Effect of Culture Medium for

Marine Microalgae

Vijendren s/o Krishnan

Supervisor: Prof. Dr. Uemura(13 Feb 2014)

Background studies

• Allen and Arnon medium produces highercyanobacteria concentration compared to BG-11medium. (Berberoglu et al., 2008)

• Macroalgae culture requires higher concentrationof vitamins than microalgae cultures.

• Microalgae cultured on different medium showsno morphological differences.

• Artificial and enriched medium have higherconcentration than marine environment.

• Ammonium concentration higher than 25 µM istoxic to some species.

• Medium composition may affect cellularcomposition specifically lutein and fatty acid(eicosapentaenoic acid) (Seto et al., 1984; Mironet al., 2008)

Component

Stock solution

(g/L) Quantity

Final concentration

(g/L)

Macronutrient

NaNO3 15 5 0.075

NaH2PO4H2O 5 1 5x10-3

Na2SiO39H2O 30 1 0.03

Micronutrient

FeCl36H20 3.15

Na2EDTA2H2O 4.36

CuSO45H2O 9.8 1 9.8x10-3

Na2MoO42H2O 6.3 1 6.3x10-3

ZnSO47H2O 22 1 0.022

CoCl26H2O 10 1 0.01

MnCl24H2O 180 1 0.18

Vitamins

Thiamine HCl (B1) 0.2g

Biotin (H) 1 1 1x10-3

Cyanocobalamin (B12) 1 11x10-3

F2 Medium

Composition Stock concentration

(g/L)

Quantity (ml) Final concentration

(g/L)

MgSO47H2O 120 10 1.2

NaNO3 100 10 1

CaCl2 30 10 0.3

K2HPO4 10 10 0.1

Na2EDTA 3 10 0.03

H3BO3 0.6 10 6x10-3

FeSO47H2O 0.2 10 2x10-3

MnCl2 0.14 10 1.4x10-3

ZnSO47H2O 0.033 10 3.3x10-4

Co(NO3)26H2O 0.014 0.5 7x10-6

CuSO45H2O 0.004 0.5 2x10-6

Mann and Myers Mann and Myers, 1968

Component Stock solution (g/L) Quantity

(ml/L)

Final concentration

(g/L)

+Pi K2HPO43H2O

K2HPO4 anhydrous

70

53.5

5 0.35

0.2675

-Pi 1. MgSO4 (MgSO47H20) 100 10 1

2. CaCl2 (CaCl22H20) 30 10 0.3

3. NaCl 100 10 1

4. Microelement (Add in order) Stock solution (mg/L) 10

Distilled water 850 ml - -

A&A Fe-EDTA* 400 ml - 10.96 + 16.32

MnCl24H2O 900 - 7.2x10-3

MoO3 Molybdic acid (85%)

Na2MoO42H20 (99%)

90

152.75

- 7.2x10-4

1.22x10-3

ZnSO47H2O 110 - 8.8x10-4

CuSO45H2O 39.5 - 3.16x10-4

H3BO3 1430 - 0.01144

NH4VO3 (NH4+ metavanadate) 11.5 - 9.2x10-5

CoCl26H2O 20 - 1.6x10-4

Allen and Arnon Allen and Arnon, 1955

SALT Final Concentration G/L

NaCl 24.6

KCl 0.67

CaCl22H2O 1.36

MgSO47H2O 6.29

MgCl26H2O 4.66

NaHCO3 0.18

Artificial Sea Water

Medium F2 A&A M & M ASW

Cation

Ca+2 - 0.082 0.108 0.37

Co+2 2.353x10-3 3.966x10-4 1.42x10-6 -

Cu+2 2.512x10-3 8.04x10-5 5.086x10-7 -

Fe+3 0.653 1.937 4.029x10-4 -

K+ - 2.66 0.0449 0.351

Mg+2 - 0.097 0.118 1.18

Mn+2 0.05 2x10-3 6.111x10-4

Na+ 0.5672 2.16232 0.2747 9.72

NH4+ - 1.415x10-5 - -

SO4-2 0.011 3.71 0.4678 2.4496

Zn+2 5x10-3 2x10-4 7.51x10-5

Anion

BO3-2 - 0.0108 5.709x10-3

Cl- 1.31 0.7546 0.6404 17.5334

CO3-2 - - - 0.129

MoO4- - 9.52x10-4 - -

NO-3 0.0547 - 0.729 -

PO4-3 3.442x10-3 0.146 0.0545 -

SiO3-2 8.04x10-3 - - -

Composition of various medium

• Allen M. B., and Arnon D. L., Studies on Nitrogen-Fixing Blue-GreenAlgae. I. Growth and Nitrogen Fixation by Anabaena Cylindrica Lemm.Plant Physiology, 30 (1955) 366-372.

• Berberoglua H., Jayb J., and Pilona L., Effect of nutrient media onphotobiological hydrogen production by Anabaena variabilis ATCC 29413.International Journal of Hydrogen Energy. 33 (2008 ) 1172 – 1184.

• Seto A., Wang H. L., and Hesseltine C.W., Culture Conditions AffectEicosapentaenoic Acid Content of ChIarella minutissima. JAOCS. 61 (1984)892 – 894.

• Mann, J.E., and Myers, J., On pigments, growth and photosynthesis of Phaeodactylum tricornutum. Journal of Phycology, 4 (1968) 349-355.

• Miron A. S., Go´mez A. C., Camacho F. G., Grima E. M., Chisti Y.Comparative evaluation of compact photobioreactors for large-scalemonoculture of microalgae. Journal of Biotechnology, 70 (1999) 249–270.

References