Effects of Some Nutritional Factors on the Growth of Chlorella vulgaris in a Mixotrophic Cultivation

Mau Trinh-Dang *

Faculty of Biology and Environmental Sciences, University of Science and Education (University of Danang), Danang, Vietnam.

Oanh Truong Thi Kim

Faculty of Biology and Environmental Sciences, University of Science and Education (University of Danang), Danang, Vietnam.

*Author to whom correspondence should be addressed.


The microalgae Chlorella vulgaris is not only known as a source of lipid compounds for the biofuel industry but also as a potential source of biomass production for aquaculture. This study investigated the influence of some nutritional factors (organic carbon source and C:N ratio) on the growth rate of Chlorella vulgaris. Glucose, sodium acetate, and sucrose were used to check the effect of the carbon sources. Meanwhile, five different ratios of C:N (1:1, 6:1, 12:1, 18:1, and 24:1) were tested. Results showed that glucose was the most suitable source of organic carbon for the biomass development of the Chlorella vulgaris; the ratio C:N=18:1 equivalent to 52.92 mmol/L carbon and 2.94 mmol/L nitrogen is the most appropriate ratio to promote biomass increase. The maximum growth rate of C. vulgaris recorded in this study was 0.58±0.03 day-1 in treatment supplemented with glucose at ratio C:N=18:1.

Keywords: Chlorella vulgaris, mixotrophic, organic carbon, C:N ratio

How to Cite

Trinh-Dang , M., & Kim , O. T. T. (2023). Effects of Some Nutritional Factors on the Growth of Chlorella vulgaris in a Mixotrophic Cultivation. Journal of Advances in Biology & Biotechnology, 26(2), 1–8. https://doi.org/10.9734/jabb/2023/v26i2617


Download data is not yet available.


Baldisserotto C, Popovich C, Giovanardi M, Sabia A, Ferroni L, Constenla D, Leonardi P, Pancaldi S. Photosynthetic aspects and lipid profiles in the mixotrophic alga neochloris oleoabundans as useful parameters for biodiesel production. Algal Res. 2016;16:255–265.

Lowrey J, Brooks MS, McGinn PJ. Heterotrophic and mixotrophic cultivation of microalgae for biodiesel production in agricultural wastewaters and associated challenges— A critical review. J. Appl. Phycol. 2015;27:1485–1498.

Mata TM, Martins AA, Caetano NS. Microalgae for biodiesel production and other applications: A review. Renew. Sustain. Energy Rev. 2010;14(1):217–232.

Tan HL, Lam MK, Cheng YW, Lim JW, Tan IS, Foo CYH, Show PL. Heterotrophic and mixotrophic cultivation of chlorella vulgaris using chicken waste compost as nutrients source for lipid production. In IOP Conference Series: Earth and Environmental Science; IOP Publishing, 2021;721:012011.

Dickinson S, Mientus M, Frey D, Amini-Hajibashi A, Ozturk S, Shaikh F, Sengupta D, El-Halwagi MM. A review of biodiesel production from microalgae. Clean Technol. Environ. Policy 2017;19:637–668.

Richmond A, Hu Q. Handbook of microalgal culture: Applied phycology and biotechnology; John Wiley & Sons; 2013.

Ebrahimian A, Kariminia HR, Vosoughi M. Lipid production in mixotrophic cultivation of chlorella vulgaris in a mixture of primary and secondary municipal wastewater. Renew. Energy. 2014;71:502–508.

Ji Y, Hu W, Li X, Ma G, Song M, Pei H. Mixotrophic growth and biochemical analysis of chlorella vulgaris cultivated with diluted monosodium glutamate wastewater. Bioresour. Technol. 2014;152: 471–476.

Kong W, Song H, Cao Y, Yang H, Hua S, Xia C. The characteristics of biomass production, lipid accumulation and chlorophyll biosynthesis of chlorella vulgaris under mixotrophic cultivation. Afr. J. Biotechnol. 2011;10(55):11620–11630.

Pagnanelli F, Altimari P, Trabucco F, Toro L. Mixotrophic growth of chlorella vulgaris and nannochloropsis oculata: Interaction between glucose and nitrate. J. Chem. Technol. Biotechnol. 2014;89(5):652–661.

Lu L, Wang J, Yang G, Zhu B, Pan K. Heterotrophic growth and nutrient productivities of Tetraselmis chuii using glucose as a carbon source under different C/N ratios. J. Appl. Phycol. 2017;29: 15-21.

Silaban A, Bai R, Gutierrez‐Wing MT, Negulescu II, Rusch KA. Effect of organic carbon, C: N ratio and light on the growth and lipid productivity of microalgae/cyanobacteria coculture. Eng. Life Sci. 2014;14(1):47-56.

R Core Team, R. R: A language and environment for statistical computing; 2013.

Chandra R, Rohit MV, Swamy YV, Mohan SV. Regulatory function of organic carbon supplementation on biodiesel production during growth and nutrient stress phases of mixotrophic microalgae cultivation. Bioresour. Technol. 2014;165:279–287.

Heredia-Arroyo T, Wei W, Ruan R, Hu B. Mixotrophic cultivation of chlorella vulgaris and its potential application for the oil accumulation from non-sugar materials. Biomass Bioenergy 2011;35(5):2245–2253.

Yun HS, Kim YS, Yoon HS. Effect of different cultivation modes (photoautotrophic, mixotrophic, and heterotrophic) on the growth of chlorella Sp. and biocompositions. Front. Bioeng. Biotechnol. 2021;1305.

Perez-Garcia O, Escalante FM, De-Bashan LE, Bashan Y. Heterotrophic cultures of microalgae: Metabolism and potential products. Water Res. 2011; 45(1):11–36.

Tian YT, Wang X, Cui YH, Wang SK. A symbiotic yeast to enhance heterotrophic and mixotrophic cultivation of chlorella pyrenoidosa using sucrose as the carbon source. Bioprocess Biosyst. Eng. 2020; 43(12):2243–2252.

Zhan J, Rong J, Wang Q. Mixotrophic cultivation, a preferable microalgae cultivation mode for biomass/ bioenergy production, and bioremediation, advances and prospect. Int. J. Hydrog. Energy. 2017;42(12):8505–8517.

Cheirsilp B, Torpee S. Enhanced growth and lipid production of microalgae under mixotrophic culture condition: Effect of light intensity, glucose concentration and fed-batch cultivation. Bioresour. Technol. 2012;110:510–516.

Liang Y, Sarkany N, Cui Y. Biomass and lipid productivities of chlorella vulgaris under autotrophic, heterotrophic and mixotrophic growth conditions. Biotechnol. Lett. 2009;31:1043–1049.

Ward VC, Rehmann L. Fast media optimization for mixotrophic cultivation of chlorella vulgaris. Sci. Rep. 2019;9(1): 1–10.