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Calculation of biomass capacity of Algae based on their elemental composition

Conference Paper · January 2013

DOI: 10.13031/aim.20131620717

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Abstract Number/Control ID: 1620717

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2013 ASABE Annual International Meeting Paper Page 1

 An ASABE Meeting Presentation

Paper Number: Abstract Number1620717

Calculation of biomass capacity of Algae based on their elemental composition

Jayasurya Vijayakumar1, Dr. Gary Anderson1, Dr. Stephan Gent2, Anand Rajendran1
1
Agricultural Engineering Department, SDSU, 2Mechanical Engineering Department, SDSU
Written for presentation at the
2013 ASABE Annual International Meeting
Sponsored by ASABE
Kansas City, Missouri
July 21 – 24, 2013

Abstract The limiting and critical factors for algae growth are culture medium, pH, light, aeration,

temperature, etc. Nutrients provided in right proportions are essential to grow and harvest a high

density algal biomass or to get a high yield of the desired end product. Every element in the growth

medium plays a significant role in the production of single algal cell. The culture medium includes

major and minor nutrients that are incorporated in to the growing biomass. In this study the elemental

composition of the media and algae were presented and compared for four species. Nutrient

requirement at the element level required to grow a unit biomass is developed and presented.

Keywords. Anabaena sp, Synechocystis sp, Nannochloropsis sp, Growth media, Biomass,

Stoichiometric elemental composition, Major elements, Trace elements.

2013 ASABE Annual International Meeting Paper Page 2

Introduction

Recently algae have gotten the attention of many environmentalist and engineers because of its

possible benefits to the environment like carbon dioxide sequestration. Some algae can be used as a

fertilizer for crops because of its excellent nitrogen fixing and ability solubilizing phosphorous

(Mandal. et al., 1999). Biodiesel, bioethanol and biobutanol can be produced from algae which can

replace fossil fuel and earn carbon credits (http://www.epa.gov/ncea/biofuels/basicinfo.htm, 2013).

Although carbon emissions cannot be eliminated by the use of biofuels, the carbon can be recycled by

growing algae which consume a large volume of carbon dioxide from the atmosphere (Moheimani et

al., 2006).

Algae can be grown using primary treated waste water as a source of nutrients so it helps reduce

pollution (Hammouda et al., 1995). Algae are a good source of nutrients that can be used in food

additives (Spolaore et al., 2006). Some kinds of algae contain high levels of omega 3 fatty acids that

may be beneficial to human health (Adarme-Vega et al., 2012). Hence there is an endeavor to grow

algae in open raceway pond systems and closed systems (photobioreactors) (Shen et al., 2009). The

advantages and disadvantages of both systems are discussed in many papers (Richardson et al., 2012;

Ozkan et al., 2012; Kunjapur et al., 2010; Pulz. 2001). These organisms are sensitive to physical and

chemical stimuli like limited and surplus supply of nutrients (Sharma et al., 2012) and they can store

nutrients for future use (Pedersen et al., 1996). To grow alage, it is indispensable to know the

elemental composition of the algae, so that the required nutrients for algae growth can be identified

The authors are solely responsible for the content of this meeting presentation. The presentation does not necessarily
reflect the official position of the American Society of Agricultural and Biological Engineers (ASABE), and its printing
and distribution does not constitute an endorsement of views which may be expressed. Meeting presentations are not
subject to the formal peer review process by ASABE editorial committees; therefore, they are not to be presented as
refereed publications. Citation of this work should state that it is from an ASABE meeting paper. EXAMPLE: Author’s
Last Name, Initials. 2013. Title of Presentation. ASABE Paper No. ---. St. Joseph, Mich.: ASABE. For information about
securing permission to reprint or reproduce a meeting presentation, please contact ASABE at rutter@asabe.org or 269-
932-7004 (2950 Niles Road, St. Joseph, MI 49085-9659 USA).

2013 ASABE Annual International Meeting Paper Page 3

and supplied. The elemental composition of Anabaena sp, Chlorella sp, Synechocystis sp and

Nannochloropsis sp is discussed and the amount of nutrients that should be supplied to get the desired

concentration of biomass is presented.

Methods

Biomass elemental composition

To begin with, the elemental composition of the organisms is identified. The elements from the

growth media and its composition are also identified. Then the biomass produced using this media is

calculated with respect to each element using the equation (Mandalam et al., 1998):



 %  =

   
     
 

 
∗ ∗
      
              
∗ 100 (1)

The dry weight fraction of the cell is taken as 25% of the fresh (wet) weight of the cell. (Mandalam et

al., 1998). The cell density of the Anabaena is 1.023 g/ml (Yoon et al., 2008), Chlorella is 1.01 g/ml

(Mandalam et al., 1998), Synechocytis is 1.289 g/ml (Melis, 2009) and Nannochloropsis is 0.0002

g/ml (Hoffmann et al., 2010). Anabaena sp has the elemental composition on a percent weight basis

of phosphorous, sulphur, potassium, sodium, chlorine, calcium, magnesium and silicon as 0.9 %, 0.3

%, 0.7 %, 0.2 %, 1.6 %, 0.7 %, 0.3 % and 1.9 % respectively (Krivtsov et al. 1999). The other major

and minor elements like carbon, nitrogen, hydrogen, oxygen, zinc, ferrous, copper and manganese are

44.6 %, 7.68 %, 7 %, 25 %, 0.005 %, 0.125 %, 0.003 % and 0.043 % respectively (Roger et al., 1986).

The composition of BG-11 Media in which Anabaena Sp. (El-Sayed et al., 2010) is grown is given in

Table 1.

Table 1. The Composition of BG-11 media and the total available elemental composition in the

media (El-Sayed et al., 2010)

2013 ASABE Annual International Meeting Paper Page 4

 Compounds Weight in mg/l
NaNO3 1500
K2HPO4 40
MgSO4.7H2O 75
CaCl2.2 H2O 36
Citric acid 6
Ferric Ammonium citrate 6
EDTA 1
NaCO3 20
Trace metal mix
H3BO3 2860
MnCl2 4H2O 1810
ZnSO4 7H2O 222
NaMoO4 2H2O 390
CuSO4 5H2O 79
Co(NO3)2 6H2O 49

Table 2. The individual elements in the media BG -11.

Compounds Total available (mg/l)

Sodium 452.37
Phosphorus 7.12
Sulphur 44.61
Potassium 17.93
Chlorine 657.03
Magnesium 7.31
Calcium 9.79
Nitrogen 251.76
Zinc 50.27
Copper 19.90
Manganese 502.77
Carbon 2.891
Hydrogen 240.08
Oxygen 4118.94
2013 ASABE Annual International Meeting Paper Page 5
Table 2 shows the total available nutrients in BG-11 media calculated from the composition given in

table 1. The quantities of the elements given in table 2 are calculated as presented for potassium. The

total potassium (K) available in the media is calculated from the source of potassium in the BG-11

which is the salt K2HPO4 (table 1). The atomic weight of K2HPO4 is 174. The quantity of potassium

available in BG-11 is:

#$

"= ∗ 40 = 17.93 (2)
#% 

The element potassium has an atomic weight of 39. The salt K2HPO4 has two molecules of potassium

so the atomic weight of potassium is multiplied by 2 to get the total weight of potassium in K2HPO4.

The total weight of potassium is divided by the atomic weight of the salt and multiplied by 40 mg/l of

K2HPO4 that is available in the BG-11 media (table 2). The quantities of the other elements are

calculated by the same procedure. Algal growth can be inhibited when one of the elements is depleted

from the growth medium and it is considered to be the limiting nutrient. From the table 2, it seen that

the limiting nutrient for BG-11 media is phosphorus because it has the smallest quantity available.

However, the limiting nutrient cannot be established until the quantity of the nutrient available is

compared to the composition of the organism. If the organism requires only a small amount of the

nutrient, only a small amount of the nutrient needs to be supplied. In general, nitrate and phosphate

quantities are lower than required and will limit the growth of cyanobacteria (Ernst et al., 2005).

By having the total available nutrients in the media and with the elemental composition of the algae

the amount of biomass produced can be calculated using equation 1. The biomass produced from the

nutrient potassium available in the BG-11 media is calculated in equation 3. The dry weight of the

cells is taken as 0.25 % of fresh weight. The fraction dry weight of the element (nutrient) in

Anabaena is 0.007


   0 4
2  

+,-.// 012 %  = 17.93 ∗ 333
 ∗ 3.33# ∗ ∗ 7 89: ;<== ∗ 7 >9<?@ ;<==? ∗

 3.5607 >9<?@ ;<==?2 .35A 0B= ;<==?2


100 = 1000.9 %  (3)

This biomass calculated by equation 3 on a percent basis (v/v) is converted into mg/l by multiplying
2013 ASABE Annual International Meeting Paper Page 6
the biomass on a percent basis (v/v) by the fresh or dry cell density depending on biomass units

desired (wet or dry biomass). Table 3 shows the biomass produced from the standard growth media

BG-11 using the above method. Standard BG-11 media can produce 316.4 g/l of Anabeana biomass.

By changing the quantity of each element from 1 mg/l to 10 mg/l the amount of biomass that can be

produced is tabulated in table 3 in the columns labeled 1-10. From the elemental composition of

Anabaena, the stoichiometric chemical composition is derived and is C44.6H7O25N7.68 P0.9S0.3.

Anabaena is composed of Carbohydrates: 25-30%, Protein: 43-56%, and Lipid: 4-7%

(http://www.oilgae.com/algae/comp/comp.html, 2013). From table 3, the maximum amount of

Anabaena biomass that can be produced from BG-11 is 316.4 g/l and is limited by phosphorous.

Carbon is not considered to be limiting since it is often obtained from the air or provided separately

from the growth media.

Table 3: The amount of biomass produced for different concentration of nutrients and from

BG-11 media – Anabaena sp.

Biomass concentration g/l (fresh weight)

Elt BG-11 1 2 3 4 5 6 7 8 9 10
P 316.4 44.4 88.9 133.3 177.8 222.2 266.7 311.1 355.6 400.0 444.4
S 5948.0 133.3 266.7 400.0 533.3 666.7 800.0 933.3 1066.7 1200.0 1333.3
K 1024.6 57.1 114.3 171.4 228.6 285.7 342.9 400.0 457.1 514.3 571.4
Ca 559.4 57.1 114.3 171.4 228.6 285.7 342.9 400.0 457.1 514.3 571.4
Mg 974.7 133.3 266.7 400.0 533.3 666.7 800.0 933.3 1066.7 1200.0 1333.3
N 1311.3 5.2 10.4 15.6 20.8 26.0 31.3 36.5 41.7 46.9 52.1
C 2.6 0.9 1.8 2.7 3.6 4.5 5.4 6.3 7.2 8.1 9.0
H 1371.9 5.7 11.4 17.1 22.9 28.6 34.3 40.0 45.7 51.4 57.1
O 6590.3 1.6 3.2 4.8 6.4 8.0 9.6 11.2 12.8 14.4 16.0
Na 90474.0 200.0 400.0 600.0 800.0 1000.0 1200.0 1400.0 1600.0 1800.0 2000.0
Cl 16425.8 25.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0
Zn 402160.0 8000.0 16000.0 24000.0 32000.0 40000.0 48000.0 56000.0 64000.0 72000.0 80000.0
Cu 265333.3 13333.3 26666.7 40000.0 53333.3 66666.7 80000.0 93333.3 106666.7 120000.0 133333.3
Mn 467693.0 930.2 1860.5 2790.7 3720.9 4651.2 5581.4 6511.6 7441.9 8372.1 9302.3

The same procedure is followed for the cyanobacteria Synechocytis sp. The elemental composition of

2013 ASABE Annual International Meeting Paper Page 7

this alga is: carbon (51.4%), nitrogen (11.3%), oxygen (27.5%), hydrogen (6.1%) and other elements

(3.3%) on a percent wet weight basis (Shastri et al., 2005). This species can also be grown in BG-11

medium (Rajan et al., 2010). The composition of BG-11 media and the total nutrient content available

was discussed previously. Table 4 shows the different amounts of Synechocystis biomass that can be

produced from different concentration of nutrients in the BG-11 media. The stoichiometric chemical

composition of Synechocytis sp derived from the elemental composition is C51.4H6.1O27.5N11.3. The

organism is composed of Proteins 51%, carbohydrates 19%, and lipids 10% (Shastri et al., 2005).

Table 4 shows, the amount of biomass produced from standard BG-11 media and from 1-10 mg/l of

the different nutrients being available for growth.

Table 4: The amount of biomass produced for different concentration of nutrients and BG-11

media – Synechocytis sp.

Biomass concentration g/l (fresh weight)

Elt BG-11 1 2 3 4 5 6 7 8 9 10
C 2.2 0.8 1.6 2.3 3.1 3.9 4.7 5.4 6.2 7.0 7.8
N 891.2 3.5 7.1 10.6 14.2 17.7 21.2 24.8 28.3 31.9 35.4
H 1574.3 6.6 13.1 19.7 26.2 32.8 39.3 45.9 52.5 59.0 65.6
O 5991.2 1.5 2.9 4.4 5.8 7.3 8.7 10.2 11.6 13.1 14.5
Trace elements 21443.6 12.1 24.2 36.4 48.5 60.6 72.7 84.8 97.0 109.1 121.2

Table 4 shows that the limiting nutrient for production of Synechocytis sp with BG-11 is nitrogen

with a biomass production of 891.2 g/l.

The elemental composition of the algae Chlorella is given as: carbon (60%), oxygen (21%), hydrogen

(9%), nitrogen (6.5%), phosphorus (0.8%), potassium (1.0%) magnesium (0.8%), sulphur (0.3%),

iron (0.5%), calcium (0.08%), zinc (0.005%, copper (0.004%), and manganese (0.01%) (Percent

weight basis) (Mandalam et al., 1998). Chlorella can be grown in N-8 media and the available

nutrients in N-8 are tabulated in the table 5. (Mandalam et al., 1998).

Table 5. The Composition of N-8 media. (Ramkumar K. Mandalam et al., 1998).

2013 ASABE Annual International Meeting Paper Page 8

 Compounds Weight in mg/l
KNO3 1000
KH2PO4 740
Na2HPO4.2H2O 260
CaCl2.2H2O 13
Fe.EDTA 10
FeSO4.7H2O -
MgSO4.7H2O 50
Trace metal mix
Al2(SO4)3.18H2O 3580
MnCl2.4H2O 12980
CuSO4.5H2O 1830
ZnSO4.7H2O 3200

Table 6. The total available elemental nutrients in the media.

Compounds Total available (mg/l)

Nitrogen 138.61
Phosphorus 213.95
Potassium 598.33
Magnesium 4.878
Sulphur 1113.564
Calcium 3.537
Zinc 724.73
Copper 461.16
Manganese 3605.55
Oxygen 110889.189
Hydrogen 968.618

The nutrients like nitrogen and phosphorous, when provided in excess reduces the growth rate of

algae (Aldridge at al., 1995). The nitrogen and phosphorus supplied by the N- 8 medium is 138.61

mg/l and 213.95 mg/l which results in the production of 853 g/l and 10698 g/l of Chlorella biomass.

Given that there is sufficient nitrogen to produce 853 g/l of Chlorella biomass the amount of

2013 ASABE Annual International Meeting Paper Page 9

phosphorous provided is in excess since the phosphorous provided that would produce 10698 g/l of

biomass. It is not known if the excess quantity of phosphorous would reduce the growth rate of

Chlorella. The nutrient magnesium is the limiting nutrient supplied in a quantity that can produce 244

g/l of Chlorella biomass. Different quantities of the nutrients given in table 7 ranging from 1 mg/l to

10 mg/l can produce the different amounts of biomass that are tabulated in table 7. The stoichiometric

chemical composition of Chlorella is derived from the elemental composition of algae and that is

C60H9O21N6.5 P0.8S0.08. The organism is composed of Carbohydrates: 12-17 %, Protein: 51-58%,

Lipid: 14-22%, Nucleicacid: 4-5% (http://www.oilgae.com/algae/comp/comp.html, 2013).

Table 7: The amount of biomass produced for different concentration of nutrients and N-8

media – Chlorella sp.

Biomass concentration g/l (fresh weight)

Elt N-8 1 2 3 4 5 6 7 8 9 10

P 10698 50 100 150 200 250 300 350 400 450 500
S 148475 133 267 400 533 667 800 933 1067 1200 1333
K 23933 40 80 120 160 200 240 280 320 360 400
Ca 1769 500 1000 1500 2000 2500 3000 3500 4000 4500 5000
Mg 244 50 100 150 200 250 300 350 400 450 500
N 853 6.2 12.3 18.5 24.6 30.8 36.9 43.1 49.2 55.4 61.5
H 4305 4.4 8.9 13.3 17.8 22.2 26.7 31.1 35.6 40.0 44.4
O 211218 1.9 3.8 5.7 7.6 9.5 11.4 13.3 15.2 17.1 19.0
Zn 5797840 8000 16000 24000 32000 40000 48000 56000 64000 72000 80000
Cu 4611600 10000 20000 30000 40000 50000 60000 70000 80000 90000 100000
Mn 14422200 4000 8000 12000 16000 20000 24000 28000 32000 36000 40000

Nannochloropsis is a marine algae. It is the most suitable alga to extract biofuel from because of its

high oil content, 28.7% (Gouveia and Oliveira, 2009). This alga when grown in nitrogen deficient

condition can yield approximately 50% of its biomass as oil ( Gouveia and Oliveira. 2009). The high

oil content can be deduced from the elemental composition of Nannochloropsis, between it has less

nitrogen. The carbon, hydrogen, oxygen, nitrogen and sulphur content in Nannochloropsis is stated to

be 43.3%, 6%, 25.1%, 6.4% and 22.68 % respectively (Brown et al., 2010). Table 8 describes the

2013 ASABE Annual International Meeting Paper Page 10

elemental composition of Nannochloropsis (Mohammady et al., 2005),

Table 8. The elemental composition of Nannochloropsis. (Mohammady et al., 2005).

Elements %
P 19.43
K 2.88
Na 8.45
Cl 16.83
Ca 8.44
Mg 8.44
Br 2.01
Zn 4.05
Fe 1.33
Cu 5.46

Table 9. The Composition of f/2 media. (Coutteau, 1996).

Component Mg/l
NaNO3 0.075
NaH2PO4 H2O 0.005
Na2SiO3 9H2O 0.003
Trace metal solution
FeCl3 6H2O -
Na2EDTA 2H2O -
CuSO4 5H2O 0.0098
Na2MoO4 2H2O 0.0063
ZnSO4 7H2O 0.022
CoCl2 6H2O 0.01
MnCl2 4H2O 0.180
Vitamin solution
thiamine HCl -
(vit. B1)
biotin (vit. H)
cyanocobalamin
(vit. B12)

Table 10. The total available nutrients in f/2 media.

2013 ASABE Annual International Meeting Paper Page 11

 Components Total available mg
Sodium 0.02197
Phosphorus 0.0011
Sulphur 0.0036
Chlorine
0.0659
Nitrogen 0.0123
Zinc 0.0049
Copper 0.0024
Manganese 0.05
Silicon 0.00029
Hydrogen 0.0095
Oxygen 7.6175

Guillard’s f/2 medium is used to grow Nannochloropsis (Coutteau, 1996). The concentration of

nutrients found in f/2 medium range from 1 mg/l to 10 mg/l produces Nannochloropsis biomass as

shown in the table 11. The growth medium f/2 produces little biomass. The limiting nutrient is

phosphorous and it produces only 0.002 g/l of Nannochloropsis biomass. The low biomass production

from the f/2 media may indicate that the primary function of the media is to produce a product, biooil.

Nutrient balances must also be done to determine if the nutrient supply is sufficient to produce the

maximum amount of product with biomass production being limited. The nutrients sodium and

chlorine produce the maximum amount of biomass indicating that they are supplied in excess. The

stoichiometric chemical composition of Nannocholoropsis sp is derived from the elemental

composition of Nannochloropsis and is: C43.3H6O25.1N6.4S22.68. The organism is composed of Proteins

(52%), carbohydrates (12%), lipids (28%) (Brown et al., 2010).

Table 11. The amount of biomass produced for different concentration of nutrients and

Guillard’s f2 medium – Nannochloropsis sp.

Biomass concentration g/l (fresh weight)

2013 ASABE Annual International Meeting Paper Page 12

 Elt f2 1 2 3 4 5 6 7 8 9 10
Na 0.104 4.7 9.5 14.2 18.9 23.7 28.4 33.1 37.9 42.6 47.3
P 0.002 2.1 4.2 6.3 8.4 10.5 12.5 14.6 16.7 18.8 20.9
S 0.006 1.75 3.5 5.25 7 8.75 10.5 12.25 14 15.75 17.5
Cl 0.16 2.4 4.8 7.1 9.5 11.9 14.3 16.6 19.0 21.4 23.8
N 0.08 6.3 12.5 18.8 25.0 31.3 37.5 43.8 50.0 56.3 62.5
Zn 0.05 9.9 19.8 29.6 39.5 49.4 59.3 69.1 79.0 88.9 98.8
Cu 0.02 7.3 14.7 22.0 29.3 36.7 44.0 51.3 58.7 66.0 73.3
H 0.06 6.7 13.3 20.0 26.7 33.3 40.0 46.6 53.3 60.0 66.6
O 12.14 1.6 3.2 4.8 6.4 8.0 9.6 11.2 12.7 14.3 15.9

Conclusion:

To produce large quantities of biomass from algae/cyanobacteria sufficient nutrients should be

provided. The limiting nutrient should be identified and supplemented if higher biomass

concentrations are desired. In order to enhance the continuous growth of algae/cyanobacteria, the

nutrients should be provided in the right proportions. Nutrient balances should be performed to ensure

biomass and product needs are met. Growth media nutrients should be proportioned so that many of

the nutrients are depleted for growth or product from the media at the same time.

2013 ASABE Annual International Meeting Paper Page 13

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