Bioresource Technology 125 (2012) 75–81

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Spectral conversion of light for enhanced microalgae growth rates

and photosynthetic pigment production
Seyedeh Fatemeh Mohsenpour a, Bryce Richards b, Nik Willoughby a,⇑
a
Institute for Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS, UK
b
Institute of Photonics and Quantum Sciences, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS, UK

h i g h l i g h t s

" Growth/chlorophyll production of microalgae under varied light conditions studied.

" Natural light source modified using various luminescent acrylic sheets.
" Improved growth rates achieved under wavelength-modified light.
" Chlorophyll-a production increased under wavelength-modified light.
" Improved growth under modified natural light reduces need for artificial lighting.

a r t i c l e i n f o a b s t r a c t

Article history: The effect of light conditions on the growth of green algae Chlorella vulgaris and cyanobacteria Gloeothece
Received 15 June 2012 membranacea was investigated by filtering different wavelengths of visible light and comparing against a
Received in revised form 16 August 2012 model daylight source as a control. Luminescent acrylic sheets containing violet, green, orange or red
Accepted 19 August 2012
dyes illuminated by a solar simulator produced the desired wavelengths of light for this study. From
Available online 31 August 2012
the experimental results the highest specific growth rate for C. vulgaris was achieved using the orange
range whereas violet light promoted the growth of G. membranacea. Red light exhibited the least effi-
Keywords:
ciency in conversion of light energy into biomass in both strains of microalgae. Photosynthetic pigment
Microalgae
Light wavelength
formation was examined and maximum chlorophyll-a production in C. vulgaris was obtained by red light
Luminescent illumination. Green light yielded the best chlorophyll-a production in G. membranacea. The proposed illu-
Photosynthetic pigments mination strategy offers improved microalgae growth without resorting to artificial light sources, reduc-
Solar ing energy use and costs of cultivation.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction by microalgae cells thus the quality of light in terms of intensity

Cultivation of photosynthetic microalgae has received increased
attention as a potential source of high-value biochemical compo-
nents such as natural colourants, polyunsaturated fatty acids, pro-
teins and polysaccharides (Chen et al., 2010a,b) as well as being a
potential biofuel source or food material. In addition microalgae
cultivation has been considered as greenhouse gas mitigation
strategy in which solar-driven cells capture carbon dioxide (CO2)
and convert it into organic chemicals (Chisti, 2007).
In photoautotrophic cultivation mode light is the main source of
energy and inorganic carbon (such as CO2) is used as the carbon
source (Huang et al., 2010). Photons can be absorbed as nutrient
⇑ Corresponding author. Tel.: +44 (0) 131 451 4377; fax: +44 (0) 131 451 3129.
E-mail addresses: sm627@hw.ac.uk (S.F. Mohsenpour), B.S.Richards@hw.ac.uk
(B. Richards), N.A.Willoughby@hw.ac.uk (N. Willoughby).
and wavelength is critical for cell growth (Wang et al., 2007).
Specific growth rate and photosynthetic pigment formation are
highly influenced by the light source. Up to date the only light
sources which have been used for illumination of microalgae cul-
tures and capable of emitting light in specific wavelengths are light
emitting diodes (LEDs) (Wang et al., 2007). Long life expectancy,
low heat generation and efficient light conversion are the advanta-
ges of using a light source such as LED with selective wavelengths
(Chen et al., 2010a,b). However, to date no research has been fo-
cused on using luminescent acrylic sheets as a tool for selecting
certain wavelengths of sunlight for illumination of microalgae cul-
tures. The proposed technique uses transparent thermoplastic
polymethyl methacrylate (PMMA) doped with fluorophores
against a solar simulator to filter out the undesired wavelengths.
Fluorophores contain luminescent molecules and depending on
their colours they absorb light at specific wavelengths and re-emit

0960-8524/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.08.072
76 S.F. Mohsenpour et al. / Bioresource Technology 125 (2012) 75–81

them at longer wavelengths. PMMA is a common material for con- were designed to mimic local natural growth conditions, to repro-
struction of photobioreactors and hence is an ideal material for duce as best as possible a potential local commercial operation.
investigation of growth conditions related to light wavelength
variation.
2. Methods
Luminescent acrylic sheets have been used for building lumi-
nescent solar concentrators (LSC). LSCs are simple devices used
2.1. Microalgae strain
for absorbing incident solar radiation and emitting fluorescence
photons (Wilson et al., 2010) and were introduced more than three
C. vulgaris (CCAP 211/79) cultivated in bold basal medium with
decades ago for concentrating sunlight (Goetzberger and Greube,
3-fold nitrogen and vitamin (3N-BBM+V) was obtained from waste
1977; Ho et al., 2012). When the transparent sheet receives the
solvent biofilter at Heriot-Watt University, Edinburgh, UK. Medium
incident light on the front surface, the photons of sunlight are ab-
(3N-BBM+V) was autoclaved and contained the following compo-
sorbed by fluorescent dye molecules. The dye re-emits the cap-
nents: 25.0 g L1 NaNO3; 2.5 g L1 CaCl22H2O; 7.5 g L1 MgSO4-
tured photons and transports them towards the edges of the
7H2O; 7.5 g L1 K2HPO43H2O; 17.5 g L1 KH2PO4; 2.5 g L1 NaCl;
sheet via total internal reflection. The transmitted photons are then
trace element solution (FeCl36H2O, 97.0 mg L1; MnCl24H2O,
collected by solar cells which are attached to the edges of the LSC.
41.0 mg L1; ZnCl2, 5.0 mg L1; CoCl26H2O, 2.0 mg L1; Na2MoO4-
A LSC is a ‘‘non-imaging’’ concentrator meaning that it does not use
2H2O, 4.0 mg L1); Vitamin B1 (0.12 g Thiaminhydrochloride in
lenses, mirrors or combination of both to collect and concentrate
100 mL distilled water. Filter sterile), Vitamin B12 (0.1 g Cyanoco-
sunlight. Whilst collection of re-emitted photons around the edge
balamin in 100 mL distilled water, take 1 mL of this solution and
of an luminescent sheet is not a practical arrangement for con-
add 99 mL distilled water. Filter sterile).
struction of a large object such a photobioreactor, a proportion of
G. membranacea (CCAP 1430/3) cultivated in blue green algae
the re-emitted photons along with the vast majority of the trans-
medium (BG 11) in the bio-processing laboratory of chemical engi-
mitted photons pass directly out of the face of the luminescent
neering department at Heriot-Watt University, Edinburgh, UK.
sheet and these can be utilised for algal growth.
Medium BG 11 contained the following components: 15 g L1
There are different types of fluorophores which have been used
NaNO3; 4.0 g L1 K2HPO4; 7.5 g L1 MgSO47H2O; 3.6 g L1 CaCl2-
in luminescent sheets. Three main categories of fluorophores are
2H2O; 0.6 g L1 Citric acid; 6 g L1 Ammonium ferric citrate green;
organic dyes, rare earth materials and quantum dots (Rowan
0.1 g L1 EDTANa2; 2.0 g L1 Na2CO3; Trace metal solution (H3BO3
et al., 2008). Photo-stability is an essential factor for performance
2.86 g L1; MnCl24H2O 1.81 g L1; ZnSO47H2O 0.22 g L1; Na2-
of organic dyes. The main problem using original organic dyes
MoO42H2O 0.39 g L1; CuSO45H2O 0.08 g L1; Co(NO3)26H2O
was that they had poor photo-stability capable of performing for
0.05 g L1). Medium BG 11 was autoclaved and the pH was ad-
only a few weeks under solar irradiation (Hermann, 1982).
justed to 7.1.
Photo-stability of luminescent sheets with different host mate-
rials and fluorophores has been investigated in the literature (Her-
mann, 1982). New fluorophores with stability of several years 2.2. Light sources
under light exposure were later introduced and studied. For in-
stance Lumogen F range dyes (BASF) with great photo-stability 150 W Xenon arc lamps (Luxtel’s, Ceralux) were driven by a
and high quantum yields were developed and used for various so- 300 W power supply (PS300-12, Perkin–Elmer). Xenon arc lamps
lar applications including day-lighting system (Sousa et al., 2012). produce a broad illumination spectrum including short wave-
For this study, organic fluorescent dyes were selected as they show lengths of ultra violet (UV), visible (PAR) and long wavelengths
great solubility in different types of organic polymers such as of infra red (IR) and are commonly used as in solar simulators for
PMMA which is widely used in various photobioreactor designs. the photovoltaic industry and a typical spectrum of a solar simula-
As microalgae use sunlight only in the photosynthetic active tor can be found in (Vejrazka et al., 2011). The output spectrum of
radiation (PAR) range which includes wavelengths between 400 the lamps was measured using a high resolution spectrometer
and 700 nm, using an LSC as a new illumination strategy seems (HR2000 Ocean Optics, USA). The output spectrum of the lamps
an interesting research topic which is worthy of further investiga- using the described light filters is presented in Fig. 1.
tion. Previously a study focused on optimisation of light quality
and quantity for plant growth used fluorescent films with different
fluorescent pigments (Chrismadha and Borowitzka, 1994). The re-
sults of the study showed that blue fluorescent films enhanced the
growth of strawberry fruit whereas red fluorescent films delayed
the fruit production considerably. The study suggested that using
blue fluorescent films in greenhouses could potentially promote
the production of strawberry fruit. Another study reported that
red light increased the number of blossoms on rose flowers and
was also the favourable light for growth of the tomato fruit due
to the morphogenetic reaction of the photosynthetic plants to
changes of light conditions (Sukenik, 1991). In the present paper
the influence of light source on two strains of microalgae; Chlorella
vulgaris (green algae) and Gloeothece membranacea (cyanobacteria
or blue-green algae); by modifying light wavelength range using
luminescent LUMINSCENT sheets has been studied. The strains
were chosen as geographically local representatives of both pro-
karyotic and eukaryotic microalgae. The choice of local species
minimises any environmental impact in the event of accidental re- Fig. 1. Graph of light intensity (counts) vs. light wavelength (nm) for different
luminescent-filtered xenon light as provided for algal growth. Unfiltered, control
lease from large-scale commercial culture. The study investigates (clear PMMA), violet, green, orange and red filters show different wavelength
the effects of the selected light source on biomass content and pho- spectra. (For interpretation of the references to colour in this figure legend, the
tosynthetic pigment production. All aspects of the experiments reader is referred to the web version of this article.)
 S.F. Mohsenpour et al. / Bioresource Technology 125 (2012) 75–81
Light intensity was measured in unit of photon flux (lmol m2 -
1
s ) using a PAR light meter (Skye Instruments, Quantum sensor,
Powys, Wales). Visible-emitting organic dyes were selected for this
study. The following colorants were tested and their emission
spectrum was measured:
– Lumogen F Violet 570 (naphthalimide)
– Lumogen F Yellow 083 (perylene)
– Lumogen F Orange 240 (perylene)
– Lumogen F Red 305 (perylene)
– Clear (no colourant; used as control growth conditions in all
cases).
The fluorescent dyes were from Lumogen F dyes series (BASF
Aktiengesellschaft, Ludwigshafen, Germany). Luminescent acrylic
sheets containing different dyes were placed as filters directly be-
tween the xenon lamps and the culture flasks such that the light
transmitted and emitted through the face of the sheets became
incident on the culture flasks. Fig. 2 illustrates cross section of a
luminescent sheet showing different light paths when illuminated
by a light source. Photons of light can be absorbed by the host
material (PMMA) and the fluorophores. Small amount of photons
can also be reflected through the surface. However, the majority
of the photons are transmitted through the surface and the edges
of the luminescent sheet. Depending on the wavelength range of
fluorophores, the photons of light absorbed by fluorophores were
emitted at a different wavelength and received by algae culture.
2.3. Cultivation condition
The seed cultures of both microalgae strains were prepared by
cultivating in a 250 mL flask containing 100 mL of media (3N-
BBM+V for green algae and BG 11 for cyanobacteria). The cultures
were incubated in a light box at 20 °C for a week under cool white
fluorescent lighting with an intensity of approximately 20 lmol
photons m2 s1 on 12:12 h light: dark period. 20 mL of the seed
cultures were then used for inoculation of the main growth media.
Optical densities of the new cultures of C. vulgaris and G. membran-
acea after inoculations were 1.6 and 1.0, respectively. The cultures
were transferred to light boxes. The light boxes were designed as
rectangular chambers illuminated by full spectrum xenon lamps
using luminescent sheets as light filters and ventilation system to
prevent overheating inside the light box. The temperature inside
the light box was maintained at 23 ± 2 °C and recorded twice a
day using a thermometer located near the culture flasks. Light
Fig. 2. Cross-section of a luminescent sheet showing different paths of light: (1)
incident light, (2) surface reflection, (3) light absorption and emission by fluoro-
phore molecule, (4) complete light absorption by fluorophore, (5) light absorption
by host material, (6) trapped emission, (7) light emission from the edges, (8)
fluorescence emission from the surface, (9) unabsorbed light.
77
intensity of the sunlight measured in a cloudy day in Edinburgh
was about 200–250 lmol photons m2 s1. To obtain similar out-
door light intensity of a typical cloudy day in Scotland the cultures
of C. vulgaris were illuminated under the intensity of 250 lmol
photons m2 s1. Some cyanobacteria species adapted to the Scot-
tish climate show photo-inhibition even at low levels of light
intensity thus G. membranacea was illuminated under the intensity
of 150 lmol photons m2 s1. Light intensity was measured at the
same point between the luminescent sheet and the growth culture
for consistency.
2.4. Analytical methods
2.4.1. Biomass concentration
The biomass concentration was determined by a spectrophoto-
metric method in which the culture absorbance was measured at
wavelength of 680 nm. Cell dry weight was measured by filtering
the samples through a pre-dried and pre-weighed 25 mm What-
man filter and drying in the oven at 105 °C overnight. Finally, the
dried samples were placed in a dessicator and weighed for deter-
mination of biomass density. A calibration curve was obtained
showing the relationship between the cell dry biomass density
(Xb, g L1) and optical density (OD680):
X b ðC: v ulgarisÞ ¼ 0:5942  OD680 þ 0:0024 ð1Þ
X b ðG: membranaceaÞ ¼ 0:6922  OD680 þ 0:0021 ð2Þ
In addition cell concentration was determined using a haemo-
cytometer and optical microscope (CH2 Olympus optical Co. Ltd,
Tokyo, Japan). The dependence between cell concentration (Xc,
g L1) and the optical density (OD680) were established from the
calibration curves:
X c ðC: v ulgarisÞ ¼ 17:776  OD680 þ 0:4499 ð3Þ
X c ðG: membranaceaÞ ¼ 13:53  OD680 þ 0:1283 ð4Þ
2.4.2. Kinetic model
Microalgae growth can be modelled by a first order dynamic
equation:
dX=dt ¼ lX ð5Þ
1
where, l is the specific growth rate (day ), X is the biomass con-
centration (g L1) and t is the number of days, respectively. The spe-
cific growth rates of the cultures can be calculated using Eq. (5):
l ¼ ½lnðX t =X 0 Þ=ðt  t0 Þ ð6Þ
1 1
The biomass productivity rate (g L day ) which is also
known as linear growth rate can be estimated using Eq. (6):
P ¼ ðX t  X 0 Þ=ðt  t 0 Þ ð7Þ
2.4.3. Chlorophyll production
Chlorophyll concentration was determined using a modified
method of Jeffery and Humphrey (1975) for spectroscopic mea-
surements (Cheirsilp and Torpee, 2012). Due to cell density, 10–
20 mL of the cultures were withdrawn in opaque plastic bottles
and kept away from light and warmth until needed for filtration.
The samples were then filtered onto glass fibre Whatman filters
GF/F type, 25 mm in diameter. The filters were then folded once
with the algae inside and stored at 20 °C (at least). Chlorophyll
extraction was carried out by grinding the filters in a few millilitre
of 90% acetone in a glass homogenizer in an ice bath under low
light condition for 1 min. After grinding the chlorophyll extracts
were transferred to a graduated and stoppered centrifuge tube,
rinsed with exactly 10 mL of 90% acetone (i.e. 10 mL + dead volume
78 S.F. Mohsenpour et al. / Bioresource Technology 125 (2012) 75–81

of filter). The extract was centrifuged (Multifuge 3 S-R, Heraeus,

Hanau, Germany) for 10 min at 500g (where g is the gravitational
acceleration 9.81 m s2). After completion of centrifugation the
absorbance of the supernatant (OD) was measured at 750, 664,
647, and 630 nm against a 90% acetone blank. The concentration
of chlorophyll a, b and c was calculated according to the equations
below (Jeffrey and Humphrey, 1975):

Chlorophyll a ¼ ð11:85 ðOD664  OD750 Þ  1:54 ðOD647

 OD750 Þ  0:08ðOD630  OD750 ÞÞ V e =L V f ð8Þ

Chlorophyll b ¼ ð5:43 ðOD664  OD750 Þ þ 21:03 ðOD647

 OD750 Þ  2:66ðOD630  OD750 ÞÞ V e =L V f ð9Þ

Chlorophyll c ¼ ð1:67 ðOD664  OD750 Þ  7:60 ðOD647

 OD750 Þ þ 24:52ðOD630  OD750 ÞÞ V e =L V f ð10Þ

where: L = Cuvette light-path (cm), Ve = Extraction volume (mL),

Vf = Filtered volume (L). Concentrations are given in unit mg m3.
The OD reading at 750 nm is a correction for turbidity. Because
the optical density of the extract is very sensitive to changes in
the acetone to water proportions it was important to use a 90%
acetone.

3. Results and discussions

3.1. Biomass production

3.1.1. C. vulgaris
Four different colours of luminescent acrylic sheets were used
as light filters against 150 W xenon lamps with intensity of
250 lmol m2 s1 along with a reference condition of a clear ac-
rylic sheet with no added fluorophore. The light emission in PAR
by violet light (peak wavelength 400–450 nm) produced the most
efficient wavelength bands required for growth of microalgae cells.
The highest biomass accumulation was achieved under this light
colour. However, maximum specific growth rate of 0.13 day1
was obtained under orange light (peak wavelength 585–620 nm)
at the first day of cultivation. The growth parameters are presented
in Table 1.
The biomass density versus time profiles in Fig. 3(a) shows the
growth of C. vulgaris in five different light conditions. C. vulgaris
biomass concentration increased rapidly under orange and green
light condition at the first day with specific growth rates of 0.13
and 0.10 day1, respectively however cells under violet and control
light stayed in the lag phase for two days. When exposed to red
light chlorella cells entered the exponential growth phase after
4 days.
Overall growth patterns were similar under green, orange and
violet light with slightly higher average biomass production under
green light. In addition, maximum biomass productivity of
0.03 g L1 day1 was achieved under these three light conditions
suggesting these three light conditions to favour biomass produc-
tion. However, significantly lower specific growth rate of 0.07
Fig. 3. Biomass density (dry weight) in cultures of C. vulgaris (a) and G. membran-
acea (b) under violet, green, orange, red and control light recorded over a 14 day
period. The curves are fitted using a standard sigmoid Weibull growth model. Data
are the means ± SD of 5 replicates. (For interpretation of the references to colour in
this figure legend, the reader is referred to the web version of this article.)
and 0.05 day1 were obtained under red light and control (full
spectrum) light. Red light with a narrow spectrum of 600 and
700 nm did not promote rapid growth of C. vulgaris cells. The re-
sults show that the absence of violet and green wavelength bands
had a high impact on cells productivity.
On the other hand, data obtained from the control light condi-
tion showed low biomass productivity and linear growth phase
for 13 days which suggest that shorter wavelength radiation was
a growth inhibition factor. Therefore, two light conditions which
were unfavourable for C. vulgaris cultivation affected the growth
in different ways. Lack of essential wavelength bands in PAR range
in the former resulted slow growth and low productivity whereas

Table 1
Growth parameters of C. vulgaris and G. membranacea under different light conditions.

Light condition C. vulgaris G. membranacea

C14day (106 cell mL1) lmax (day1) Productivity (g L1 day1) C14day (106 cells mL1) lmax (day1) Productivity (g L1 day1)
Control 37.48 0.06 0.02 24.78 0.15 0.04
Violet 43.29 0.11 0.03 27.66 0.16 0.05
Green 42.71 0.10 0.03 23.61 0.11 0.03
Orange 42.27 0.13 0.03 24.71 0.15 0.04
Red 35.53 0.07 0.01 15.69 0.06 0.01
 S.F. Mohsenpour et al. / Bioresource Technology 125 (2012) 75–81
in the latter presence of shorter wavelength radiation had inhibi-
tion effects on C. vulgaris cells.
3.1.2. G. membranacea
The biomass density versus time profiles in Fig. 3(b) shows the
growth of G. membranacea under five different light conditions
with the intensity of 150 lmol m2 s1. Exponential growth in cya-
nobacteria cells started at the first day of inoculation. Similar
growth patterns were obtained under all light conditions except
red light with violet light being the most efficient light and slightly
lower biomass content under orange, green and control light
conditions.
Maximum specific growth rate of 0.16 day1 was achieved at
the first day under violet light condition. Violet light resulted high-
est biomass production of 1.41 g L1 and productivity of
0.05 g L1 day1 while red light showed the least efficiency in con-
verting light energy into biomass.
Lack of essential shorter wavelength bands in red light condi-
tion highly influenced the growth of G. membranacea however
chromatic adaptation in cyanobacteria cells resulted in almost
similar growth patterns under the other four light conditions with
violet being the most efficient. High tolerance of fresh water spe-
cies of cyanobacteria cells to UV and shorter wavelength radiation
(Holzinger and Lutz, 2006) can justify its comparable growth under
control light to green and orange light.
Illumination of cyanobacteria cells by violet light could poten-
tially be the most effective way to produce maximum biomass con-
tent. The results show the great potential of use of specific and
modified wavelengths of light for improving biomass content. As
is shown in Fig. 2, the dyes used do not simply remove light of cer-
tain wavelength ranges but rather absorb light at a shorter wave-
length and re-emit at longer ones. Whilst the layout used here
does not maximise the use of re-emitted photons, Fig. 2 demon-
strates that the overall loss of luminous energy is very low, and a
significant fraction of absorbed energy is being re-emitted at a
longer wavelength. A study on the growth and oil accumulation
of marine microalgae T. suecica (Go et al., 2012) showed that the
microalgae cells remained in their exponential growth phase long-
er under lower light intensity of 108.9 lmol m2 s1 and increas-
ing light intensities did not lead to higher biomass production
due to the photo-inhibition. Since all photosynthetic pigments
(such as chlorophyll-a) have optimal absorption ranges, it seems
likely that modification of wavelength conditions to move ‘‘un-
wanted’’ wavelength regions into the absorbance regions of chloro-
phyll could improve photosynthetic performance, and hence
growth, without requiring increased light intensity. The results
presented in this paper validate that hypothesis.
Since high levels of UV radiation are harmful to microalgae
(Holzinger and Lutz, 2006) then the use of violet filter to absorb
UV radiation and re-emit the energy in the PAR range has a double
benefit of enhancing visible spectrum energy whilst limiting harm-
ful UV exposure. This is also an advantage in photobioreactor de-
sign, as glass or plastic used for construction would normally
absorb the UV energy and convert to heat. By adding the dye this
energy can pass through the photobioreactor wall as visible light.
The benefits of violet dye in this regard are demonstrated in the
improved growth of both species of microalgae when compared
to full-spectrum daylight at similar luminous intensity.
The experiments carried out using other dyes demonstrated the
variation in growth according to the photosynthetic requirements
of the two species. C. vulgaris performed well under green and or-
ange light. G. membranacea also grew well under green and orange
light but showed no real improvement in growth from daylight
equivalent in these cases. Whilst G. membranacea showed consid-
erably reduced growth under red light C. vulgaris produced an
approximately similar growth pattern under red light to that ob-
79
served under daylight equivalent. The effect of the proposed illu-
mination method was a combination of reduced UV inhibition
and increased levels of re-emitted photons. It is clearly not down
to UV inhibition alone or the clear PMMA (as control test) would
demonstrate equivalent growth levels.
3.2. Photosynthetic pigment production
3.2.1. C. vulgaris
There were three different patterns of photosynthetic pigment
production in C. vulgaris under the light conditions provided; Pig-
mentation increased considerably under red light over a period of
14 days with the highest percentages of chlorophylls per gram of
biomass. Red light induced chlorophyll-a production in particular
with specific photosynthetic pigment production of 1.29%. In
addition red light favoured the pigmentation of chlorophyll-b and
c by 0.38%, and 0.27% (g Chlorophyll/g dry biomass), respectively.
The results show that providing red wavelength bands (650–
700 nm) which is the absorption spectrum for chlorophyll-a could
potentially promote pigmentation in C. vulgaris (see Table 2).
Chlorophyll-a production under violet and full-spectrum light
followed similar profiles with linear increase for the first five days
followed by a steady trend in the next days. However, the levels of
pigmentation under these two light conditions were significantly
lower than those under orange and green light at the beginning
of the cultivation. High concentrations of chlorophyll-a in the bio-
mass under orange and green light is attributable to the high cor-
responding specific growth rates l and exponential growth phase
in the first two days. The time course of chlorophyll-a content of
C. vulgaris biomass is presented in Fig. 4(a).
3.2.2. G. membranacea
Profiles of photosynthetic pigment content of G. membranacea
biomass showed an increase in chlorophyll-a content for 3 days
under all light conditions provided with slightly higher percent-
ages under green and orange light (Fig. 4(b)).
Chlorophyll-a concentration decreased under red light gradu-
ally with time and lowest specific chlorophyll-a concentration of
0.36% was obtained. Green light showed slightly better perfor-
mance compared to orange and full-spectrum daylight and in-
duced pigmentation in cyanobacteria cells. Chlorophyll-a
concentration under violet light remained almost constant during
two weeks of cultivation. Chromatic adaptation of cyanobacteria
cells to different wavelengths of light produced the similar pat-
terns of pigmentation production under the light conditions pro-
vided in the experiments. A summary of photosynthetic pigment
contents of both microalgae strains is presented in Table 2.
Previous work in a related area has shown that red LED illumi-
nation gives the highest growth rates and chlorophyll-a production
for S. platensis (Chen et al., 2010a,b; Wang et al., 2007). This work
expands from this by using light wavelength ranges rather than
specific colours and clearly shows a species variance in terms of
optimal light wavelength ranges. We hypothesise that this
variance is at least partially due to the variation in photosynthetic
pigmentation between the species under varying light conditions.
Table 2
Maximum photosynthetic pigment content (% g Chl/g biomass) of C. vulgaris and G.
membranacea under different light conditions.
Light condition C. vulgaris G. membranacea
Chl-a Chl-b Chl-c Chl-a Chl-c
Control 0.86 0.28 0.13 1.09 0.27
Violet 0.74 0.24 0.08 1.06 0.38
Green 1.04 0.39 0.07 1.23 0.40
Orange 0.89 0.37 0.17 1.11 0.33
Red 1.29 0.38 0.27 0.94 0
80 S.F. Mohsenpour et al. / Bioresource Technology 125 (2012) 75–81
Fig. 4. Variations of chlorophyll-a content (% g-chlorophyll/g-biomass) in C. vulgaris
(a) and G. membranacea (b) biomass over time under violet, green, orange, red and
control light. Data correspond to the biomass profiles in Fig. 3(a) for 4(a) and
Fig. 3(b) for 4(b). (For interpretation of the references to colour in this figure legend,
the reader is referred to the web version of this article.)
Photosynthetic pigmentation under different light conditions
exhibited the major differences between the green algae strain C.
vulgaris and the cyanobacteria G. membranacea. Firstly, C. vulgaris
cells showed completely different behaviour when illuminated
with different light wavelengths compared to their previous light
environment. This means that for the first few days of cultivation
C. vulgaris cells were responding to the environmental alterations
by adapting their photosynthetic antenna. This resulted high chlo-
rophyll production under green, orange, and red light conditions
which contained specific wavelength bands of PAR. In comparison,
full-spectrum daylight and violet light which modified natural day-
light to PAR produced average chlorophyll contents.
G. membranacea cells on the other hand had relatively similar
pigmentation production under all light conditions. This was due
to their environmental adaption abilities enabling them to survive
with only green light or a combination of green, yellow and orange
part of light spectrum (500–650 nm) barely used by other microal-
gae species (Mur et al., 1999).
However, their average pigment concentrations were higher
than C. vulgaris cells. In both strains of microalgae the control light
condition (combination of different wavelengths of light) result an
average pigmentation value between the other light conditions.
Furthermore, there was not a linear relationship between the
biomass concentration and chlorophyll content in both of the mic-
roalgae strains which was also observed from earlier work in the
area (Sanchez Miron et al., 2002). This means that illumination of
microalgae providing specific wavelengths does not necessarily
improve both pigmentation and biomass production. The results
obtained from experiments carried out showed that although vio-
let light promoted biomass production in both species of microal-
gae it was not the most efficient light condition for inducing
pigmentation. Green light improved chlorophyll production in G.
membranacea whereas red light was the sole light condition with
significant effect on pigmentation in C. vulgaris. Therefore, the ef-
fect of light wavelengths on pigmentation was species specific.
Chlorophyll-a was the major light harvesting pigment in both
strains. However, C. vulgaris contained significant amounts of chlo-
rophyll-b and c while G. membranacea lacks chlorophyll-b. The lack
of chlorophyll-b is likely to explain the fact that unlike green algae,
cyanobacteria contain phycobilins as light harvesting pigments in a
wavelength range of 500–650 nm (Satoh et al., 2001). High pig-
mentation production under green and orange light in particular
shows that these light conditions provided the suitable wave-
lengths for G. membranacea.
In both species the growth rates and cell densities obtained are
necessarily lower than many currently used algal species. A feature
of this work has been the use of naturally-growing local species and
the mimicking of local culture conditions and under Scottish condi-
tions, algal growth is generally slow. Growth rate could likely be
increased by increased illumination intensity or by the provision
of increased concentrations of CO2 but these are not considered as
part of this specific study into illumination wavelength-related
growth behaviour. We believe that the increased growth demon-
strated is due to a combination of reduced UV inhibition and
increased levels of re-emitted photons as described. It is clearly
not down to UV inhibition alone or the clear PMMA control would
demonstrate equivalent growth levels to that shown under the fast-
est growing light conditions for each species.
The implications of this work are very significant in the area of
microalgae culture, carbon capture and photobioreactor design.
Whilst much work has been done on artificial lighting of algal cul-
ture (Ravelonandro et al., 2008), a bioreactor utilising natural light
would present the most environmentally suitable solution. In addi-
tion, it is important to mention that the irradiance supply has a di-
rect impact on the biochemical composition of microalgae cells
including lipids (Chrismadha and Borowitzka, 1994) and it can
influence the metabolism of fatty acid synthesis (Sukenik, 1991).
However, the effect of light on lipid production in microalgae is
species specific (Ho et al., 2012). Low light intensity has been re-
ported to be more suitable for lipid accumulation in two species
of microalgae (Cheirsilp and Torpee, 2012) and as a result optimal
light conditions can be different for cell growth and lipid produc-
tion. Cheirsilp and Torpee (2012) argued that lower efficiency of
oil extraction and higher cost of downstream processing were
the result of low lipid production in microalgae under high light
intensity. The contrast between the results of the study mentioned
and other reports prove the fact that those species adapted to low-
er light intensities are worth to be investigated in terms of their
valuable biochemical compositions.
In areas such as Northern Europe, where sunlight intensity is
lower and more varied throughout the year, a solution such as this
to improve the light energy to biomass conversion efficiency of lo-
cal microalgae species represents an important piece of progress.
4. Conclusions
Growth and chlorophyll-a production was significantly pro-
moted in two species of microalgae under different illumination
spectra. Luminescent acrylic sheets provided light of modified
wavelength ranges to C. vulgaris and G. membranacea. From the
 S.F. Mohsenpour et al. / Bioresource Technology 125 (2012) 75–81
data obtained highest specific growth rates of 0.131 day1 for C.
vulgaris using the orange range whilst violet range supported
growth of G. membranacea with the specific growth rate of
0.160 day1. Red light had significant effect on pigmentation of C.
vulgaris however it was the least efficient light condition for culti-
vation of G. membranacea. Modified illumination strategies such as
these could allow enhanced cultivation of microalgae under natu-
ral rather than artificial light.
Acknowledgements
The work of Ronald Millar and Curtis Davis in constructing the
light boxes is gratefully acknowledged. The regular assistance of
Eileen McEvoy in algae culture has also been invaluable. The
authors would also like to thank Dr. Arno Böhm (ColorFelx,
Germany) for provision of Lumogen dyes. The financial support
of the Scottish Funding Council as part of the Horizon Project is
gratefully acknowledged.
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