Abstract
The enzymatic hydrolysates of the lipid-extracted microalgal biomass residues (LMBRs) from biodiesel production were evaluated as nutritional sources for the mixotrophic growth of Chlorella vulgaris and lipid production at different temperature levels and substrate concentrations. Both parameters had a significant effect on cell growth and lipid production. It was observed that C. vulgaris could grow mixotrophically in a wide range of temperatures (20∼35 °C). The optimal temperature for cell growth and lipid accumulation of the mixotrophic growth of C. vulgaris was between 25 and 30 °C. The neutral lipids of the culture at 25 °C accounted for as much as 82 % of the total lipid content in the microalga at culture day 8. Fatty acid composition analysis showed that the increase of saturated fatty acids was proportional to the increase in temperature. The maximum biomass concentration of 4.83 g/L and the maximum lipid productivity of 164 mg/L/day were obtained at an initial total sugar concentration of 10 g/L and an initial total concentration of amino acids of 1.0 g/L but decreased at lower and higher substrate concentrations. The present results show that LMBRS could be utilized by the mixotrophic growth of C. vulgaris for microalgal lipid production under the optimum temperature and substrate concentration.
Similar content being viewed by others
References
Rawat, I., Kumar, R. R., Mutanda, T., & Bux, F. (2013). Biodiesel from microalgae: a critical evaluation from laboratory to large scale production. Applied Energy, 103, 444–467.
Wang, Z., Ma, X. C., Zhou, W. G., Min, M., Cheng, Y. L., Chen, P., Shi, J., Wang, Q., Liu, Y. H., & Ruan, R. (2013). Oil crop biomass residue-based media for enhanced algal lipid production. Applied Biochemistry and Biotechnology, 171, 689–703.
Zhou, W. G., Li, Y. C., Min, M., Hu, B., Chen, P., & Ruan, R. (2011). Local bioprospecting for high-lipid producing microalgal strains to be grown on concentrated municipal wastewater for biofuel production. Bioresource Technology, 102, 6909–6919.
Nobre, B. P., Villalobos, F., Barragán, B. E., Oliveira, A. C., Batista, A. P., Marques, P. A. S. S., Mendes, R. L., Sovová, H., Palavra, A. F., & Gouvei, L. (2013). A biorefinery from Nannochloropsis sp. microalga—extraction of oils and pigments. Production of biohydrogen from the leftover biomass. Bioresource Technology, 135, 128–136.
Zheng, H. L., Gao, Z., Yin, F. W., Ji, X. J., & Huang, H. (2012). Lipid production of Chlorella vulgaris from lipid-extracted microalgal biomass residues through two-step enzymatic hydrolysis. Bioresource Technology, 117, 1–6.
Zheng, H. L., Gao, Z., Yin, F. W., Ji, X. J., & Huang, H. (2012). Effect of CO2 supply conditions on lipid production of Chlorella vulgaris from enzymatic hydrolysates of lipid-extracted microalgal biomass residues. Bioresource Technology, 126, 24–30.
Stephens, E., Ross, I. L., Mussgnug, J. H., Wagner, L. D., Borowitzka, M. A., Posten, C. O., Kruse, O., & Hankamer, B. (2010). Future prospects of microalgal biofuel production systems. Trends in Plant Science, 15(10), 554–564.
Ehimen, E. A., Sun, Z. F., Carrington, C. G., Birch, E. J., & Eaton-Rye, J. J. (2011). Anaerobic digestion of microalgae residues resulting from the biodiesel production process. Applied Energy, 88, 3454–3463.
Sheng, J., Kim, H. W., Badalamenti, J. P., Zhou, C., Sridharakrishnan, S., Krajmalnik-Brown, R., Rittmann, B. E., & Vannela, R. (2011). Effects of temperature shifts on growth rate and lipid characteristics of Synechocystis sp. PCC6803 in a bench-top photobioreactor. Bioresource Technology, 102, 11218–11225.
Jiang, Y., & Chen, F. (2000). Effects of temperature and temperature shift on docosahexaenoic acid production by the marine microalga Crypthecodinium cohnii. Journal of American Oil Chemistry Society, 77, 613–617.
Converti, A., Casazza, A. A., Ortiz, E. Y., Perego, P., & Borghi, M. D. (2009). Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production. Chemical Engineering Process, 48, 1146–1151.
Li, Y. Q., Horsman, M., Wang, B., Wu, N., & Lan, C. Q. (2008). Effects of nitrogen sources on cell growth and lipid accumulation of green alga Neochloris oleoabundans. Applied Microbiology Biotechnology, 81, 629–636.
Muge, I. H., Idil, G., & Murat, E. (2012). Optimization of carbon and nitrogen sources for biomass and lipid production by Chlorella saccharophila under heterotrophic conditions and development of Nile red fluorescence based method for quantification of its neutral lipid content. Biochemical Engineering Journal, 61, 11–19.
Zheng, H. L., Yin, J. L., Gao, Z., Huang, H., Ji, X. J., & Dou, C. (2011). Disruption of Chlorella vulgaris cells for the release of biodiesel-producing lipids: a comparison of grinding, ultrasonication, bead milling, enzymatic lysis, and microwaves. Applied Biochemistry and Biotechnology, 164, 1215–1224.
Yan, L. S., Zhang, H. M., Chen, J. W., Lin, Z. X., Jin, Q., Jia, H. H., & Huang, H. (2009). Dilute sulfuric acid cycle spray flow-through pretreatment of corn stover for enhancement of sugar recovery. Bioresource Technology, 100(5), 1803–1808.
Heinrikson, R. L., & Meredith, S. C. (1984). Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyanate. Analysis Biochemistry, 136, 65–74.
Wellburn, A. L. (1994). The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. Journal of Plant Physiology, 144, 307–313.
Chen, W., Zhang, C., Song, L., Sommerfeld, M., & Hu, Q. (2009). A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae. Journal of Microbiology Methods, 77, 41–47.
Bligh, E. G., & Dyer, W. M. (1959). A rapid method of lipid extraction and purification. Canadian Journal of Biochemistry and Physiology, 37, 911–917.
Metchalfe, L. D., & Schmitz, A. A. (1961). The rapid preparation of fatty acid esters for gas chromatographic analysis. Analysis Biochemistry, 33, 363–372.
Ren, L. J., Ji, X. J., Huang, H., Qu, L., Feng, Y., Tong, Q. Q., & Ouyang, P. K. (2010). Development of a stepwise aeration control strategy for efficient docosahexaenoic acid production by Schizochytrium sp. Applied Microbiology and Biotechnology, 87, 1649–1656.
Duncan, D. B. (1955). Multiple range and multiple F tests. Biometrics, 11, 1–42.
Abreu, A. P., Fernandes, B., Vicente, A. A., Teixeira, J., & Dragone, G. (2012). Mixotrophic cultivation of Chlorella vulgaris using industrial dairy waste as organic carbon source. Bioresource Technology, 118, 61–66.
Borowitzka, M. A. (1998). Limits to growth. In Y. S. Wong & N. F. Y. Tam (Eds.), Wastewater treatment with algae (pp. 203–226). Berlin: Springer-Verlag.
Li, T.T., Zheng, Y.B., Yu, L., Chen, S.L. (2014). Mixotrophic cultivation of a Chlorella sorokiniana strain for enhanced biomass and lipid production. Biomass and Bioenergy. http://dx.doi.org/10.1016/j.biombioe.2014.04.010.
Ji, Y., Hu, W. R., Li, X. Q., Ma, G. X., Song, M. M., & Pei, H. Y. (2014). Mixotrophic growth and biochemical analysis of Chlorella vulgaris cultivated with diluted monosodium glutamate wastewater. Bioresource Technology, 152, 471–476.
Guan, Y. X., Wang, J., & Sun, J. X. (2011). A method for determination of hexokinase activity by RP-HPLC. Wuhan University Journal of Nature Science, 16(6), 535–540.
Liu, M. S., & Hellebust, J. A. (1974). Uptake of amino acids by the marine centric diatom Cyclotella cryptica. Canadian Journal of Microbiology, 20, 1109–1118.
Perez-Garcia, O., Escalante, F. M. E., De-Bashan, L. E., & Bashan, Y. (2011). Heterotrophic cultures of microalgae: metabolism and potential products. Water Research, 45, 11–36.
Hu, Q., Sommerfeld, M., Jarvis, E., Ghirardi, M., Posewitz, M., Seibert, M., & Darzins, A. (2008). Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant Journal, 54, 621–639.
Thi, T. Y. D., Balasubramanian, S., & Jeffrey, P. O. (2011). Screening of marine microalgae for biodiesel feedstock. Biomass & Bioenergy, 35, 2534–2544.
Chiu, S. Y., Kao, C. Y., Tsai, M. T., Ong, S. C., Chen, C. H., & Lin, C. S. (2009). Lipid accumulation and CO2 utilization of Nannochloropsis oculata in response to CO2 aeration. Bioresource Technology, 100, 833–838.
Biel, K. Y., Gates, R. D., & Muscatine, L. (2007). Effects of free amino acids on the photosynthetic carbon metabolism of symbiotic dinoflagellates. Russian Journal of Plant Physiology, 54(2), 171–183.
Molina Grima, E., Sanchez Perez, J. A., Garcia Camacho, F., Fernandez Sevilla, J. M., & Acién Fernández, F. G. (1996). Productivity analysis of outdoor chemostat cultures in tubular air-lift photobioreactors. Journal of Applied Phycology, 8, 369–380.
Masojı’dek, J., Koblı’zˇek, M., & Torzillo, G. (2004). Photosynthesis in microalgae. In A. Richmond (Ed.), Handbook of microalgal culture: biotechnology and applied phycology (pp. 20–40). London: Blackwell Science Ltd.
Li, X., Hu, H. Y., & Zhang, Y. P. (2011). Growth and lipid accumulation properties of a freshwater microalga Scenedesmus sp. under different cultivation temperature. Bioresource Technology, 102, 3098–3102.
Fallen, B. D., Pantalone, V. R., Sams, C. E., Kopsell, D. A., Vaughn, S. F., & Moser, B. R. (2011). Effect of soybean oil fatty acid composition and selenium application on biodiesel properties. Journal of American Oil Chemistry Society, 88, 1019–1028.
Bello, E. I., Out, F., & Osasona, A. (2012). Cetane number of three vegetable oils, their biodiesels and blends with diesel fuel. Journal of Petroleum Technology and Alternative Fuels, 3(5), 52–57.
Miao, X. L., & Wu, Q. Y. (2006). Biodiesel production from heterotrophic microalgal oil. Bioresource Technology, 97, 841–846.
Acknowledgments
This work was financially supported by the Minnesota Legislative-Citizen Commission on Minnesota Resources (LCCMR), the National High-tech R&D Program of China (863 Program) (Grant Nos. 2012AA021205, 2012AA101800–03, 2012AA021704, 2012AA101809, and 2014AA022004), China International Cooperation Projects (Grant No. 2014DFA61040), and the Science and Technology Project of Jiangxi Provincial Department of Science and Technology (Grant No. 20142BBF60007).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ma, X., Zheng, H., Huang, H. et al. Effects of Temperature and Substrate Concentration on Lipid Production by Chlorella vulgaris from Enzymatic Hydrolysates of Lipid-Extracted Microalgal Biomass Residues (LMBRs). Appl Biochem Biotechnol 174, 1631–1650 (2014). https://doi.org/10.1007/s12010-014-1134-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-014-1134-5