Effect of Nitrogen and Carbon Sources on Growth and Lipid Production from Mixotrophic Growth of Chlorella sp. KKU-S2

Mixotrophic cultivation of the isolated freshwater microalgae Chlorella sp. KKU-S2 in batch shake flask for biomass and lipid productions, different concentration of glucose as carbon substrate, different nitrogen source and concentrations were investigated. Using 1.0g/L of NaNO3 as nitrogen source, the maximum biomass yield of 10.04g/L with biomass productivity of 1.673g/L d was obtained using 40g/L glucose, while a biomass of 7.09, 8.55 and 9.45g/L with biomass productivity of 1.182, 1.425 and 1.575g/L d were found at 20, 30 and 50g/L glucose, respectively. The maximum lipid yield of 3.99g/L with lipid productivity of 0.665g/L d was obtained when 40g/L glucose was used. Lipid yield of 1.50, 3.34 and 3.66g/L with lipid productivity of 0.250, 0.557 and 0.610g/L d were found when using the initial concentration of glucose at 20, 30 and 50g/L, respectively. Process product yield (YP/S) of 0.078, 0.119, 0.158 and 0.094 were observed when glucose concentration was 20, 30, 40 and 50 g/L, respectively. The results obtained from the study shows that mixotrophic culture of Chlorella sp. KKU-S2 is a desirable cultivation process for microbial lipid and biomass production. 





References:
[1] Ahmad, A.L., Mat Yasin, N.H., Derek, C.J.C., Lim, J.K. (2011) Microalgae as a sustainable energy source for biodiesel production: A review. Renewable and Sustainable Energy Reviews 15:584–593.
[2] Huang, G.H., Chen, F., Wei, D., Zhang, X.W., Chen, G. (2010) Biodiesel production by microalgal biotechnology. Appl Energy 87:38–46.
[3] Brennan, L., Owende, P. (2010) Biofuels from microalgae-a-review of technologies for production, processing, and extractions of biofuels and co-products. Renew Energ Rev 14:557-577.
[4] Mata, T.M., Martins, A.A., Caetano, N.S., (2010) Microalgae for biodiesel production and other applications: a review. Renew Sust Energ Rev 14: 217–232.
[5] Chen, C.-Y., Yeh, K.-L., Aisyah, R., Lee, D.-J., Chang, J.-S. (2011) Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: a critical review. Bioresour. Technol. 102 (1):71–81.
[6] Yu, H., Jia, S., Dai, Y., (2009) Growth characteristics of the cyanobacterium Nostoc flagelliforme in photoautotrophic, mixotrophic and heterotrophic cultivation. J. Appl. Phycol. 21 (1):127–133.
[7] Sun, N., Wang, Y., Li, Y.T, Huang, J-C., Chen, F. (2008) Sugar-based growth, astaxanthin accumulation and carotenogenic transcription of heterotrophic Chlorella zofingiensis (Chlorophyta). Process Biochem 43:1288–1292.
[8] Ip, P.F., Chen, F. (2005) Production of astaxanthin by the green microalga Chlorella zofingiensis in the dark. Process Biochem 40:733–738.
[9] Ogawa, T., Aiba, S. (1981) Bioenergetic analysis of mixotrophic growth in Chlorella vulgaris and Scenedesmus acutus. Biotechnol Bioeng 23:1121-1132.
[10] Heredia-Arroyo, T., Wei, W., Ruan, R., Hu, B. (2011) Mixotrophic cultivation of Chlorella vulgaris and its potential application for the oil accumulation from non-sugar materials. Biomass Bioenergy, 35:2245–2253
[11] Feng, F.Y., Yang, W., Jiang, G.Z., Xu, Y.N., Kuang, T.Y.(2005) Enhancement of fatty acid production of Chlorella sp. (Chlorophyceae) by addition of glucose and sodium thiosulphate to culture medium. Process Biochem., 40:1315–1318.
[12] Perez-Garcia, O., Escalante, F.M., de-Bashan, L.E., Bashan, Y., (2011) Heterotrophic cultures of microalgae: metabolism and potential products. Water Res. 45 (1): 11–36.
[13] Wan, M., Liu, P., Xia, J., Rosenberg, J.N., Oyler, G.A., Betenbaugh, M.J., Nie, Z., Qiu, G., (2011) The effect of mixotrophy on microalgal growth, lipid content, and expression levels of three pathway genes in Chlorella sorokiniana. Appl.Microbiol. Biotechnol. 3: 835–844.
[14] Xiong, W., Gao, C., Yan, D., Wu, C., Wu, Q., (2010) Double CO2 fixation in photosynthesis-fermentation model enhances algal lipid synthesis for biodiesel production. Bioresour. Technol. 101 (7):2287–2293.
[15] Leesing, R., Nontaso, N. (2010) Microalgal oil production by green microalgae under heterotrophic cultivation. KKU Res J 15 (9): 787-793.
[16] Miller, G.L. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31: 426–432.
[17] Kwon, D.Y. and Rhee, J.S. (1986) A Simple and rapid colorimetric method for determination of free fatty acids for lipase assay. J Am Oil Chem Soc 63:89-92.
[18] Liang, Y., Sarkany, N., Cui, Y. (2009) Biomass and lipid productivities of Chlorella vulgaris under autotrophic, heterotrophic and mixotrophic growth conditions. Biotechnol Lett 31(7):1043-1049.
[19] Ratledge C., Wynn, J.P. (2002) The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. Adv. Appl. Microbiol. 51: 1-51.
[20] Leesing, R., Kookkhunthod, S. (2011) Heterotrophic growth of Chlorella sp. KKU-S2 for lipid production using molasses as a carbon substrate. Proceedings of the International Conference on Food Engineering and Biotechnology, May 28-29, 2011, Bangkok, Thailand, pp. 87-91.