700: Reaction centre chlorophylls of PSII and PSI; PC: Plastocyanin; Pheo: Pheophytin; PQH2: Mobile pool
of plastoquinone molecules; PSII and PSI: Photosystems; PQ: Plastoquinone; QA: Bound plastoquinone; QB: Reversibly
bound plastoquinone.
future science group www.future-science.com 145

Review Tredici
When absorbed instead of impinging light is con net photosynthesis can be measured. From the com
sidered, the number of CO2 molecules fixed (or oxygen pensation point on, P increases linearly with I up to a
evolved) per photon and the number of photons nec certain value, beyond which the slope of the PI-curve
essary to fix one CO2 molecule (or evolve one oxygen decreases progressively. If irradiance increases further,
molecule) may be defined as the quantum yield and a saturation value is reached (Is) at which P does not
the quantum requirement of the process, respectively. increase despite the increased irradiance. Here, irra
The maximum (theoretical) quantum yield will then diance is saturating and photosynthesis is said to be
be 0.125 mol CO2 fixed (or oxygen evolved) mol-1 pho light-saturated. At low irradiances, in the light-limited
tons absorbed, corresponding to a quantum requirement region, P is linearly proportional to irradiance and a
of eight. doubling of irradiance produces an almost doubling
of P; the maximum rate of photosynthesis per unit of
Light–response curve of photosynthesis incident irradiance is achieved here. The ratio between
The rate of photosynthesis of a phototrophic cell photosynthesis and irradiance in this initial portion of
depends on the rate of photon absorption and on the the curve (called the initial slope a of the PI-curve in
efficiency with which the absorbed photons are con the aquatic sciences literature), is proportional to the
verted into chemical energy by the photosynthetic maximum quantum yield of photosynthesis and gives a
reactions. Absorption depends on the absorption measure of the maximum efficiency of light conversion
cross-section of the cell and irradiance. If the rate of into biomass. The initial slope is related to the maxi
photosynthesis (P) is plotted as a function of irradi mum quantum yield of photosynthesis through light
ance (I), the so-called light–response curve of photo absorption. If the initial slope is measured in a culture
synthesis or PI-curve (Figure 3) is obtained [11,12,202] . so dense or in a leaf, that all the incident photosyn
The PI-curve can be divided into three distinct zones: thetic active radiation (PAR) is absorbed, it is approxi
the photolimited, the photosaturated and the pho mately the same as that measured for absorbed light
toinhibited region (Figure 3) . In darkness, there is no and can be taken as a direct measure of the maximum
photosynthetic activity and only oxygen consumption quantum yield of photosynthesis [13] . At the saturat
due to cellular respiration is measured. As irradiance ing irradiance and above, the system operates at the
gradually increases, a value, called the light compensa maximum permissible photosynthetic rate (Pmax) for
tion point or compensation irradiance (Ic), is reached that organism under the experimental conditions. In
at which photosynthetic oxygen production balances this light-saturated region, the rate of photon absorp
respiratory oxygen consumption. At higher irradiance tion exceeds the rate of electron transport from water
values, oxygen production exceeds consumption and to CO2 : the dark enzymatic reactions are working as
fast as they are capable and cannot make any use of
additional absorbed photons, limiting the photosyn
Photolimitation Photosaturation Photoinhibition thetic rate [10,11] . Thus, the efficiency of conversion of
Pmax light energy into chemical energy progressively falls
from the end of the linear region to the light saturated
Photosynthesis rate
region of the PI-curve.

It is also convenient to consider the interception of
a and Pmax (Figure 3) . This point, never actually mea
sured, is often called the light-saturation parameter and
denoted by Ik (Pmax /a). Ik is the irradiance at which
control passes from light absorption and photochemical
α energy conversion to reductant utilization and is used as
a convenient index of the photoacclimation status [13] .
Ic Ik Is Ih
If the irradiance increases much beyond light satu
Irradiance
ration, it may eventually reach a value (I h) at which
photoinhibition sets in and light becomes injurious.
Figure 3. PI-curve that relates photosynthesis rate to irradiance.
Consequently, in the photoinhibited region of the
Ic: Compensation irradiance, the point at which photosynthetic oxygen
PI-curve, P decreases below Pmax. Photoinhibition is
production balances respiratory oxygen consumption; Ih: Irradiance
dependent on both intensity of the light and duration of
value at which photoinhibition sets in and light become injurious; Ik: The
exposure. Since in many freshwater and marine micro
irradiance at which control passes from light absorption and photochemical
algae, irradiances in the 100–200 µmol photons m-2 s-1
energy conversion to reductant utilization; Is: Saturation value, the point at
range (less than one tenth of full sunlight) may satu
which P does not increase despite the increased irradiance.
rate or even photoinhibit, photosynthesis [10,12] , light

saturation and photoinhibition are the most serious various researchers in the past, having measured lower
obstacles to achieve high photosynthetic efficiency (PE) quantum requirements, have sustained that the com
under natural illumination in algal cultures (see later). bined action of two photosystems was not necessary to
reduce CO2. The determination of the minimum quan
Maximum (theoretical) efficiency of tum requirement of photosynthesis has been the subject
photosynthetic solar energy conversion of a bitter controversy between Otto Warburg, Nobel
Establishing the maximum (theoretical) energy-storage laureate of Physiology & Medicine, and Robert Emerson,
efficiency of photosynthesis (from charge separation in discoverer of the photosynthetic unit, the red drop and
the chlorophylls to carbohydrate) under sunlight is the Emerson enhancement effect [16] . Warburg insisted
of great interest. It fixes the maximum biomass yield that the minimum quantum requirement was three to
achievable from a given area of crop plantation or four, whereas Emerson and others consistently obtained
algal culture or, in practical terms, the minimum land values of eight to 12. The controversy lasted for more
area required to produce a given number of food or than 25 years and remains unsolved, in the sense that
fuel calories. no-one has found the reason for the large discrepancy in
The maximum efficiency of photosynthesis (MPE) the measures between these two eminent scientists. It is
can be calculated considering that the chemical energy interesting to note that, as a conclusion to the diatribe, in
stored in one mole of fixed CO2 amounts to 477 kJ his last and final paper on the topic [17] , Warburg himself
(one sixth of a mole of glucose) and, according to the published a measured minimum quantum requirement
Z-scheme, a minimum of eight red photons (171 kJ of 12, in full agreement with Emerson’s values [16] .
mole-1) are required per molecule of CO2. The MPE In the early 1980s John Pirt and co-workers measured,
attainable with red light is thus 35% (477/[171 × 8]). under artificial white light, a PE of 35% in chemostat
Shorter wavelength (higher energy) PAR photons are Chlorella cultures and of 47% with a consortium of
effective, but higher excited states of chlorophyll rapidly Chlorella and three heterotrophic bacteria [18,19] . Based
relax and the additional energy is wasted. The process is on these results, Pirt, who did not take into account the
driven with the energy of a red photon regardless of the light saturation effect (LSE; see following sections), esti
wavelength absorbed [14] . Given the average energy con mated a maximum possible photosynthetic efficiency
tent of PAR of 217 kJ mole-1 (Box 1) , the energy lost due exceeding 18% under full sunlight. These findings have
to degradation of PAR to excitation energy at 700 nm never been substantiated.
is 46 kJ (217–171 kJ) mole-1 PAR photons or 21% of
Box 1. Photosynthetically active radiation.
the absorbed PAR energy. Taking an average PAR of
45% in solar radiation (Box 1) and the 21% loss due to Only a fraction of the solar radiation spectrum has the appropriate energy
degradation of PAR to red light, the MPE achievable content to be used in photosynthesis. It is called photosynthetically active
under sunlight is 12.4% (0.35 × 0.79 × 0.45). More radiation (PAR) and is made up of photons in the wavelength range from
400 nm (blue photons) to 700 nm (red photons). In free space the energy of a
simply, this value may be calculated as the ratio between
photon or quantum of light is:
the energy of the reduced mole of CO2 (477 kJ) and the
hc
energy of eight moles of PAR photons (1736 kJ), thus E = hv =
m
obtaining a MPE on PAR of 27.5% and considering in which, E: Energy content of a single quantum (J) ; h: Planck’s constant
that PAR is 45% of total solar radiation (0.275 × 0.45). (6.626 × 10 -34 Js); n: Frequency of radiation; l = wavelength (m) and c: Speed of
The minimum quantum requirement must be sub light propagation (2.9979 × 108 m s-1).
stantially increased (and the overall efficiency decreased) According to the equation, red photons (700 nm) have an energy content
when biomass, instead of carbohydrate, is considered of 171 kJ mole -1, 43% lower than blue photons (299 kJ mole -1). Solar PAR has
as the final product of photosynthesis. Biomass also an average energy content of 217 kJ mole-1. Photons of wavelength above
contains proteins, lipids, nucleic acids and other con 700 nm do not contain sufficient energy (the so-called threshold value) to
stituents, the biosynthesis of which from carbohydrate eject electrons from chlorophyll and induce charge separation.
Light absorption is wavelength-dependent, but every PAR photon absorbed
requires additional energy and reducing power. The
by the antennae can be used in the reaction centers to induce charge
likely minimum requirement to fix one mole of CO2 into
separation with the same efficiency and results in an equal photosynthetic
biomass varies from 10 to 12 moles of photons, instead of output. Since the quantum yield is more or less constant in the PAR range (but
eight [12,15] , which decreases the MPE to less than 10%. see [11,12 ] for a decline of quantum yield below 575 nm), in energy terms the
photosynthetic efficiency regularly increases from 400 to 700 nm, because the
Oxygenic photosynthesis: minimum quantum photon energy content decreases with increasing wavelength.
requirements lower than eight The fraction of PAR in solar radiation varies from 43 to 48% depending
The Z-scheme of photosynthesis, which defines a mini on the angle of incidence and atmospheric conditions, thus more than half
mal quantum requirement of eight, is now widely recog of the solar radiation that reaches the earth is below the threshold and is
nized as valid [7–14] . It must be mentioned, however, that useless for photosynthesis.

Review Tredici
More recently a Chlamydomonas reinhardtii mutant local partial pressures of the substrates (CO2 and O2).
lacking PSI has been found to be capable of photoauto Photorespiration would be annulled at approximately
trophic assimilation of CO2 and oxygen evolution [20] , 700 ppm of CO2 in air. At the present atmospheric
suggesting again that a single-photon light reaction CO2 concentration (380 ppm) and high temperatures,
driven by PSII alone is able to generate the necessary photorespiration in some inefficient plant species may
chemical potential for reduction of CO2. Surprisingly, waste up to 50% of net photosynthesis [7,14] . Barber and
the quantum yields for the reduction of NADP were Archer report losses of fixed carbon for photorespiration
about the same for the mutant and the wild-type varying from 30% (at 25°C) to 40% (at 35°C) [9] , while
alga, and photoautotrophic growth was not observed Zhu et al. assumed a loss of 48% [14] . Here a minimum
under aerobic conditions. Considering the reservations loss of 40% is considered.
expressed by the researchers themselves [21] and others Many tropical grasses and crops (such as maize, sor
regarding whether the mutant was completely deprived ghum and sugarcane) fix CO2 into four-carbon com
of PSI [7] , we can reaffirm the validity of the Z-scheme. pounds (hence they are said to be C4 plants) before it
enters the common C3 pathway. The CO2 that diffuses
Maximum photosynthetic efficiency in the into the leaf is initially fixed onto phosphoenolpyruvate
field (plants) in the mesophyll by phosphoenolpyruvate carboxyl
Thanks to photosynthesis, sun energy is stored in a ase to form oxaloacetate and then malate or aspartate
form (chemical energy of sugar) that can be both bio [22] . The C4 dicarboxylic acids are transported into the
logically used or easily converted to fuel. Unfortunately, bundle-sheath cells, where they are decarboxylated to
photosynthesis is not a perfect process and from the pyruvic acid, which is returned to the mesophyll to
initial sugar to the final biomass in the field a number regenerate phosphoenolpyruvate and initiate another
of important losses exist. Even when all other determi cycle. The released CO2 is reduced, in the bundle-
nants of productivity (e.g., nutrient and water availabil sheath cells (apparently impermeable to CO2), via the
ity, temperature and developmental state of the plant) Calvin–Benson cycle [7,22] . Thanks to this mechanism
are at the optimum and in the absence of physical and that concentrates CO2 at the site of RuBisCo, the oxy
biological stresses, the maximum PE attainable under genase activity of the enzyme is largely prevented and
full sunlight is much below the theoretical maximum. the losses due to photorespiration are eliminated or
Losses due to reflection, photorespiration, respiration, significantly reduced [14] . Under moderate tempera
photosaturation and photoinhibition diminish the ture and irradiances and adequate water supply, the
photosynthetic efficiency to well below 12.4%. quantum yield in C3 and C4 plants is similar, but C4
plants are much advantaged in warm temperatures and
Loss by reflection under high irradiances. It is also a general opinion that
Reflection from a single leaf is angular-dependent and photosynthesis in C4 plants is not fully saturated even
can be higher than 15%, but the plant canopy reabsorbs at high irradiances [7] and that it is less sensitive to
part of the reflected and diffused light, reducing net photoinhibition. Cloudy conditions remove most of the
reflection. We can assume that the minimum fraction advantage of C4 metabolism.
of PAR reflected by plant canopies, which absorb weakly The concentration mechanism of C4 plants, how
in the green light band, is approximately 10% [14] . ever, requires an additional consumption of two ATP
molecules per molecule of fixed CO2. The eight protons
Photorespiration needed to synthesize these two additional ATP mol
CO2 fixation in all oxygenic phototrophs relies on ecules are obtained by the absorption of four additional
the activity of the enzyme ribulose-biphosphate car photons by PSI and cyclic photophosphorylation [14] .
boxylase/oxygenase (RuBisCo). In the first step of This brings the energy input to fix one mole of CO2 to
the Calvin–Benson cycle, RuBisCo catalyzes the car 2604 (12 × 217) kJ and reduces the maximum theoreti
boxylation of ribulose-1,5-biphosphate (RuBP) and cal photosynthetic efficiency in C4 plants to (477/2604)
the formation of two molecules of 3-phosphoglycer 18.3% on PAR or 8.2% on total solar radiation.
ate (hence C3 cycle) [7] . RuBisCo also catalyzes the
oxidation of RuBP (in a process commonly known as Respiration
photorespiration), which ultimately results in release of In contrast to photorespiration, normal dark respiration
CO2 and ammonia [22] . Photorespiration requires ATP is a necessity. All phototrophs respire (in darkness and in
and reducing equivalents and thus wastes considerable the light) to maintain cell integrity and to stay alive. The
amounts of fixed energy. The two reactions, carboxyl losses of fixed carbon and energy caused by respiration
ation and oxygenation, compete for the enzyme active are difficult to estimate. While losses of CO2 as high as
site, making RuBisCo activity extremely sensitive to 60% have been measured for major crops [14] , minimum

respiratory losses of 15–20% have been observed in the day, when photosynthesis is more light-limited, a large
both terrestrial and aquatic phototrophs [14,23] . Here a proportion of beam radiation is lost by reflection because
conservative minimum of 20% is considered. sunlight impinges at large angles. For use in field esti
mates, a daily average loss of 15% has been suggested [11] .
Photosaturation & photoinhibition I will here assume that a minimum of 10% of the incident
Photosaturation and photoinhibition almost universally visible radiation is reflected at the air/water surface and
affect photosynthesis [7,9,10,14] and may further reduce does not penetrate below the water surface during a typi
the aforementioned yields by a minimum of 20% in cal clear day. Absorption by nonphotosynthetically active
C3 plants and 10% in C4 plants. The reason of photo components is not considered.
saturation is essentially that the antenna pigment sys
tems of plants have been optimized for the daily average Respiration & photorespiration in microalgae
intensity at the expense of efficiency at high irradiances. During photosynthesis, decay processes (i.e., dark res
During the central daylight hours of a clear summer piration, photorespiration and cell death) are at work in
day, photons are harvested at a rate much faster than microalgal cultures. At high irradiances, with increas
the reaction centers can process them and often above ing O2 and decreasing CO2 concentrations, photo
irradiation levels of half full-sunlight there is no further respiration may become the main decay process, while
increase of photosynthesis [14] . The problem is not only at low irradiances, respiration is relatively more impor
reduction of the maximum efficiency, but it goes fur tant [11] . In natural waters, photorespiration and excre
ther: if the absorbed energy exceeds the capacity of dis tion of glycolic acid can account for a loss of 50% or
sipation mechanisms, damage occurs to the photosyn more of the photosynthetically assimilated carbon [11] .
thetic apparatus (photoinhibition). Much uncertainty The real influence of photorespiration in mass microal
still remains on the influence of photoinhibition on PE gal cultures is, however, difficult to estimate, changing
and productivity, because plants, especially sunlight- strongly with the species and the culture conditions [22] .
adapted plants, as are most of the C4, are endowed with For example, both respiration and photorespiration
powerful mechanisms to avoid excess light and cope increase with temperature, while ATP-dependent CO2-
with photosaturation and photoinhibition. concentrating mechanisms that reduce the losses by
photorespiration may be activated by some microalgae
Maximum photosynthetic efficiency in outdoor under low CO2. Here it will be assumed that in well-
algal cultures managed cultures, where oxygen is efficiently removed
For algal cultures, the overall PE can be calculated as the and CO2 supplied at sufficient rates, the O2 :CO2 ratio
ratio between the chemical energy stored in the biomass at RuBisCo’s active site is low enough so as to practi
(i.e., the heat of combustion of the biomass produced cally eliminate photorespiration. Differently, respira
per surface area per time) and the irradiance (in energy tion cannot be ignored and a minimum loss of 20%
terms) at the culture surface. A heat of combustion of of the fixed carbon will be considered, although night
20–23 kJ g-1 (dry wt) is typical for algae grown under respiratory losses up to 40–50% of the carbon fixed in
nutrient-sufficient conditions and with an average lipid the light have been measured. Night respiration losses
content of approximately 20% [11,23] . depend on the temperature and the light level under
The mechanisms of photosynthesis described earlier which the culture has been grown. Exudation of organic
(Figures 1 & 2) and PAR utilization are common features material and production of exopolysaccharide, common
to all oxygenic phototrophs (including prokaryotes), phenomena in diatoms and cyanobacteria that further
thus the value of 12.4% (maximum for C3 metabo reduce yields, are not considered.
lism) can be taken as the MPE of conversion of solar
light by algae (micro- and macro-algae and cyanobacte Light saturation effect in outdoor algal cultures
ria) cultures as well, before reflection, photorespiration, With zero losses for photorespiration, mass algal cul
respiration, photosaturation and photoinhibition losses. tures would end up with a significantly higher PE in the
field than plants (Table 1) . Unfortunately, these values
Loss of light by reflection in water bodies are much reduced in algae mass cultures exposed to full
In water bodies, reflectance is mainly influenced by the sunlight. The problem is high irradiance: too much of
angle of incidence of the sun rays and by the smoothness a good thing [202] . While efficiencies of up to 24% of
of the water surface (which is modified by wind or mix PAR (11% of total solar radiation) have been measured
ing). While only a small percentage is reflected on a calm with several microalgae under low-light conditions [24] ,
clear day with the sun just ahead, more than 30% of the even under illumination of approximately half of the
incident light may be lost with strong winds and large sunlight intensity, the efficiency decreases to less than
angles [12] . It is of note that in the early and late hours of 7% of PAR [25] .

Review Tredici
Table 1. Minimum energy losses of total incident solar radiation in C3 and C4 plants and microalgae, from
interception to formation of carbohydrate.
Minimum energy losses Energy remaining (%)
C3 plants C4 plants Microalgae mass cultures
Total incident solar radiation 100 100 100
Radiation outside PAR (55%) 45 45 45
Degradation of absorbed PAR photons to excitation energy at 35.6 35.6 35.6
700 nm (21%)
Conversion of excitation energy at 700 nm to the chemical energy 12.4 (MPE) 8.2 (MPE) 12.4 (MPE)
of glucose (65% in microalgae and C3 plants; 77% in C4 plants)
Reflection (10%) 11.2 7.4 11.2
Photorespiration (40% in C3 plants, none in C4 plants 6.7 7.4 11.2
and microalgae)
Respiration (20%) 5.4 5.9 9.0
Photosaturation and photoinhibition (20% in C3 plants; 10% in 4.3 5.3 5.4
C4 plants; 40% in microalgae)
MPE: Maximum efficiency of photosynthesis; PAR: Photosynthetically active radiation.
The problem is that in dense cultures, the cells spend pigment complexes (see following sections). This causes a
most of their time in the poorly lit layers and consequently sharp attenuation of light along the culture depth (Figure 4)
acclimate to low light by assembling very large antenna and the formation of layers of different irradiance to which
the cells are exposed in a very complex cycle depending
2000 on culture depth, cell concentration and mixing rate
0.3 g/l (Box 2 & Figure 5) .
1800 1 g/l Whether acclimation to lower irradiances involves
2 g/l larger photosynthetic unit (PSU) size (i.e., a higher num
1600 4 g/l ber of chlorophyll molecules per PSU) or a higher number
6 g/l of PSU within the cell, or both [10] , it always involves the
Irradiance (µmol photons/m 2 /s)
1400 10 g/l increase in cellular pigment (chlorophyll) content. During

photoacclimation, all the parameters that characterize the
1200 PI-curve change. When irradiance decreases, the quan
tum efficiency (a) increases, but Ik and Pmax decrease
1000 (Figure 6) , leading to a decreased capacity to use high
irradiances efficiently. These shade-adapted algae, when
800 (cyclically) brought to the surface layers, absorb photons
in large excess with three main negative consequences:
600
They waste most of the absorbed light energy because
they are photosaturated;
400
They may suffer from photoinhibition;
200
They strongly shade the cells in the layers below.
0
This is the reason why, even in optimally operated
0 2 4 6 8 10 12
algal cultures, large increases in irradiance lead to small
Culture depth (mm)
increases in productivity. Technology has not yet pro
vided a simple solution to this limitation. If, for exam
Figure 4. Light extinction with depth in Arthrospira
ple, high-light adapted algae (i.e., cells grown at high
platensis cultures at different biomass concentrations.
irradiance and characterized by small antennae) were
Culture depth is 1.2 cm and irradiance at the surface
used to inoculate a dense culture, they would readapt
1925 µmol m-2 s-1. The downwelling photosynthetic
to low light in a few hours [13] .
photon flux was measured with an underwater cosine
It is photosaturation or the LSE [11] that deter
corrected L1‑192 SA quantum sensor (Li-cor, Inc.,
mines the maximum efficiency that may be achieved
Lincoln, USA).
by an outdoor dense algal culture. The LSE may be
Data from [Chini Zittelli G, Tredici M, Unpublished Data].
quantified by the ‘Bush equation’:

Equation 1
f = 8` ln j + 1B
Is I0 Box 2. Light zones in dense algal cultures.
I0 Is In well-mixed mass cultures, microalgae are exposed to a complex, highly fluctuating light
field. Under conditions of high cell density, due to the self-shading of cells, light is attenuated
which allows calculation of the
exponentially through the culture according to the Lambert–Beer law (Figure 4), from full sunlight
‘light utilization efficiency’ ( f) or at the surface to darkness at the bottom. Although the decrease of irradiance is continuous along
the fraction of the MPE that may the culture depth, four main zones can be conceived in a dense algal culture exposed to strong
be attained based on the consid solar light (Figure 5):
eration that light energy is dis The surface layers in which irradiance (I) is above Ih and photoinhibition occurs;
sipated in the culture according The light saturated zone, in which irradiance is below Ih but above Is and the maximum
to Lambert–Beer’s law and that photosynthetic rate (Pmax) is achieved;
irradiances higher than Is are not The light limited zone, in which irradiance is between Is and Ic and light is used with
utilized (Figure 7) [11] . From the maximum efficiency;
Bush equation, we can derive that The dark zone in which irradiance is below Ic and net photosynthesis cannot take place.
The three first zones, in which light is sufficient for net photosynthesis, make up the photic
at low irradiances, f approximates
volume. It is noteworthy that the above description of light penetration in an algal culture is
1 and the MPE may be potentially
an oversimplification of the reality and we do not know what light climate algae really perceive
attained, but at high Io /Is ratios, f in the culture. Besides, the light penetration depth varies with wavelength: green light, poorly
falls rapidly (Figure 7) . At a value ofabsorbed by algal chlorophylls, penetrates 20-times deeper than blue or red light. For this
Io /Is of 20 (e.g., a typical microalga reason, it has been suggested that green light may have an important role in light limited dense
that saturates at ~5% full sunlight, algal cultures [26].
grown in an outdoor culture), f is
approximately 0.2, in other words, only 20% of the (diurnal photoinhibition) and measured a reduction
thermodynamic efficiency can be attained. In simple in daily productivity of approximately 20% [27] . Here
words, at Io values much higher than Is, photosatura a minimum loss of 10% is considered.
tion results in large losses of absorbed photon energy. The losses due to photosaturation are difficult to
Together with irradiance, the value of Is is thus a major quantify. Here, I will assume losses of 30% of PE,
factor in determining productivity of outdoor algal although much higher values have been suggested.
cultures and it appears clear from the Bush equa Overall, photosaturation and photoinhibition account
tion that there would be great advantage in growing for a minimum 40% decrease of PE in outdoor cul
algal species with high Is values [11] . With the exclu tures on sunny days. Thus, under the best conditions
sion of dinoflagellates, most marine algae saturate at considering reflection, respiration, photosaturation
irradiances below 100 µmol m-2 s-1 (~5% full sunlight). and photoi nhibition, but with no losses for photo
It is also necessary to consider that an outdoor cul respiration, we can expect a maximum PE of 5.4%
ture is subjected to varying irradiances at its surface: (12.4 × 0.9 × 0.8 × 0.60) on total impinging solar
from darkness to full sunlight (up to more than 2000 radiation by algal cultures (Table 1) .
µmol photons m-2 s-1 in clear summer days) to dark Overall, when all the possible losses are considered,
ness again in a few hours. This should alleviate photo it results that algae, even if not limited by photo
saturation and have a positive influence on PE, since respiration and without the burden of concentrating
higher efficiencies can be attained under the low irra CO2 mechanisms, are not superior to C4 plants in
diances of the early morning and late evening hour, terms of PE (Table 1) . This is reflected in the maxi
although there is evidence that outdoor cultures are mum productivities actually achieved by outdoor algal
strongly limited by their inability to quickly respond cultures (see following sections).
to diurnal irradiance changes [26] .
Full sunlight
Losses by photosaturation & photoinhibition in
I > Ih
algal cultures
Under full sunlight, the cells in the upper layers of
Is < I < Ih
an algal culture will be exposed to excessive irradi Growth zone
Ic < I < Is Mixing
ance and photoinhibited. Photoinhibition is common
to most algae at irradiances as low as approximately I < Ic
10% full sunlight [11] . Photoinhibited algal cells show
Algal cell
a much reduced photos ynthetic capacity both at low
and high irradiances (Figure 6) . Vonshak and Guy Figure 5. The different zones in a dense algal culture
were the first to demonstrate that dense A. platen- under full sunlight. Symbol explanation in Box 2.
sis cultures suffered from photoinhibition at midday

Review Tredici
cycle) [9,10,13] . On longer timescales, photosynthesis is

regulated through changes in cellular pigmentation
High-light acclimated cells and in size and number of the photosynthetic units and
Photosynthesis rate
their turnover rate. To what extent these mechanisms

Low-light acclimated cells operate in outdoor mixed algal cultures is difficult to
say. Experiments carried out with Dunaliella tertiolecta
under a fluctuating light regime that simulated mixed
Photoinhibited cells layers of turbid estuarine waters indicate that acclima
tion under fluctuating irradiance proceeds through
photoacclimation to accommodate to the longer periods
at low light and through rapid changes of nonphoto
chemical quenching to accommodate to high irradiance
Irradiance spikes [28] . This suggests a predominant influence of low
irradiance over high-irradiance stress in the adjustments
Figure 6. Changes in the light-response curve of photosynthesis as a to fluctuating irradiance. More in detail, the experi
consequence of photoinhibition and photoacclimation to low light in ments of Havelkova-Dousova et al. showed that, with
dense algal cultures. respect to cellular pigmentation, the cells acclimatized
to irradiances approximately three-times lower than
Algal cultures & photosynthesis under the average irradiance of the fluctuating regime [28] .
fluctuating light We can assume that, if the same mechanisms are at
The studies performed on photosynthesis under fluctu work in algae mass cultures exposed to high light, they
ating irradiance may greatly help our understanding of would seriously compromise the photosynthetic perfor
photoacclimation and regulation of photosynthesis in mance of the culture. Owing to the increased amount of
outdoor algal cultures. Phytoplankton in nature accli light-harvesting pigments, photon absorption is much
mate to irradiance changes over timescales of minutes increased in low-light-acclimated cells, but these cells
to hours. On shorter timescales they regulate photo have a reduced photosynthetic capacity (Figure 6) and
synthesis mainly by redistributing harvested energy this exacerbates photosaturation. Besides, low-light
between photosystems (state transitions) or by dissipa acclimated cells are more sensitive to photoinhibition,
tive nonphotochemical processes (e.g., the xanthophyll which leads to further reduction of the photosynthetic
performances (Figure 6) . The issue, however, remains
1.00 controversial and we cannot exclude that different
microalgae may adopt different acclimation strategies
(e.g., Grobbelaar et al. observed that in modulated light,
low-light-acclimated algae had higher photosynthetic
0.75
rates), which partially could explain the differences in
productivities attained with different species [29] .
Light utilization efficiency
The experiments of Havelkova-Dousova et al. sug

gest that algal cells exposed to the fluctuating light
regime of an outdoor culture would acclimate to low
0.50 light, since they reside for longer periods in the poorly
lit layers that prevail in the culture, while the very short
exposure to high light in superficial layers does not
promote long-term acclimation [28] . And this might be
0.25 the greater weakness of outdoor algae mass cultures: in
dense cultures, the bulk of photons that penetrate the
surface can be found ‘traveling’ in the top layers. This
means that most of the energy necessary for growth is
available in the region where it is largely in excess. Here,
0.00 photons are harvested by low-light-adapted cells under
0 10 20 30 40 50
I 0 /I S conditions of photosaturation, leading to low efficien
cies of conversion and, eventually, to photodamage.
Only a small fraction of (prevalently green) photons
Figure 7. Light utilization efficiency (f) in dense algal cultures quantified
reach the intermediate light-limited layers, where they
by the ‘Bush equation’ f = Is/Io [ (ln Io/Is) + 1].
are used with higher efficiencies. Then follows the dark

zone (at irradiances below compensation) allowing no C3 and C4 crops. In California (USA), productivi
net photosynthesis. The comparison with the light cli ties higher than 50 g m-2 day-1 have been reported
mate found in plant canopies as consequence of light with Sudan grass and maize [30] . Pennisetum typhoides
reflection and diffusion by leaves shows the superior (pearl millet) in West Australia has attained a yield
strategy of plants. It is reasonable to assume that the of 54 g m-2 day-1 and an efficiency of 4.5%. In the
culture density (combination of cell concentration and tropics, PEs of 4% have been achieved with napier
culture depth) giving the maximum productivity is the grass (El Salvador) and bullrush millet (Australia)
one that permits reaching the irradiance of compensa [30] . A 3-week mean productivity of 63.8 g m-2 day-1,
tion at the bottom of the culture. According to Raven, with a PE of 7.5%, was reported for sunflower [30] . A
to achieve maximum productivity it is necessary to hydroponic stand of Phragmites communis in Central
absorb no less than 95% of the impinging photons Europe attained an average of 57 g m-2 day-1 over a
[15] . Higher culture densities would increase the dark 130‑day growth season. Productivities above 60 g
zone, leading to higher levels of chlorophyll per cell and m-2 day-1 have been measured with the C3 Helianthus
lower efficiencies, besides increasing energy losses for annuus, Phragmites and Triticum and with several C4
maintenance. Outdoors, the situation is complicated grasses [31] . As seen at the short-term level and under
by the continuous variation of irradiance during the optimum growth conditions, there is no substantial
daylight period, and there have been no clear attempts difference between C3 and C4 plants. It is mostly
to continuously adjust the culture density to the chang the longer growing season duration and suitability
ing irradiance in order to establish the optimal density of local climate that determines the higher annual
at any time of the daylight period. In reality, even in yield of C4 plants. The aforementioned short-term
the laboratory under constant illumination it has been yields (>50 g m-2 day-1) if sustained, would lead to
observed that there is a range of cell concentrations, more than 180 tons of dry matter ha-1 year-1. Real long-
rather than a single specific value, leading to maxi term crop productivities never approach this limit.
mum productivities [25] . This said, in general, diluted Sugar cane, one of the most efficient crops known, has
culture perform better as far as they are able to cap been reported in few occasions to surpass 100 tons of
ture all the impinging photons. Both in the laboratory dry matter ha-1 year-1 with conversion efficiencies of
and outdoors, the highest productivities are commonly approximately 3.3% [32] , but typically produces less
achieved at dilution rates that allow growth rates of than 80 tons dry matter ha-1 year-1.
approximately half the maximum attainable [11,15,26] .
Productivity of microalgal cultures
Short- & long-term crop productivities A PE of 5% (a value that must be significantly reduced
Actual plant efficiencies in the field are much lower than if biomass instead of carbohydrate is considered)
those reported earlier for a number of reasons: would lead, on a clear summer day (> 24 MJ m-2), to
The growing season is reduced; storage of 1200 kJ m-2 of algae biomass and, with an
energy content of 20–23 kJ g-1 of dry biomass, to a
Only during a fraction of the growing season is the yield of 50–60 g m-2 day-1. There are many areas on
canopy cover ‘closed’; the earth that receive an annual average of more than
CO2 (at 380 ppm in air) is limiting (a still day can 22 MJ m-2 day-1 (~ 80,000 GJ per hectare and year) and
result in a considerable decrease in the CO2 thus have an annual productivity potential of 200 tons
concentration in the lower layers of a plant canopy); per hectare (Figure 8) . Unfortunately, long-term efficien
cies of 4–5% are never reached with algal cultures under
Plant growth is limited by various nutritional factors natural conditions.
and stresses (e.g., water shortage, far from optimal Potential productivities at two saturation irradi
temperature and pathogens). ances of 50 and 150 kJ m-2 h-1 (~65 and 190 µmol
High PE are commonly achieved in short-term photons m-2 s-1) were calculated by Rhyther consider
experiments under low irradiance. For example, one of ing the light transmission efficiency at the air–liquid
the highest PE (4.3%) was reported for sugar beet in culture interface, the maximum efficiency of conver
the UK. High productivities are typically achieved in sion of PAR in carbohydrate and the light utilization
the tropics using C4 plants, which are less likely to be efficiency in dense algal cultures (Bush efficiency fac
saturated under high light levels and temperatures, but tor) [11] . Without considering respiration and photo
efficiencies are not much higher. inhibition, maximum productivities of 30 and 60 g
In short-term experiments, where all the factors m-2 day-1 were predicted at the lower and higher satura
have been optimized, crop productivities above 50 g tion irradiance, respectively. The main conclusions of
m-2 day-1 and PE of 4% have been achieved both with the analysis were:

Review Tredici
The light saturation factor causes a flattening of the mixing [34] . The best published results in short-term
curves that relate productivity and irradiance, so that experiments and small ponds exceed 50 g m-2 day-1. In
any increase of daily irradiance does not lead to a Peru, with Scenedesmus obliquus, Heussler et al. attained
proportional increase in productivity (i.e., the rela a productivity of 53 g m-2 day-1 as an average of some
tionship between areal productivity and solar weeks [35] . In South Africa, productivities of up to 54
radiation is not linear); g m-2 day-1 and efficiencies of 4% have been measured
with the green microalgae Chlorella sp. and S. obliquus
The influence of Is increases with the increase of irra
[36] . It is interesting to note that, with few exceptions,
diance and at daily irradiances typical of high lati
the measured productivities of microalgal cultures are
tudes in summer, species with the lower Is would
not higher than the short-term yields reported for C3
achieve a 50% lower productivity. As mentioned: in
and C4 plants. This, in part, may be explained by the
outdoor cultures, relatively high photosynthetic effi
higher energy content typical of microalgae biomass
ciencies can be achieved only at low daily irradiances
(20–23 vs 18.5 kJ g-1 for plants). In commercial race
or by growing algae with high Is values. In any case,
ways ponds, yearly productivities have stalled below
the higher the daily radiation available, the lower the
10 g m-2 day-1, but the species commercially grown
efficiency of conversion. These predicted yields do not
(Dunaliella, Haematococcus and Arthrospira) are not
differ from the highest productivity values actually
particularly high producers.
measured in algal cultures.
Only few data are available for marine algae or
In outdoor algal cultures it is common to reach pro algae grown in seawater in long-term experiments.
ductivities of 15–25 g m-2 day-1 for 2–3-month peri Tredici et al. cultivated A. maxima (a nonmarine cyano
ods and of 30–40 g m-2 day-1 for shorter periods. With bacterium) in seawater plus urea as nitrogen source for
Tetraselmis suecica in carefully arranged annular-col 1 year, attaining an average yield of 7.4 g m-2 day-1 [37] .
umn setups, an average productivity of 38 g m-2 day-1 Materassi et al. attained a year-round productivity of
was measured in a 2-week period [33] . An average pro 14.2 g m-2 day-1 with Tetraselmis tetrathele grown in
ductivity of over 40 g (ash free dry wt) m-2 day-1 was board-mixed 160-m 2 ponds in Southern Italy [38] . It
achieved over a period of 1 month with T. suecica grown is estimated that under favorable climates and with
in a shallow flume containing foils to increase vertical suitable strains, productivities approaching annual
80
120
200
160 200
240
200
200 200
160
120
80
Figure 8. World map of algae biomass productivity (tonnes/ha/year) at 5% photosynthetic efficiency considering an energy
content of 20 MJ/kg dry biomass.

averages of 20–22 g m-2 day-1 (70–80 tons ha-1 year-1) by industry as research tools for Key term
are within our reach, but achieving them at a large scale biofuel production. PBRs have Raceway ponds: Widely used open
will be a major challenge and will require that all the recently been reviewed by Lehr and systems mixed by a paddle-wheel for
factors affecting growth (e.g., temperature, population Posten [44] , Carvalho et al. [45] and algae cultivation and commercial
production
density, mixing, nutrient supply and predator control) Tredici et al. [46] .
are optimized. Surpassing yields of 80 tons ha-1 year-1 Photobioreactors have many
will likely require genetically improved strains or other advantages over open ponds:
technologies able to counteract photosaturation and They are closed (but generally nonhermetically closed
photoinhibition (see the following sections). and nonaxenic), and thus limit direct exchange of
gases, liquids (e.g., rain) and particles (e.g., microbes,
Growth rate & productivity: an abused mismatch insects and dust) between the culture and the atmos
It is frequently reported that microalgae can grow 100- phere, making it easier to maintain a unialgal culture;
times faster than plants and double their mass in less
than 1 day and for this they would be the best candi Evaporation in PBRs is much reduced;
dates for biological production of fuels [39] . It is true that Due to a higher surface-to-volume ratio, PBRs may
some algae display high growth rates. Actually, some attain higher volumetric productivities than ponds [6]
of them can double their cell number (and biomass) in and adopt higher cell concentrations, which reduce
3.5 h. But what is often ignored is that high growth rates the costs for medium preparation and handling and
are not necessarily associated with high productivity, that for harvesting;
which is what really matters. The specific growth rate
(µ) measures the increase of biomass per unit time and They provide a more accurate control of culture
the unit biomass, and is one of the two variables that parameters;
determine productivity (i.e., the amount of biomass pro Elevated PBRs can be oriented and tilted at different
duced per unit volume or unit land area, per unit time). angles so as to maximize light capture and productiv
Productivity (Y) is calculated by multiplying the growth ity per square meter of reactor’s illuminated surface
rate by the actual amount of biomass (X) that is increas or, otherwise, so as to dilute light (i.e., reduce the
ing at that rate (Y = µX). To reach maximum growth irradiance at the reactor’s surface) and maximize PE
rates, algae need to be photosaturated. However, mass (see section: ‘Solutions proposed to overcome the LSE
cultures are typically photolimited because to maximize and photoinhibition in algae mass cultures’).
areal productivity there is the need to absorb all the
impinging photons. Thus, due to photolimitation, the The ultimate and most important advantage of PBRs,
cell growth rate in algae mass cultures is much slower however, is that they permit cultivation of algal species
than the maximum (typically approximately half the that cannot be grown in open ponds.
maximum). We could keep a culture in a pond at its Vertical reactors intercept sun rays at large angles and
maximum growth rate, but the cell density would be dilute light compared with horizontal ponds. Besides,
so low that most of the impinging photons would pass their rear surface receives mostly diffuse and reflected
through unused [202] . To increase productivity, we need radiation of low intensity. For this reason, vertical PBRs
to increase the cell concentration up to the point where are more efficient than horizontal ponds in terms of
all the impinging photons are captured. This increases solar-energy utilization, but it is not easy to devise strat
self-shading and decreases growth rate. Overall, the egies to deploy them so that this advantage is translated
higher the cell density, the lower the growth rate. into a practical benefit: higher productivity per unit of
occupied land surface area. The closer they are set up,
Photobioreactors & ponds the higher the productivity per land area occupied will
Several types of photobioreactors (PBRs) have been be, but also the higher the cost of the installation [6] .
designed and experimented with since the late 1940s, PBRs may be more productive than ponds in terms of
when algal cultures were first considered the ideal solar occupied land area, but they are not necessarily so. It
technology to produce biomass and protein on a large greatly depends on the strain requirements for light and
scale in a cost-effective manner and save the hungry temperature, on the reactor design and on season and
part of humanity [40] . PBR design and engineering is location [33] .
still a very active field of research, since closed culture Photobioreactors pay heavily for their advantages.
systems are necessary to grow photosynthetic microbes In general, PBRs are much more expensive to build
and exploit them as a source of aquaculture feeds, food than ponds, but simple low-cost systems can also be
additives, specialty chemicals, cosmetics and bioac designed. Tredici et al. have recently patented a panel
tive and labeled molecules [41–43] , and are preferred reactor made of a disposable polyethylene film that costs

Review Tredici
approximately €5 per square meter [101] . A well-designed was able to respond to nutrient stress with a significant
raceway pond made with a durable liner will not cost less increase of lipid synthesis. In a 20-l flat panel under
[203] . Although construction costs of PBRs may be simi artificial illumination, its fatty acid content increased
lar to those of ponds, their operating costs are not [44] . to more than 50% (on dry weight) during nitrogen or
Suitable mixing to provide the necessary mass transfer phosphorus starvation [3] . Outdoors, in a 110-l GWP
in air-bubbled green-wall panels (GWPs) and annular under nitrogen starvation, lipid productivity doubled
columns requires more than 100 W m-3 (approximately (from ~100 to 204 mg l-1 day-1) and the lipid content
2000 MJ ha-1 day-1) or up to 50% of the energy stored reached 60% of the dry biomass. In a two-phase cul
in the biomass [Bassi et al., Unpublished Data] . Mixing in tivation process (a nutrient-sufficient phase to produce
tubular reactors to achieve sufficient fluid velocities and the inoculum followed by a nitrogen-deprived phase to
Reynold’s numbers may require even higher energy boost lipid synthesis), a lipid production potential of
inputs [42,44] . Besides, there is the need for cooling, more than 90 kg ha-1 day-1 was measured. The experi
which is generally provided by water (even seawater) ments also showed that lipid accumulation was due to
spraying or by insertion of a cooling serpentine in the de novo synthesis and suggested that a sudden increase
culture [3] , again with not trivial energy expenditures. of light per cell, provided through culture dilution, was
Growing algae in open ponds is, in general, much necessary. The accumulated lipids in the nutrient-starved
cheaper. Mixing in raceway ponds may be achieved with biomass were mostly nonpolar triacylglycerols (TAGs).
an energy input of approximately 200 MJ ha-1 day-1 [47] , According to these results, Nannochloropsis sp. F&M-
which represents an expenditure of 3–5% of the energy M24 has the potential for an annual oil production of
fixed in biomass and there is no need for cooling, which 20 tons per hectare in the Mediterranean climate. This
(in dry climates) is provided by natural evaporation. is four-times the productivity achievable by oil-palm in
For these two reasons, the author does not see at pres the tropics. The process described earlier is well suited
ent the possibility that PBRs may substitute raceway to be performed in two phases using both PBRs and
ponds in the bulk production of biomass. Nevertheless, ponds. The first nutrient-sufficient phase for biomass
some recent analyses seem to support the conclusion production is to be carried out in PBRs, where condi
that algae biomass production at a large scale will be less tions are more carefully controlled and the culture health
expensive using PBRs than open ponds [48] . and proper biomass composition can be ensured, while
Ponds are cheaper to operate than PBRs, but suffer limiting the risks of contamination. The second phase for
from various limitations [203] . The main disadvantage oil accumulation, where nutrients are not provided since
is that, being open to the atmosphere, ponds are much growth is not necessary, is realized in open ponds. The
more susceptible to contamination. Unwanted algal oil accumulation phase takes 3–4 days, so risks that the
species, fungi and grazers will inevitably enter into culture is compromised by contaminants or grazers are
the culture and compete with the inoculated selected nil or much reduced.
strain [2,6,26] . Furthermore, one of the advantages of There are many other marine algae that, similar to
ponds, evaporative cooling, may become a drawback Nannochloropsis F&M-M24, show a high potential for
where freshwater is scarce [2] or when seawater cannot be oil production [1–3] . They are productive and their lipid
used to replace evaporation because the growth medium content can be modulated by varying nutrient supply or
must be recycled. Very likely, an industrial plant for bio by means of stresses. Some algae have shown lipid con
fuel or protein production from microalgae will require tents of up to 85% of dry weight, with the bulk of accu
a combination of PBRs (for inocula production) and mulated lipid as nonpolar TAGs, the best substrate for
ponds (for bulk cultivation). biodiesel. It is highly likely that among the several thou
sand species and billions of strains that colonize almost
Microalgae for oil every niche of the earth, many organisms suitable to
Biofuels from microalgae seem to be the only renew outdoor mass culture and biofuel production might be
able fuels with the potential to displace fossil oil-derived found. It is worth mentioning that the selected strain,
transport fuels without jeopardizing food supply [49] . besides being productive and with a constitutional high
Recently, 30 microalgal strains were screened in the lipid content or able to respond with oil accumulation
laboratory for their biomass productivity and lipid con to stresses, must be robust enough to withstand mixing-
tent in order to evaluate their oil production potential [3] . generated shear-stress and to adapt to the rapid changes
As expected, lipid-rich strains showed, in general, lower in physicochemical parameters of outdoor ponds. Other
biomass productivity. Among the strains that showed a nonsecondary traits are easy harvestability and extract
good combination of the two parameters and attained ability, characteristics not present in Nannochloropsis sp.
the highest lipid productivity, the eustigmatophyte F&M-M24, which is only 3–5 µm in size and endowed
Nannochloropsis sp. F&M-M24 was chosen because it with a hard cell wall.

Another interesting marine candidate for oil pro past a plant made of 12 m high, 0.9 m diameter vertical
duction is Tetraselmis, even if the basal lipid content columns, currently AlgaeLink proposes large tubular
of many members of this genus is not high (8–20% of serpentine reactors. The expected volumetric and areal
dry weight) [3] and strains typically do not accumulate productivities exceed the theoretical photosynthetic
oil during stress, because they preferentially respond efficiency [46] . It seems that at the basis of the wrong
with carbohydrate synthesis. With some good strains projection was the assumption that, since algae in the
of Tetraselmis that show high biomass productivity and laboratory double in 24 h they will do so also outdoors
stability in open ponds [34,38] , it is reasonable to expect and at any cell concentration or standing crop, which
a yield of more than 70 tons of dry biomass ha-1 year-1, is not true.
of which 15 tons may be oil and 30 tons protein. Many other companies have been formed in recent
However, despite the extensive efforts underway, the years to exploit the growing interest in microalgae origi
potential of algae to achieve commercial-scale produc nated by increasing oil prices and policies in support
tion of oil remains doubtful. The main reason for this is of biofuels. Not all of them claim unrealistic yields,
that no large-scale production facilities are in operation but, with few exceptions, they have not carried out the
yet. Consequently, no data on large-scale productivi extended large-scale tests necessary to prove the pro
ties and costs of production, as well as operation of the posed technology. So far most of the PBRs are in a proto
facilities, exist. type stage and productivities for a commercial-scale PBR
are still unknown. In reality, no company has, at present,
Photobioreactors for biofuels a mature technology to be on the market and compete
In the last years, numerous startup firms in the micro with fossil fuels. However, some of the proposed designs
algae biofuel field have proposed new ideas and inno are worth being improved [44] and it is not to exclude
vative applications of old PBR designs. Among them that after adequate R&D these or other systems, might
GreenFuel Technologies Corp. (GFT; MA, USA) was be industrialized and used, likely in combination with
founded in 2002 by MIT and Harvard scientists. With open ponds, to produce algae biofuels.
its triangular airlift reactor, GFT claimed to be able to
capture more than 80% of the CO2 and NOx from coal- Species selection & control
fired station gases and attain biodiesel productivities The issue of strain selection deserves special attention,
of 80 tons ha-1 year-1 [46] . This system was not further since productivity and the capacity to maintain uni
developed and a different design called the 3D-Matrix algal cultures largely depend on the chosen organism.
System was tested at GFT facilities in the Arizona des The algal strain to be cultivated should be selected on
ert [204] . With the 3D-matrix in the summer of 2007, the basis of many other criteria besides oil content and
according to company press releases, an average areal productivity, among which there are resistance to con
productivity of 98 g (ash free dry wt) m-2 day-1 was tamination, tolerance to high oxygen levels and tempera
achieved, with peak productivities of over 170 g (ash free ture variations and harvestability [203] . For large-scale
dry wt) m-2 day-1 on good sunny days. The peak pro biofuel production a series of PBRs of increasing size and
ductivity corresponds to an efficiency of approximately decreasing level of containment and cost will be neces
18% of total solar radiation, which exceeds the MPE sary to provide the large amount of inocula required
calculated for biomass production by 80%. for the ponds [2,203] . Starting the process with axenic
Valcent Products, Inc. (Vancouver, Canada) has (pure) culture is of dubious efficacy. Axenic cultures
developed a vertical bioreactor (the ‘vertigro’) con are unstable and sooner or later, these cultures will be
sisting of a series of closely spaced vertical bioreactors contaminated by airborne bacteria and other organisms,
made from thin plastic-film membranes. The culture is which are not necessarily beneficial. On the contrary, it
pumped to the top of the reactor and circulates by grav might be an advantage if with the inoculum the algae
ity to the bottom in a meandering way. The technology carry their ‘own’ microflora. The relationships between
is expected to yield 150,000 gallons of oil acre-1 year-1 algae and bacteria are complex and not well understood,
(about 1,400,000 liters ha-1 year-1), which is thermo but evidence is growing that ‘beneficial’ associated bac
dynamically not possible. The system is interesting for teria are necessary to maintain a healthy algae popula
its simplicity and verticality. Problems might arise at tion, even more so in the harsh environment of an open
high cell densities that favor biofouling and because of pond [43] .
oxygen build-up [46] . There exist strong motivations for maintaining the
AlgaeLink N.V., based in The Netherlands, commer selected algal species in the culture. Biomass separa
cializes PBRs for biodiesel production from microalgae tion and processing require a species-specific approach.
from demonstration size up to systems that are claimed Harvesting techniques and their efficiency vary
to produce 100 tons day-1. After having marketed in the with the size, shape and buoyancy of the cells. Some

Review Tredici
species are easier to flocculate or sediment than others. light, each photosynthetic unit captures only one photon
Sedimentation velocity is proportional to the square of and the slow electron transfer between the photosystems
the radius of the cell, thus larger cells should be selected and the dark reactions leading to CO2 fixation can be
to favor sedimentation. Cell wall thickness and robust completed in the subsequent dark phase. Unfortunately,
ness influence extraction and vary from species which conventional mixing can do little to improve the effi
have no actual cell wall (e.g., Dunaliella) to species such ciency of conversion of light through the ‘flashing light
as Chlorella and Nannochloropsis, which have a conspicu effect’. Achieving a light–dark cycle overlapping with the
ous hard cell wall. Selected strains must be maintained timescale of the flashing light would require very high
in culture because optimization of cultivation conditions (ordered) mixing, which is technically not feasible in
is strain-specific. Every strain has its optimum tempera algal cultures [13,51] . Medium and low frequency light-
ture, pH, salinity, nutrient concentration and specific dark cycles of 100 ms to 1 s have been reported to lead to
capability to resist hydrodynamic stresses, tolerate high high efficiencies [34] , but the results are inconclusive and
oxygen concentrations and compete with contaminants. it cannot be excluded that the beneficial effects observed
If the biomass is intended for human use, the strain have to be ascribed to other factors, such as preventing
would have to be subject to extended toxicological tests settling or improved mass transfer [51–53] . Under short
and of course the strain cannot be changed during culti light–dark cycles of 94/94 ms and at an irradiance of
vation. The same is true if the strain was chosen because 445 µmol m-2 s-1, Jansen et al. observed with D. tertiolecta
its oil composition complies with standards. The dif a significant increase of PE compared with continuous
ficulty of maintaining selected algae for long periods in illumination [54] . However, the PE did not increase under
open ponds has been somewhat overestimated, perhaps light–dark cycles of 31/156 ms at 1025 µmol m-2 s-1. The
as a result of the Aquatic Species Program of the US authors suggest that at high irradiances, the length of
DOE carried out from 1978–1996 [205] , in which hun the light period must be less than 10 ms to obtain a
dreds of algal strains were tested and none has shown benefit in terms of PE. However, this is difficult and
the capacity to stay in culture for long. The difficulty very expensive to attain in algae mass culture.
of maintaining an unialgal culture is much related to
seasonal variability. In Southern Italy, cultures of the Light dilution
marine T. tetrathele and of the nonmarine A. platensis, The problem of the LSE in algae mass cultures was rec
have been kept in open ponds in seawater for more than ognized very early and light dilution, in time and in
1 year [37,38] , without experiencing crashes or devastating space, was among the first solutions to be proposed [40] .
contaminations. However, under prolonged unstable or Spatial light dilution means distributing the impinging
rainy weather conditions, maintaining unialgal cultures photon flux on a larger surface area so as to decrease irra
at the large scale may represent an impossible task, even diance and increase PE. Following the first approaches
with the help of regular use of massive inocula. But it that led to not conclusive results, this field has remained
would be unwise to select such climates for large-scale quiet for some decades [23] . Researches started again
algae mass cultures in ponds. in the early 1980s essentially with the use of elevated
reactors (panels, coils and biofences). Almost all of the
Solutions proposed to overcome the LSE & experiments showed substantial increases of PE and pro
photoinhibition in algae mass cultures ductivity with light dilution [23,33,51,55] . The main draw
Various solutions have been proposed to overcome back of this approach is that to dilute light significantly
the deleterious effects of photosaturation and photo with respect to that impinging on the horizontal, similar
inhibition and to increase productivity of outdoor algal to plants that adopt high leaf areas per unit land area, the
cultures. Strong mixing, generation of algal mutants ratio between the illuminated surface area of the system
with reduced pigment content that do not acclimate and the occupied land surface area must be substantially
and light dilution will be briefly described in the greater than one and this requires a greatly increased
following sections. surface area of reactor’s transparent walls and supporting
structures, which enhance capital costs. The concept of
Rapid mixing light dilution evolved and has led to fiberoptical-based
Rapid mixing to increase PE was proposed very early systems, in which light capture and use are physically
since it was expected to create the ‘flashing light effect’ separated. Visible solar light, collected by mirrors, is con
first demonstrated by Kok [50] . Kok observed that when centrated through lenses and delivered into the bioreac
high-intensity light is provided in flashes of milli tor via an array of flexible optical fibers, which transmit
second duration, followed by a five to tenfold longer photons via total internal reflection. Because the solar
dark period, a high efficiency of energy conversion can collectors track the sun, the main significant variation
be achieved. The reason is that during a brief flash of in photon flux inside the reactor is cloud cover.

Wijffels and his team at the Bioprocess Engineering assembly of the photosystems in chloroplasts [58] . Even
Group of the Wageningen Universit y (The if this goal has not been achieved yet, it has been dem
Netherlands) conceived a rectangular airlift PBR that onstrated that mutants with reduced antenna size may
can possibly be scaled up to more than 100 m 3 of achieve a 50% higher productivity under high irradi
culture volume [51] . The system is composed of a large ance compared with the wild type [59] . Truncated Chl
number of light-redistributing plates fixed a few cen antenna-size mutants have been recently tested in mass
timetres from one another. The plates are connected cultures under sunlight in a greenhouse, confirming the
to the sides of the reactor and to optical fibers, which results obtained in the laboratory [58] . This technology
transmit light collected in the field by parabolic dishes. holds great promise, even if it still needs to be confirmed
Mixing, provided by air injection between adjacent in long-term experiments under full sunlight.
plates, can be adjusted so that culture hydrodynam
ics and the light–dark cycles to which the cells are Future perspective
exposed are similar to those found in flat panels. It is Phototrophs have evolved their mechanisms for solar
estimated that by collecting sunlight from a 2-ha field energy capture and storage under conditions in which
a daily average of 1200 µmol photons m-2 s-1 could flexibility to adjust to the changing environment was the
be supplied at the surface of the light-redistributing primary need, while the selective pressure to maximize
plates and a productivity of 200 tons (dry wt) year-1 the efficiency of solar energy conversion was minimal.
attained [51,56] . This high-technology design would The photochemistry of the initial light-to-charge con
be very expensive to build, but in time the cost of version reactions (not including losses in the antenna
lenses, mirrors, solar-tracking devices and optical and in CO2 fixation) in the photosynthetic membrane
fibers is expected to decrease thanks to mass produc shows a very high efficiency (~50% using red pho
tion. More recently, the same group developed the tons) [60] , but large inefficiencies emerge when this
Green Solar Collector, a PBR in which sunlight is energy is stored in the form of carbohydrate (see the
captured through polymethylmethacrylate (PMMA) paragraph on ‘legacy biochemistry’ in [61]). By mim
Fresnel lenses that are able to rotate over two axes to icking photosynthesis in artificial systems, the unde
follow the sun [57] . By rotating, the lenses also main sirable aspects of natural energy conversion might be
tain the line of focus on top of PMMA light-guides. overcome. Artificial photosynthesis provides oppor
Light refracts into the rectangular part of the light- tunities for human ingenuity to make a tremendous
guides and propagates by total internal reflection with advance in solar energy capture and conversion [61,62] .
negligible attenuation. Refraction out of the terminal However, while some progress has been made in the
narrowed part of the guide distributes photons into field, researchers have not yet developed the necessary
the culture compartment. Irradiance at the light-dis efficient and robust components for incorporation into a
tribution surface of the guide will be about half that working system [62] and for many years we will rely still
of sunlight, thus substantially increasing PE [57] . on the products of natural photosynthesis (both fossil
Cultivation of microalgae in optical-fiber PBRs and renewable) for our energy needs.
has been demonstrated only at the laboratory scale Microalgae are not more efficient solar energy con
and the very high cost of solar tracking devices and verters than plants. Biomass productivities of 80 tons
distribution systems seem prohibitive, but the poten ha-1 year-1, which are in the range of high yields attained
tial of this technology is high and worthy of further with C4 crops in the tropics, must be considered as the
R&D efforts. maximum achievable at large scale with microa lgae. It
is this number (or more prudently a lower one) that
Mutants must be used in determining the feasibility of any
In algae, up to 350 Chl a and b molecules can be found microalgae venture.
in association with PSII, while PSI may contain up to In recent years, unrealistic productivities exceeding
300 mainly Chl a molecules [58] . These large assemblies 200 tons ha-1 year-1 have often been claimed or thought
of light harvesting complexes give a competitive advan possible with microalgae. These claims have influenced
tage in nature, where light often limits growth, but they even the European Community that in the last call under
are the cause of photosaturation and photoinhibition FP7 Topic ENERGY.2010.3.4–1: ‘Biofuels from algae’
in mass cultures. A practical solution to the LSE would opens to proposals aiming at demonstration plants (mini
be the use of algal mutants with smaller amounts of mum: 10 ha) and productivities in the range of 90–120
antenna chlorophylls and accessory light-harvesting pig dry biomass tons ha-1 year-1. These expected productiv
ments. The objective of the researchers in this field is to ity values would, with current technologies, exclude the
minimize the antenna size to 37 Chl molecules for PSII whole of Europe as possible production sites and are hard
and 95 for PSI, the minimum size that would permit to attain even in the most insolated places of Africa.

Review Tredici
Today, there is a renewed interest in microalgae that after month, cost-effectively supplying CO2 and nutri
reminds of the surge of algae popularity in the 1950s ents, providing mixing, pumping water, harvesting and
inspired at that time by their high protein content. processing the biomass are the major issues on which
Consequently, large amounts of resources (financial our efforts must be focused. Fortunately, there exist
and human) are currently available for R&D in this no absolute impediments that suggest that either the
area. We, as algologists, have a unique opportunity to biological or engineering advances required to overcome
help redesign the goals of microalgal biotechnology: no these obstacles cannot be eventually achieved. It will
more exclusively centered on high-cost specialty chemi require considerable investment and R&D effort, but
cals as in the three past decades, but a novel ‘green’ in a decade, or even less, ‘mariculture on land’ may
technology able to contribute to develop a whole range become a reality.
of novel foodstuffs and renewable nonfood commodi
ties in a sustainable way. Besides biofuels, microalgae Acknowledgements
can provide proteins, carbohydrates and raw materi The author is grateful to Juergen Polle, Associate Professor in the
als for the manufacture of a spectrum of commodity Department of Biology of the Brooklyn College of the University of
chemicals (industrial starches, fatty acids, paper and New York, USA and to Graziella Chini Zittelli, researcher at the
plastics) today obtained from petrochemicals or plants. Consiglio Nazionale delle Ricerche of Italy, for critically reading the
The current price gap (indeed quite large) with petro manuscript and helpful discussion.
chemically derived products will gradually erode, as
petroleum reserves become depleted and microalgae Financial & competing interests disclosure
technology advances. The author is a consultant of companies in the field, a member of the
Mariculture on land may not save the planet, but scientific advisory board of Aurora Biofuels and member of the Board
might have a place in the ‘next green revolution’, because of Fotosintetica & Microbiologica. The author has no other relevant
it can contribute to cover part of our food and energy affiliations or financial involvement with any organization or entity
needs without further impoverishing fertile soils and with a financial interest in or financial conflict with the subject
freshwater resources and jeopardizing natural eco matter or materials discussed in the manuscript. This includes
systems. For this to be done, the numerous obstacles that employment, consultancies, honoraria, stock ownership or options,
hinder the technology must be tackled and solutions be expert testimony, grants or patents received or pending, or royalties.
provided. Maintaining selected strains in open ponds of No writing assistance was utilized in the production of
hectares, achieving sustained high productivities month this manuscript.
Executive summary
A minimum of eight photons is required to fix one molecule of CO2 into carbohydrate through oxygenic photosynthesis.
Quantum requirements lower than eight have been measured, but not substantiated.
The maximum (theoretical) photosynthetic efficiency of oxygenic photosynthesis is 12.4% (on total solar radiation).
Under full sunlight, photons are absorbed at a rate that largely exceeds the capacity of the photosynthetic machinery, leading to
photosaturation and photoinhibition.
Owing to photosaturation and photoinhibition and other unavoidable losses, the MPE of algal cultures under full sunlight is reduced to
approximately 5%.
The highest productivity values measured under natural conditions, both in short- and long-term experiments, with microalgae and plants
do not differ significantly.
Closed systems for algae cultivation (photobioreactors) require high energy inputs for mixing and cooling.
Large open ponds have a more favourable net energy ratio, but are very unstable ecosystems.
Microalgal cultures can attain much higher oil and protein yields than crop plants.
Selected strains and solutions to the light-saturation effect are necessary to produce cost-effective algae food, feed and biofuels .
Microalgae biofuels have high potential, but more R&D is necessary to overcome the obstacles that hamper the scaling-up of
microalgal cultures.
Despite their limitations, microalgae remain one of the most attractive sources of next-generation biofuels as they can be grown in saline
or seawater on nonarable lands.
land-based crops for biofuels. 3 Rodolfi L, Chini Zittelli G, Bassi N et al.

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Photobiology of microalgae mass cultures: understanding

Biofuels (2010) 1(1), 143–162

144 Biofuels (2010) 1(1) future science group

Pheo FeS A ATP-synthase

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region of the PI-curve.

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1400 10 g/l increase in cellular pigment (chlorophyll) content. During

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cycle) [9,10,13] . On longer timescales, photosynthesis is

their turnover rate. To what extent these mechanisms

The experiments of Havelkova-Dousova et al. sug­

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land-based crops for biofuels. 3 Rodolfi L, Chini Zittelli G, Bassi N et al.

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39 Murphy DJ. Biotechnology: its impact and n

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