Journal of Engineering Science and Technology

Vol. XX, No. Y (Year) PPP - QQQ

© School of Engineering, Taylor’s University

MODELING THE GROWTH OF B. BRAUNII MICROALGAE

UNDER NUTRIMENT LIMITATIONS IN A PHOTOBIOREACTOR

KARIANA MORENO-SADER1, ALVARO GRACÍA-PADILLA2, JAIRO

MARTINEZ-CONSUEGRA3, JEFFREY LEÓN-PULIDO4, ÁNGEL D.
GONZÁLEZ-DELGADO1,*

1
Chemical Engineering Department, Nanomaterials and Computer Aided Process
Engineering Research Group (NIPAC), University of Cartagena, Colombia
2
Chemical Engineering Department, Particle and processes Modeling, University
of Cartagena, Colombia
3
Corporación Universitaria Minuto de Dios, Colombia
4
Chemical Engineering Department, EAN University, Colombia
*Corresponding Author: agonzalezd1@unicartagena.edu.co

Abstract
Recently, microalgae have attached great attention to producing several high-
value products, and growth kinetics seem to be necessary to predict culture
performance. This work is focused on modeling the growth of Botryococcus
braunii microalgae in a photobioreactor under the limitations of nitrogen
source. Monod model best fits experimental data, which suggested that B.
braunii microalgae did not present growth inhibition in the range of limiting
nutrient concentration (0-2 mmol/L). The modeling in a photobioreactor of 25
dm3 reported that nitrite concentration decreased 80% after 13 days, while
biomass increased from 0.3 to 0.62 g/L when initial substrate concentration is
0.15 g/L. The parameters of growth kinetic models and the modeling under
nutriment limitations provide relevant information to predict B. braunii growth,
which may be useful to increase biomass productivity in several applications.
Keywords: Microalgae, growth, kinetic models, nutrient concentration, modeling.

1
2 Moreno-Sader K. et al.

1. Introduction
In the last decades, microalgae have been widely studied as a promising source of
energy and as an alternative for wastewater treatment [1]. In addition, microalgae
show great potential in adsorption and removal of heavy metals [2] and in dyes
[3], due to their simple design and low investment in terms of initial cost and soil
requirements.
The several advantages of microalgae as high contents of lipids, proteins and
carbohydrates that can be converted to produce bio-based products [4], such as
biodiesel [5], bioethanol [6] and methane [7], or a vast diversity of products
including pigments, carotenoids, fatty acids, proteins, agricultural fertilizers,
livestock, and poultry feed [8,9]. Botryococcus braunii microalgae can be found
across the world with different characteristics and are recognized as a production
hosts of biobased materials and biofuels [10]. This green alga is considered as
one of the most outstanding candidates to produce biohydrocarbon by cultivation
[11]. The wide range of applicability of microalgae has led to the development of
growth kinetic models by considering the effect of a single limiting factor or
multiple limiting factors [12].
Many factors affect microalgae growth as light intensity, nutrient availability,
dissolved CO2 concentration, temperature, and dissolved oxygen concentration
[13]. The culture medium must contain all nutriments mandatory for cell growth,
such as nitrogen, phosphorus and, iron [14–16]. Several works have been focused
on identifying interactions between nutrients and microalgae growth. Yang et al.
[17] studied the effect of bisulfite and sulfite on B. braunii growth and reported
that bisulfite became toxic at high concentrations (>1mmol/L); however, it did
not affect the growth at concentrations below 1 mmol/L. The type of bioreactors
is also an essential aspect of microalgae growth; Ranga et al. [18] focused on the
cultivation of B. braunii in raceway and circular ponds under outdoor conditions.
They found an increase in biomass yield after 18 days of cultivation.
The mathematical models for growth kinetic of microalgae can improve
predictions of photosynthetic activity and give further insight into bioreactor
design for microalgae cultivation [19]. Hence, the objective of this research was
to model the cultivation of B. braunii microalgae considering limiting nutrient
parameters in order to predict growth performance in a photobioreactor.
2.Materials and Methods
2.1. Kinetic models of B. braunii microalgae
To model growth kinetics of Botryococcus braunii microalgae, it was required
information related to cell growth in the presence of nutrients found in the
literature. The experimental results reported by Yang et al. (2004b) were used to
represent cell growth under nitrogen source (nitrite). The concentrations of both
cells and nutrients over time were used to calculate the specific growth rate of B.
braunii. The kinetic growth models considered to assess the effects of limiting
nutrients on B. braunii growth were Monod [21], Moser [22], and Martínez-
Sancho [23]. The non- linear fitting of experimental data reported in the literature
to these models was performed through Minitab 17 computational tool.
2.2. Mathematical modeling of growth in a photobioreactor

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)
 Modeling the growth of B. Braunii microalgae… 3

Despite cellular growth is complicated and depends on several factors such as

temperature, pH, and nutrients, the governing equation of microalgae growth in a
photobioreactor is the simplified reaction described as follows:
Substrate+Cells → More cells+ Product (R 1)
The relationship between biomass formation and substrate consumption is
represented by the yield coefficient in Eq. (1), where ∆ C is the change in
concentration of biomass (C) or substrate (S).
−∆ Cc
Y c /s = (1)
∆ CS
Such a yield coefficient is required to calculate the rate of substrate
consumption (r s) given by Eq. (2). The substrate is consumed for both cellular
growth and maintenance activities; hence, two terms are included in this
expression.
rg
−r s= + mC c (2)
Y c/ s
Where r g is the growth rate of microalgae and depends on specific growth rate
and biomass concentration (C c). The parameter for maintenance activities (m ) is
found in the literature [24]. Mass balance for microalgal biomass, substrate and
the product is given by Eq. (2), (4) and (5), respectively. Where, r d is the rate of
cell death, Y p / s is the yield coefficient of product generation over substrate
consumption, and C p is the concentration of the product.

dC c
=r g−r d (3)
dt
dC s −r g
= −m Cc (4)
dt Y c/ s
dC p
=Y p/ s (−r s)(5)
dt
3. Results and Discussion
3.1 Growth models for B. braunii microalga with nitrite as limiting
nutrient
Figure 1 shows the growth curve of B. braunii microalgae, which was constructed
using the experimental results reported by Yang et al. [20]. Three different phases
can be identified from the before-mentioned curve: adaptation, exponential, and
steady phases; however, the exponential phase was considered for studying the
effects of limiting nutrients because of the low growth of this photoautotrophic
microorganism. The highest cell concentration was reached after 25 days of
culture attributed to the fact that the synthesis of energetically expensive
hydrocarbons takes longer [10]. Órpez et al. [25] obtained a similar growth curve

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)
4 Moreno-Sader K. et al.

for B. braunii microalgae in a medium of urban secondarily treated wastewater

and reported the highest concentration after 450 hours (18.75 days).
The fitting parameters of growth kinetic models are summarized in Table 1.
Monod model best fitted experimental data for B. braunii growth under nitrite
limitation as a source of nitrogen. As such model is limited in modeling
microalgae growth inhibition, the high correlation coefficient suggested that
nitrite limitations do not significantly constrain microalgae growth. This model
has been widely applied to describe the relationship between microalgae growth
and a single nutrient concentration due to its simple mathematical expression
[13].
800

700
Cells concentration (mgL-1)

600

500

400

300

200

100

0
0 5 10 15 20 25 30 35 40 45 50
Time (days)

Fig. 1. Growth curve of B. braunii microalgae in culture medium with nitrite.

Table 1. Growth kinetic models for nitrite as limiting nutrient.

Correlation
Reference Model Parameters
coefficient (R2)
Cs μmax =0.145 days−1
Monod model μ=μ max 0.897
[21] K S +C s K s =0.4824 mmol/ L
μm 1 C sn+ μ m 2 μm 1=0.127 days−1
Moser model μ= n n
[22] K S +C s μm 2=0.007 days−1 0.859
K S =0.506 mmol/ L
n=1.54
Martinez μm 1=0.144 days−1
model [23] μm 1 C s+ μ m 2 K s
μ= μm 2=−0.00088 days−1 0.879
K S +C s K S =0.47 mmol/ L

3.2. Mathematical modeling of growth in a photobioreactor

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)
 Modeling the growth of B. Braunii microalgae… 5

For modeling the growth of B. braunii under nitrite limitations, the following
assumptions were stated: i) the nutriment nitrite is the main source of nitrogen
and other sources are negligible, ii) the substrate concentration is 0.15 g/L (2
mmol/L), iii) the photobioreactor volume is 25 dm3, iv) the contributions of
temperature and pH to microalgae growth are negligible, v) the culture medium is
modified Chu 13, vi) the photobioreactor type is open pond.
The results for modeling microalgae growth in a photobioreactor are shown
in Fig. 1. The concentration of substrate (Fig. 1-a), i.e., nitrite as the primary
source of nitrogen exhibited a decrease from 0.15 g/L to 0.03 g/L after 13 days of
cultivation; hence, 80% of nitrite is consumed by the microorganism to continue
growing. These results indicated that B. braunii microalga is unable to reach the
maximum cell concentration because nitrite disappears 12 days before for the
selected initial concentration of the substrate. Fig. 1-b shows the cell growth over
time, and it was expected that biomass concentration increased from 0.3 to 0.62
g/L for 13 days because of the tendency of the growth curve for such microalgae.
The values for the rate of substrate consumption and cell growth are depicted in
Fig. 1-c) and d), which suggested that the substrate is consumed slower than cell
production. A further sensitivity analysis is needed to study the effect of the initial
concentration of nitrite on growth performance.

a) b)

c) d)

Fig. 2. Growth modeling of B. braunii in a photobioreactor

4.Conclusions
This work attempted to model the growth of B. braunii microalgae considering
limiting nutrient factors. The growth curve reveals that the highest cell

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)
6 Moreno-Sader K. et al.

concentration was reached after 25 days of cultivation, suggesting a slow growth

rate attributed to the synthesis of energetically expensive hydrocarbons. It was
also found that Monod model better described culture data for nitrite as a source
of nitrogen (R2=0.897) and its parameters were calculated as
μmax =0.145 days−1 and K s =0.4824 mmol/ L. From modeling in a
photobioreactor, the nitrite concentration decreased 80% after 13 days, while
biomass increased from 0.3 to 0.62 g/L, suggesting that the microalga is unable to
reach maximum cell concentration with an initial concentration of 0.12 g/L of
nitrite.

References
1. Eze,V.C.;Velasquez-Orta,S.B.;Hernández-García,A.;Monje-
Ramírez,I.;and Orta-Ledesma,M.T. (2018). Kinetic modelling of
microalgae cultivation for wastewater treatment and carbon dioxide
sequestration. Algal Res., 32, 131–141.
2. Mathialagan,R.;Nurlidia,M.;Shamsuddin,M.R.;and Yoshimitsu,U. (2016).
Bioremoval of lead in industrial wastewater by microalgae. J. Eng. Sci.
Technol., 11, 63–69.
3. Shanmugam,D.;and Alagappan,M. (2015). Adsorption of basic magenta
using fresh water algae and brown marine seaweed: Characterization
studies and error analysis. J. Eng. Sci. Technol., 11.
4. Awaluddin,S.A.;Izhar,S.;Yoshida,H.;DANQUAH,M.;and Harun,R.
(2016). Sub-critical water technology for enhance extraction of bioactive
compound from microalgae. J. Eng. Sci. Technol., 11, 83–92.
5. Bi,Z.;and He,B.B. (2013). Characterization of microalge for the purpose
of biofuel production. Trans. ASABE, 56 (4), 1529–1539.
6. Jambo,S.A.;Abdulla,R.;Mohd Azhar,S.H.;Marbawi,H.;Gansau,J.A.;and
Ravindra,P. (2016). A review on third generation bioethanol feedstock.
Renew. Sustain. Energy Rev., 65, 756–769.
7. Gruber-
Brunhumer,M.R.;Jerney,J.;Zohar,E.;Nussbaumer,M.;Hieger,C.;Bochman
n,G.;Schagerl,M.;Obbard,J.P.;Fuchs,W.;and Drosg,B. (2015).
Acutodesmus obliquus as a benchmark strain for evaluating methane
production from microalgae: Influence of different storage and
pretreatment methods on biogas yield. Algal Res., 12, 230–238.
8. Gong,M.;and Bassi,A. (2016). Carotenoids from microalgae: A review of
recent developments. Biotechnol. Adv.
9. Rhea,N.;Groppo,J.;and Crofcheck,C. (2017). Evaluation of Flocculation,
Sedimentation, and Filtration for Dewatering of Scenedesmus Algae.
Trans. ASABE, 60 (4), 1359–1367.
10. Gouveia,J.D.;Ruiz,J.;van den
Broek,L.A.M.;Hesselink,T.;Peters,S.;Kleinegris,D.M.M.;Smith,A.G.;van
der Veen,D.;Barbosa,M.J.;and Wijffels,R.H. (2017). Botryococcus

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)
 Modeling the growth of B. Braunii microalgae… 7

braunii strains compared for biomass productivity, hydrocarbon and

carbohydrate content. J. Biotechnol., 248, 77–86.
11. Naqvi,S.R.;Naqvi,M.;Noor,T.;Hussain,A.;Iqbal,N.;Uemura,Y.;and
Nishiyama,N. (2017). Catalytic Pyrolysis of Botryococcus Braunii
(microalgae) over Layered and Delaminated Zeolites for Aromatic
Hydrocarbon Production. Energy Procedia, 142, 381–385.
12. Lee,E.;and Zhang,Q. (2016). Integrated co-limitation kinetic model for
microalgae growth in anaerobically digested municipal sludge centrate.
Algal Res., 18, 15–24.
13. Lee,E.;Jalalizadeh,M.;and Zhang,Q. (2015). Growth kinetic models for
microalgae cultivation: A review. Algal Res., 12, 497–512.
14. Serrano Bermúdez,L.M. (2012). Estudio de cuatro cepas nativas de
microalgas para evaluar su potencial uso en la producción de biodiesel.
Univ. Nac. Colomb.
15. Ruiz,A. (2011). Puesta en marcha de un cultivo de microalgas para la
eliminación de nutrientes de un agua residual urbana previamente tratada
anaeróbicamente. Univ. Politécnica Val., 102.
16. Cajamar (2016). Cultivos De Microalgas a Gran Escala: Sistemas De
Producción 1. Introducción. ADNAgro, 14.
17. Yang,S.;Wang,J.;Cong,W.;Cai,Z.;and Ouyang,F. (2004). Effects of
bisulfite and sulfite on the microalga Botryococcus braunii. Enzyme
Microb. Technol., 35 (1), 46–50.
18. Ranga,A.;Ravishankar,G.A.;and Sarada,R. (2012). Cultivation of green
alga Botryococcus braunii in raceway, circular ponds under outdoor
conditions and its growth, hydrocarbon production. Bioresour. Technol.,
123, 528–533.
19. Straka,L.;and Rittmann,B. (2018). Light-dependent kinetic model for
microalgae experiencing photoacclimation, photodamage, and
photodamage repair. Algal Res., 31, 232–238.
20. Yang,S.;Wang,J.;Cong,W.;Cai,Z.;and Ouyang,F. (2004). Utilization of
nitrite as a nitrogen source by Botryococcus braunii. Biotechnol. Lett., 26
(3), 239–243.
21. Monod,J. (1949). The growth of bacterial cultures. Annu. Rev. M, 3 (Xl),
371–394.
22. Moser,H. (1958). The dynamics of bacterial populations maintained in
the chemostat., Carnegie Institution of Washington, Washington, DC.
23. Martínez Sancho,M.;Jiménez Castillo,J.M.;and El Yousfi,F. (1997).
Influence of phosphorus concentration on the growth kinetics and
stoichiometry of the microalga Scenedesmus obliquus. Process Biochem.,
32 (8), 657–664.
24. Fogler,S.H. (2008). Elements of Chemical Reaction Engineering, Pearson.

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)
8 Moreno-Sader K. et al.

25. Órpez,R.;Eugenia,M.;Hodaifa,G.;El,F.;Jbari,N.;and Sánchez,S. (2009).

Growth of the microalga Botryococcus braunii in secondarily treated
sewage. Desalination, 246 (1–3), 625–630.

Journal of Engineering Science and Technology Month Year, Vol.

XX(Y)