Aquacultural Engineering 12 ( 1993) 183-190

A Flat-Sided Photobioreactor for Culturing Microalgae

M. Iqbal*, D. Grey, E Stepan-Sarkissian & M. W. Fowler

Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield,

UK, S10 2TN

ABSTRACT
A V-shaped flat-sided photobioreactor (working volume: 2 litre) was
designed and used to grow the microalga Porphyridium cruentum for
biopolymer production. Mixing of culture in the bioreactor was provided
by a mixture of air/CO2 supplied through sintered glass in the form of fine
bubbles from the base of the bioreactor. The advantages of this flat-sided
photobioreactor are the high surface-to-volume ratio, efficient mixing,
low shear force, low cost and absence of wall growth.

INTRODUCTION
A large number of chemicals and potential pharmaceutical agents have
been identified in microalgae. An important member of this group is the
unicellular red alga Porphyridium cruentum which produces a variety of
compounds with present and potential future uses. Amongst these are
the red pigment phycoerythrin (Gantt & Lipschultz, 1972), the precursor
of prostaglandins, arachidonic acid (Ahem et al., 1983) and a sulphated
extraeellular polysaccharide (Jones, 1962).
Two major systems have so far been used for the cultivation of
microalgae: (a) cultivation in open systems (outdoor ponds), and (b) culti-
vation in closed systems (ponds, tubes, fermenters with inserted lights).
Outdoor ponds have relatively low construction cost. However, it is
technically difficult to monitor and control cultural and environmental
conditions in an open system. It is also particularly difficult to maintain a
monoculture in the culture system (Ben-Amotz & Avron, 1980; Vonshak
et al., 1983; De Pauw et al., 1984). One approach to the solution of these
problems is the use of a closed system, which offers a greater flexibility in
the choice of the organism under study and allows better control of cell
physiology and growth, which makes these systems more suitable for
fundamental studies.
*Present address: Biotechnology and Food Research Centre, PCSIR Labs Complex,
Ferozepur Road, Lahore 54600, Pakistan.
183
Aquacultural Engineering 0144-8609/93/S06.00 - © 1993 Elsevier Science Publishers
Ltd, England. Printed in Great Britain
184 M. lqbal, D. Grey, G. Stepan-Sarkissian, M. W. Fowler

A number of closed systems have been described, which may be a

horizontal serpentine tubular reactor (Juttner, 1977; Pirt et al., 1983;
Gudin & Thepenier, 1986; Lee, 1986), a vertical tubular reactor
(Miyamoto et al., 1988), or a fermenter with inserted lights (Laing &
Jones, 1988). Most of these systems are either very costly or suffer from
significant operating limitations, or both (Weissman et al., 1988). There-
fore, a need exists for the development of a novel, low cost, photobio-
reactor design. Towards achieving this, a bioreactor was designed,
modelled on fluidized bed reactors. The following criteria were taken
into consideration while designing this bioreactor: (a) maximum surface
area exposed to light, (b) efficient mixing and (c) inexpensive construc-
tion material.

DESIGN A N D CONSTRUCTION

The general arrangement of the bioreactor is shown in Fig. 1, while the

overall details on dimensions and working volumes of the bioreactor are

Fig, 1. Diagrammatic representation of the complete assembly. The individual

components illustrated are: (1) gas-outlet; (2) condenser; (3) inoculation port; (4) extra
port for pH or temperature measurements; (5) flat-sided photobioreactor; (6) tube lights;
(7) sampling port; (8) bioreactor stand; (9) sintered glass sparger; (10) gas-inlet; (11) gas
filter; (12) gas line; (13) gas outlet flask.
 A fiat-sided photobioreactorfor culturing microalgae 18 5

presented in Fig. 2. The vessel was made from horticultural glass with a
thickness of 3 mm, having three ports on the top and one on the side.
These ports were made of Quickfit glass screwthreads (size 24, diameter
22.5 mm) which could be sealed with plastic screw caps (Quickfit size
24) fitted with a silicone rubber ring (Quickfit size 8). The whole
assembly of these ports is illustrated in Fig. 3. The top ports provide the
means of access to the interior of the vessel for probes, such as for
monitoring pH and temperature, as well as for inoculation. The side
port, attached with a pinch-clip attachment (Smart & Fowler, 1984) is for
sampling. The tubular inlet at the base of the vessel was used for aeration
through a sintered glass filter (BDH, Grade P250). Losses in fluid due to
evaporation were contained by means of a modified 25-cm distillation
condenser mounted on top of the vessel through one of the three top
Quickfit ports mentioned above. The vessels had nominal capacity of 3
litre with a working volume of 2 litre.
The stand for the support of the 2-1itre vessel consisted of two
rectangular aluminium plates (695 x 216 x 6 mm) joined together by
four columns (12 mm in diameter and 320 mm in height). The top plate

NOMINAL VOLUME= 3.0L

WORKING VOLUME = 2.0L

Fig. 2. Dimension and working volume of the flat-sides photobioreactor.

186 M. lqbal, D. Grey, G. Stepan-Sarkissian, M. W. Fowler

~ d

Fig. 3. Complete assembly and various port components used in the construction of
the flat-sided photo-bioreactor. The individual components illustrated are: (a) glass tube;
(b) plastic screw cap; (c) silicone rubber ring; (d) glass screw thread.

had a hole, shaped like the cross-section of the bioreactor. The vessel
was lowered through this opening in the stand and was fitted in with the
help of two rubber strips. The lower plate acted as a base and provided
support to the vessel.

OPERATION

Once constructed, the performance of the bioreactor and its ability to

support growth and polysaccharide production by P. c r u e n t u m was
tested. The vessel, after thorough cleaning, was filled with the ASW
medium (Jones, 1962) before sterilization. The sterilization was per-
formed in a Cabburn Autoclave (Shoeburyness, UK) at 120°C and 1-06
kg/cm 2 for 20-40 min. After autoclaving, the bioreactor was cooled to
room temperature and then inoculated with the microalga P. cruenturn
under aseptic condition. The pinch clip attachment at the side port was
filled with 95% ethanol to avoid contamination at the time of sampling.
The bioreactor was completely assembled in a controlled temperature
room as shown in Fig. 1. The vessels were illuminated continuously with
fluorescent cool white tubes at an irradiance of 75 /~E/m2/s and were
aerated with sterile air conditioning 2.5% CO2, filtered through a
Gelman micro-filter (ACRO 50, 20 ktm) at a rate of 125-1000 ml/min/
hire. The cells were harvested from the side port through the pinch-clip
attachment. The cell numbers were determined in a WSI (Weber Scien-
 A flat-sided photobioreactorfor culturing microalgae 187

tific International Ltd, Lancing, UK) counting chamber. The level of sul-
phated polysaccharide in the medium was measured by Alcian blue
reagent (Ramus, 1977).

MICROALGAL GROWTH AND BIOPOLYMER PRODUCTION

In order to provide optimum cultural conditions, the flat-sided photo-

bioreactor was first standardized with different levels of photon flux
density (PFD), inoculum density and aeration rate as suggested by
Chaumont et al. (1988). P. c r u e n t u m cells, cultured in the bioreactor,
were maintained for 21 days. Growth and polysaccharide production,
when cultures were subjected to a range of PFD, aeration rate and inocu-
lum density, are shown in Figs. 4(a), (b) and (c). The PFD of 75/~E/mZ/s
was found to be optimum both for growth and extracellular poly-
saccharide production (Fig. 4(a)) whereas in the case of inoculum
density, a higher yield of 48.19-49.86 x 109 cells/litre and biopolymer
production of 1924 mg/litre, respectively, were obtained at the initial
culture density of 1.0-1.5 x 109 cells/litre and 1.0 x 109 cells/litre (Fig.
4(c)). The aeration rate of 500 ml/min/litre was found to be optimum for
both growth and polysaccharide production (Fig. 4(b)). After the
standardization of the flat-sided photobioreactor with these culture
conditions, the overall performance of the reactor was tested in a
number of trials. Maximum yields in terms of culture density and poly-
saccharide production are shown in Table 1. The mean culture density
and extracellular polysaccharide after 21 days of growth over the five
trials were respectively found to be 56.52 x 109 cells/litre and 2596 mg/
litre.

CONCLUSION

The results obtained in this study show the potential of the flat-sided
photobioreactor for the indoor cultivation of microalga P. c r u e n t u m
cells. The design of the photobioreactor is fundamentally novel due to its
unusual 'V' shape which enhances the efficiency of mixing and provides a
smooth environment inside the bioreactor eliminating escape-corners
where cells create dead culture zones. The flat lateral surfaces provide a
large exposed area for the cells to trap light. Other advantages of the flat-
sided photobioreactor are low cost, low shear forces and lack of algal
adhesion to the walls of the bioreactor. Scaling-up of the system for
microalgal mass cultivation is under study.
188 m. lqbal, D. Grey, G. Stepan-Sarkissian, M. W. Fowler

-', :_3 --

- I __ I-2ooo~ _,, I,o ,SOD

~o ~ ~ 20 1ooo
[ ltT l !:1 I::::1 ~ooo ~ ~

D
2 o ~ ~ ~:
~ I0 5~
_~
_~ SOD ~ _~

LU LU
25 50 75 100 125 250 500 750 1000

PHOTON FLUX DENSITY( [JE ImZls ) AERATION RATE ( m l / m i n / L i t r ¢ - )

?
~J 2soo
o s -2ooo N

-IS00

ua -1000
'-' 20-

i .SOD

0.S 1.0 13 2.0 3.0

fl
INOCULUM DENSITY (ceils x l 0 / L i t r ' ~

Fig. 4. Growth (m) and extracellular polysaccharide (D) production by P. cruentum

cells cultured in flat-sided photobioreactor under a range of (a) photon flux density; (b)
aeration rate and (c)inoculum density.

TABLE 1
Maximum Growth and Extracellular Polysaccharide Production by P. cruentum in a
Number of Cultural Trials

Trial no. 1 2 3 4 5

Culture density 58-18 53.74 57-55 57.91 54-23

(cells x 109/litre)
Extracellular polysaccharide 2567 2630 2492 2710 2581
production (mg/litre)
 A flat-sidedphotobioreactorfor culturing microalgae 189

ACKNOWLEDGEMENTS

We would like to thank Miss Pamela Bond, Alan Scragg and John
Murray for their help in designing and constructing the bioreactor. One
of us (M.I.) wishes to thank the Ministry of Science and Technology,
Government of Pakistan, for the provision of a Research Studentship.

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