Mixed culture of Chlorella sp. and wastewater wild algae for enhanced biomass and lipid accumulation in artificial wastewater medium

Highlights

RSM is used to explore the impact of different parameter on algal growth response.

Mixed algal culture promotes algal biomass and lipid accumulation.

Optimized conditions achieve maximum productivity of algal biomass and lipid.

Abstract

The purpose of this work is to study the co-cultivation of Chlorella sp. and wastewater wild algae under different cultivation conditions (i.e. CO₂, light intensity, cultivation time, and inoculation ratio) for enhanced algal biomass and lipid productivity in wastewater medium using Response Surface Methodology (RSM). The results show that mixed cultures of Chlorella sp. and wastewater wild algae increase biomass and lipid yield. Additionally, findings indicate that CO₂, light intensity and cultivation time significantly affect algal productivity. Furthermore, CO₂ concentration and light intensity, and CO₂ concentration and algal composition, have an interactive effect on biomass productivity. Under different cultivation conditions, the response of algal biomass, cell count, and lipid productivity ranges from 2.5 to 10.2 mg/mL, 1.1 × 10⁶ to 8.2 × 10⁸ cells/mL, and 1.1 × 10¹² to 6.8 × 10¹² total fluorescent units/mL, respectively. The optimum conditions for simultaneous biomass and lipid accumulation are 3.6% of CO₂ (v/v), 160 µmol/m²/s of light intensity, 1.6/2.4 of inoculation ratio (wastewater-algae/Chlorella), and 8.3 days of cultivation time. The optimal productivity is 9.8 (g/L) for dry biomass, 8.6 E+ 08 (cells/mL) for cell count, and 6.8 E+ 12 (Total FL units per mL) for lipid yield, achieving up to four times, eight times, and seven times higher productivity compared to non-optimized conditions. Provided is a supportive methodology to improve mixed algal culture for bioenergy feedstock generation and to optimize cultivation conditions in complex wastewater environments. This work is an important step forward in the development of sustainable large-scale algae cultivation for cost-efficient generation of biofuel.

Graphical abstract

Keywords

Algal biofuels / Algal mixed cultures / Algal biomass / Algal lipid / Wastewater / Response surface methodology

References

[1] Bigogno C, Khozin-Goldberg I, Boussiba S, Vonshak A, Cohen Z (2002). Lipid and fatty acid composition of the green oleaginous alga Parietochloris incisa, the richest plant source of arachidonic acid. Phytochemistry, 60(5): 497–503

[2] Borowitzka M A (1999). Commercial production of microalgae: Ponds, tanks, tubes and fermenters. Progress in Industrial Microbiology, 35: 313–321

[3] Carley K M, Kamneva N Y, Reminga J (2004). Response Surface Methodology. Pittsburgh PA: Carnegie-Mellon Univ Pittsburgh Pa School of Computer Science

[4] 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. Bioresource Technology, 102(1): 71–81

[5] Chen G, Zhao L, Qi Y (2015). Enhancing the productivity of microalgae cultivated in wastewater toward biofuel production: A critical review. Applied Energy, 137 (1): 282–291

[6] Chen X, Wang W, Li S, Xue J, Fan L, Sheng Z, Chen Y (2010). Optimization of ultrasound-assisted extraction of Lingzhi polysaccharides using response surface methodology and its inhibitory effect on cervical cancer cells. Carbohydrate Polymers, 80(3): 944–948

[7] Chisti Y (2007). Biodiesel from microalgae. Biotechnology Advances, 25(3): 294–306

[8] Clarens A F, Resurreccion E P, White M A, Colosi L M (2010). Environmental life cycle comparison of algae to other bioenergy feedstocks. Environmental Science & Technology, 44(5): 1813–1819

[9] Cloern J, Grenz C, Vidergar-Lucas L (1996). An empirical model of the phytoplankton chlorophyll: Carbon ratio-the conversion factor between productivity and growth rate. Limnology & Oceanography, 7(43): 1313–1321

[10] Colosi L M, Zhang Y, Clarens A F, White M A (2012). Will algae produce the green? Using published life cycle assessments as a starting point for economic evaluation of future algae-to-energy systems. Biofuels, 3(2): 129–142

[11] Danielsen F, Beukema H, Burgess N D, Parish F, Brühl C A, Donald P F, Murdiyarso D, Phalan B, Reijnders L, Struebig M, Fitzherbert E B (2009). Biofuel plantations on forested lands: Double jeopardy for biodiversity and climate. Conservation Biology, 23(2): 348–358

[12] De Wit C T, Van den Bergh J P (1965). Competition between herbage plants.Netherlands Journal of Agricultural Science, 13(2): 212–221

[13] Demirbas A, Fatih Demirbas M (2011). Importance of algae oil as a source of biodiesel. Energy Conversion and Management, 52(1): 163–170

[14] Francisco E C, Neves D B, Jacob-Lopes E, Franco T T (2010). Microalgae as feedstock for biodiesel production: carbon dioxide sequestration, lipid production and biofuel quality. Journal of Chemical Technology and Biotechnology (Oxford, Oxfordshire), 85(3): 395–403

[15] Garcia N S, Bonachela J A, Martiny A C (2016). Interactions between growth-dependent changes in cell size, nutrient supply and cellular elemental stoichiometry of marine Synechococcus. ISME Journal, 10: 2715–2724

[16] Georgianna D R, Mayfield S P (2012). Exploiting diversity and synthetic biology for the production of algal biofuels. Nature, 488(7411): 329–335

[17] Gopalakrishnan K K, Detchanamoorthy S (2011). Effect of media sterilization time on penicillin G production and precursor utilization in batch fermentation. Journal of Bioprocessing & Biotechniques, 01(03): 1–4

[18] Guideline T, Guideline O (2001). OECD Guidelines for the Testing of Chemicals. Paris: Oecd/Ocde

[19] Hallenbeck P C, Grogger M, Mraz M, Veverka D (2015). The use of design of experiments and response surface methodology to optimize biomass and lipid production by the oleaginous marine green alga, Nannochloropsis gaditana in response to light intensity, inoculum size and CO₂. Bioresource Technology, 184: 161–168

[20] Ho S H, Chen C Y, Chang J S (2012). Effect of light intensity and nitrogen starvation on CO₂ fixation and lipid/carbohydrate production of an indigenous microalga Scenedesmus obliquus CNW-N. Bioresource Technology, 113: 244–252

[21] Huntley M E, Redalje D G (2007). CO₂ mitigation and renewable oil from photosynthetic microbes: A new appraisal. Mitigation and Adaptation Strategies for Global Change, 12(4): 573–608

[22] Illman A M, Scragg A H, Shales S W (2000). Increase in chlorella strains calorific values when grown in low nitrogen medium. Enzyme and Microbial Technology, 27(8): 631–635

[23] Johnson K R, Admassu W (2013). Mixed algae cultures for low cost environmental compensation in cultures grown for lipid production and wastewater remediation. Journal of Chemical Technology and Biotechnology (Oxford, Oxfordshire), 88(6): 992–998

[24] Kobayashi N, Noel E A, Barnes A, Rosenberg J, DiRusso C, Black P, Oyler G A (2013a). Rapid detection and quantification of triacylglycerol by HPLC-ELSD in Chlamydomonas reinhardtii and Chlorella strains. Lipids, 48(10): 1035–1049

[25] Kobayashi N, Noel E A, Barnes A, Watson A, Rosenberg J N, Erickson G, Oyler G A (2013b). Characterization of three Chlorella sorokiniana strains in anaerobic digested effluent from cattle manure. Bioresource Technology, 150: 377–386

[26] Leite G B, Abdelaziz A E, Hallenbeck P C (2013). Algal biofuels: Challenges and opportunities. Bioresource Technology, 145: 134–141

[27] Li Y, Qin J G (2005). Comparison of growth and lipid content in three Botryococcus braunii strains. Journal of Applied Phycology, 17(6): 551–556

[28] Liu J, Huang J, Sun Z, Zhong Y, Jiang Y, Chen F (2011). Differential lipid and fatty acid profiles of photoautotrophic and heterotrophic Chlorella zofingiensis: Assessment of algal oils for biodiesel production. Bioresource Technology, 102(1): 106–110

[29] Liu Z Y, Wang G C, Zhou B C (2008). Effect of iron on growth and lipid accumulation in Chlorella vulgaris. Bioresource Technology, 99(11): 4717–4722

[30] Mona A (2013). Sustainable algal biomass products by cultivation in wastewater flows. VTT Technology, 147: 1– 84

[31] Myers R, Montgomery D C (1995). Response Surface Methodology. New York: John Willey & Sons. Inc.

[32] Morgan K C, Kalff J (1979). Effect of light and temperature interactions on growth of Cryptomonas erosa (Cryptophyceae). Journal of Phycology, 15(2): 127–134

[33] Morris I, Glover H, Yentsch C (1974). Products of photosynthesis by marine phytoplankton: The effect of environmental factors on the relative rates of protein synthesis. Marine Biology, 27(1): 1–9

[34] Nogueira D P K, Silva A F, Araújo O Q, Chaloub R M (2015). Impact of temperature and light intensity on triacylglycerol accumulation in marine microalgae. Biomass and Bioenergy, 72: 280–287

[35] Park J B, Craggs R J, Shilton A N (2011). Wastewater treatment high rate algal ponds for biofuel production. Bioresource Technology, 102(1): 35–42

[36] Rippka R, Deruelles J, Waterbury J B, Herdman M, Stanier R Y (1979). Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Journal of General Microbiology, 111(1): 1–61

[37] Roach T, Krieger-Liszkay A (2014). Regulation of photosynthetic electron transport and photoinhibition. Current Protein & Peptide Science, 15(4): 351–362

[38] Roostaei J, Zhang Y (2017). Spatially explicit life cycle assessment: Opportunities and challenges of wastewater-based algal biofuels in the United States. Algal Research, 24: 395–402

[39] Rumin J, Bonnefond H, Saint-Jean B, Rouxel C, Sciandra A, Bernard O, Cadoret J P, Bougaran G (2015). The use of fluorescent Nile red and BODIPY for lipid measurement in microalgae. Biotechnology for Biofuels, 8(1): 42–57

[40] Scott S A, Davey M P, Dennis J S, Horst I, Howe C J, Lea-Smith D J, Smith A G (2010). Biodiesel from algae: Challenges and prospects. Current Opinion in Biotechnology, 21(3): 277–286

[41] Simionato D, Basso S, Giacometti G M, Morosinotto T (2013). Optimization of light use efficiency for biofuel production in algae. Biophysical Chemistry, 182: 71–78

[42] Singh S, Singh P (2014). Effect of CO₂ concentration on algal growth: A review. Renewable & Sustainable Energy Reviews, 38: 172–179

[43] Smith A, Morris I (1980). Pathways of carbon assimilation in phytoplankton from the Antarctic Ocean. Limnology and Oceanography, 25(5): 865–872

[44] Sterner R W, Grover J P (1998). Algal growth in warm temperate reservoirs: Kinetic examination of nitrogen, temperature, light, and other nutrients. Water Research, 32(12): 3539–3548

[45] Sun Z, Chen Y F, Du J (2016). Elevated CO₂ improves lipid accumulation by increasing carbon metabolism in Chlorella sorokiniana. Plant Biotechnology Journal, 14(2): 557–566

[46] Takagi M, Karseno T, Yoshida (2006). Effect of salt concentration on intracellular accumulation of lipids and triacylglyceride in marine microalgae Dunaliella cells. Journal of Bioscience and Bioengineering, 101(3): 223–226

[47] Takagi M, Watanabe K, Yamaberi K, Yoshida T (2000). Limited feeding of potassium nitrate for intracellular lipid and triglyceride accumulation of Nannochloris sp. UTEX LB1999. Applied Microbiology and Biotechnology, 54(1): 112–117

[48] Wahidin S, Idris A, Shaleh S R M (2013). The influence of light intensity and photoperiod on the growth and lipid content of microalgae Nannochloropsis sp. Bioresource Technology, 129: 7–11

[49] Xiong W, Li X, Xiang J, Wu Q (2008). High-density fermentation of microalga Chlorella protothecoides in bioreactor for microbio-diesel production. Applied Microbiology and Biotechnology, 78(1): 29–36

[50] Zheng H, Gao Z, Yin J, Tang X, Ji X, Huang H (2012). Harvesting of microalgae by flocculation with poly (g-glutamic acid). Bioresource Technology, 112: 212–220

Abbreviations

AW, artificial wastewater; BM, biomass; CC, cell count; FL1, filter in channel 1; HT, harvesting time; IR, inoculation ratio of wastewater algae to Chlorella vulgaris; LI, Light intensity; LC, lipid content; RSM, response surface methodology; Total FL, total fluorescent units; v/v: volume/volume percent.

Acknowledgements

This work was supported by Wayne State University Faculty Start-up Funding (#176602) and by the Wayne State University Presidential Research Enhancement Program (#171908).

RIGHTS & PERMISSIONS

2018 Higher Education Press and Springer-Verlag GmbH Germany, part of Springer Nature