Skip to main content
SpringerLink
Log in

Bioremediation of ethanol wash by microalgae and generation of bioenergy feedstock

  • Published:
Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

The disposal of waste from industrial sources is becoming a serious problem throughout the world. To produce one liter (L) of ethanol, about 15-L Ethanol Spent Water (ESW) is discharged from the ethanol industry adversely impacting aquatic and terrestrial ecosystems. This study investigated the bioremediation of ESW in two stages, first filtration through biochar (byproduct after HTL processing of algal biomass) followed by bioremediation using algae. In the first step up to 99.4% toxic melanoidin pigment and suspended pollutants from ESW were removed. The colorless ESW was then tested for the growth of marine Parachlorella kessleri-I (PK-I), and freshwater Chlorella vulgaris (CV) and Chlorella sorokiniana-I (CS-I). Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) analysis was conducted before and after bioremediation of ESW using algae. The chemical oxygen demand (COD) 28,900 mg L−1 was reduced to 55–118 mg L−1, biological oxygen demand (BOD) 7200 mg L−1 was reduced to 28–55 mg L−1, total organic carbon (98.7%), phenol (64%), sugar (99.5%), nutrients and toxic metal concentrations were also reduced significantly after 12 days of algae-cultivation in ESW. The highest biomass (115.4 mg L−1 day−1) and lipid (9.1 mg L−1 day−1) productivity were observed in PK-1 using biochar-filtered ESW. The biodiesel properties and cetane number 48.1–63.9 in FAME studied by GC–MS were observed as per the international standard of fuels. The ESW cultivated algal biomass contained a significant amount of lipids, proteins, and carbohydrates compared to the control medium grown biomass. Thus algae-derived biochar has the potential to effectively remediate the hazardous ESW thereby facilitating the efficient growth of different algal species to generate the feedstock for biofuel applications.

Graphical abstract

Bioremediation of raw Ethanol Spent Wash (ESW). About 15-L wastewater generates to produce 1-L ethanol from Ethanol Distillery. The filtration of ESW through algae-based biochar facilitated the growth of marine and freshwater algae as a bioresources for various applications while remediation of wastewater as a circular bioeconomy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The datasets supporting the conclusions of this article are included within the article and its additional materials.

References

  • Ahmad I, Fatma Z, Yazdani SS, Kumar S (2013) DNA barcode and lipid analysis of new marine algae potential for biofuel. Algal Res 2:10–15

    Article  Google Scholar 

  • Amenorfenyo DK, Huang X, Zhang Y, Zheng Y, Zeng Q, Zhang N, Ren J, Huang Q (2019) Microalgae brewery wastewater treatment: Potentials, benefits and the challenges. Int J Environ Res Public Health 16:19–10

    Article  Google Scholar 

  • Amin M, Chetpattananondh P (2019) Biochar from extracted marine Chlorella sp. residue for high efficiency adsorption with ultrasonication to remove Cr(VI), Zn(II) and Ni(II). Bioresour Technol 289:121578

    Article  CAS  Google Scholar 

  • APHA (1999) Standard Methods for the Examination of Water and Wastewater Part 4000 INORGANIC NONMETALLIC CONSTITUENTS. American Public Health Association

  • Balat M, Balat H (2009) Recent trends in global production and utilization of bio-ethanol fuel. Appl Energy 86:2273–2282

    Article  CAS  Google Scholar 

  • Barkia I, Saari N, Manning SR (2019) Microalgae for high-value products towards human health and nutrition. Mar Drugs 17:304

  • Beltran de Heredia J, Dominguez JR, Partido E (2005) Physico-chemical treatment for the depuration of wine distillery wastewaters (vinasses). Water Sci Technol 51:159–166

    Article  CAS  Google Scholar 

  • Bird MI, Wurster CM, de Paula Silva PH, Bass AM, De Nys R (2011) Algal biochar - production and properties. Bioresour Technol 102:1886–1891

    Article  CAS  Google Scholar 

  • Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 2:911–917

    Article  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Candido C, Lombardi AT (2017) Growth of Chlorella vulgaris in treated conventional and biodigested vinasses. J Appl Phycol 29:45–53

    Article  CAS  Google Scholar 

  • Chandra R, Bharagava RN, Rai V (2008) Melanoidins as major colourant in sugarcane molasses based distillery effluent and its degradation. Bioresour Technol 99:4648–4660

    Article  CAS  Google Scholar 

  • Choix FJ, Ochoa-Becerra MA, Hsieh-Lo M, Mondragón-Cortez P, Méndez-Acosta HO (2018) High biomass production and CO2 fixation from biogas by Chlorella and Scenedesmus microalgae using tequila vinasses as culture medium. J Appl Phycol 30:2247–2258

    Article  CAS  Google Scholar 

  • Cole AJ, Paul NA, de Nys R, Roberts DA (2017) Good for sewage treatment and good for agriculture: Algal based compost and biochar. J Environ Manage 200:105–113

    Article  CAS  Google Scholar 

  • Dahiya J, Singh D, Nigam P (2001) Decolourisation of synthetic and spentwash melanoidins using the white-rot fungus Phanerochaete chrysosporium JAG-40. Bioresour Technol 78:95–98

    Article  CAS  Google Scholar 

  • Das B, Deka S (2019) A cost-effective and environmentally sustainable process for phycoremediation of oil field formation water for its safe disposal and reuse. Sci Rep 9:15232

  • Demirbaş A (1998) Fuel properties and calculation of higher heating values of vegetable oils. Fuel 77:1117–1120

    Article  Google Scholar 

  • Deviram G, Mathimani T, Anto S, Ahamed TS, Ananth DA, Pugazhendhi A (2020) Applications of microalgal and cyanobacterial biomass on a way to safe, cleaner and a sustainable environment. J Clean Prod 253:119770

    Article  CAS  Google Scholar 

  • Enaime G, Baçaoui A, Yaacoubi A, Lübken M (2020) Biochar for wastewater treatment-conversion technologies and applications. Appl Sci 10:3492

    Article  CAS  Google Scholar 

  • Fabregas J, Maseda A, Domingues A, Otero A (2004) The cell composition of Nannochloropsis sp. changes under different irradiances in semicontinuous culture. World J Microbiol Biotechnol 20:31–35

    Article  CAS  Google Scholar 

  • Fito J, Tefera N, Kloos H, Van SWH (2019) Physicochemical properties of the sugar industry and ethanol distillery wastewater and their impact on the environment. Sugar Tech 21:265–277

    Article  CAS  Google Scholar 

  • Fitzgibbon FJ, Nigam P, Singh D, Marchant R (1995) Biological treatment of distillery waste for pollution-remediation. J Basic Microbiol 35:293–301

    Article  CAS  Google Scholar 

  • Gorman DS, Levine RP (1965) Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Nat Acad Sci 54:1665–1669

    Article  CAS  Google Scholar 

  • Gupta P, Ann TW, Lee SM (2016) Use of biochar to enhance constructed wetland performance in wastewater reclamation. Environ Eng Res 21:36–44

    Article  Google Scholar 

  • Hassan UM, Aamer M, Umer Chattha M, Haiying T, Khan I, Seleiman MF, Rasheed A, Nawaz M, Rehman A, Talha Aslam M, Afzal A, Huang G (2021) Sugarcane distillery spent wash (DSW) as a bio-nutrient supplement: A win-win option for sustainable crop production. Agronomy 11:183

  • Hempel N, Petrick I, Behrendt F (2012) Biomass productivity and productivity of fatty acids and amino acids of microalgae strains as key characteristics of suitability for biodiesel production. J Appl Phycol 24:1407–1418

    Article  CAS  Google Scholar 

  • Higgins BT, Thornton-Dunwoody A, Labavitch JM, Vandergheynst JS (2014) Microplate assay for quantitation of neutral lipids in extracts from microalgae. Anal Biochem 465:81–89

    Article  CAS  Google Scholar 

  • Hung CM, Huang CP, Hsieh SL, Tsai ML, Chen CW, Dong CD (2020) Biochar derived from red algae for efficient remediation of 4-nonylphenol from marine sediments. Chemosphere 254:126916

    Article  CAS  Google Scholar 

  • Jung KW, Kim K, Jeong TU, Ahn KH (2016) Influence of pyrolysis temperature on characteristics and phosphate adsorption capability of biochar derived from waste-marine macroalgae (Undaria pinnatifida roots). Bioresour Technol 200:1024–1028

    Article  CAS  Google Scholar 

  • Kharayat Y (2012) Distillery wastewater: bioremediation approaches. J Integr Environ Sci 9:69–91

    Article  Google Scholar 

  • Krisnangkura K (1986) A simple method for estimation of cetane index of vegetable oil methyl esters. J Am Oil Chem Soc 63:552–553

    Article  CAS  Google Scholar 

  • Kukreja K, Gahlot DK, Suneja S (2012) Distillery effluent: Problems and prospects-a review. Ag Rev 33:122–134

    Google Scholar 

  • Lois-Milevicich J, Casá N, Alvarez P, Mateucci R, Busto V, de Escalada PM (2020) Chlorella vulgaris biomass production using brewery wastewater with high chemical oxygen demand. J Appl Phycol 32:2773–2783

  • Lutzu GA, Zhang W, Liu T (2016) Feasibility of using brewery wastewater for biodiesel production and nutrient removal by Scenedesmus dimorphus. Environ Technol 37:1568–1581

    Article  CAS  Google Scholar 

  • Madadi R, Maljaee H, Serafim LS, Ventura SPM (2021) Microalgae as contributors to produce biopolymers. Mar Drugs 19:466

    Article  CAS  Google Scholar 

  • Michalak I, Baśladyńska S, Mokrzycki J, Rutkowski P (2019) Biochar from a freshwater macroalga as a potential biosorbent for wastewater treatment. Water (Basel) 11:4–6

  • Mikucka W, Zielińska M (2020) Distillery stillage: characteristics, treatment, and valorization. Appl Biochem Biotechnol 192:770–793

    Article  CAS  Google Scholar 

  • Molina-Cortés A, Sánchez-Motta T, Tobar-Tosse F, Quimbaya M (2020) Spectrophotometric estimation of total phenolic content and antioxidant capacity of molasses and vinasses generated from the sugarcane industry. Waste Biomass Valoriz 11:3453–3463

    Article  Google Scholar 

  • Morris I, Glover HE, Yentsch CS (1974) Products of photosynthesis by marine phytoplankton: the effect of environmental factors on the relative rates of protein synthesis. Mar Biol 27:1–9

    Article  CAS  Google Scholar 

  • Motsara MR, Roy RN (2008) Guide to laboratory establishment for plant nutrient analysis FAO fertilizer and plant nutrition bulletin. Food and Agriculture Organization, Rome

    Google Scholar 

  • Nascimento IA, Marques SSI, Cabanelas ITD, Pereira SA, Druzian JI, de Souza CO, Vich DV, de Carvalho GC, Nascimento MA (2013) Screening microalgae strains for biodiesel production: Lipid productivity and estimation of fuel quality based on fatty acids profiles as selective criteria. BioEnergy Res 6:1–13

  • Nautiyal P, Subramanian KA, Dastidar MG (2016) Adsorptive removal of dye using biochar derived from residual algae after in-situ transesterification: Alternate use of waste of biodiesel industry. J Environ Manage 182:187–197

    Article  CAS  Google Scholar 

  • Nawkarkar P, Singh AK, Abdin MZ, Kumar S (2019) Life cycle assessment of Chlorella species producing biodiesel and remediating wastewater. J Biosci 44:1–15

    Article  CAS  Google Scholar 

  • Nichols HW, Bold HC (1965) Trichosarcina polymorpha gen. et sp. nov. J Phycol 1:34–38

    Article  Google Scholar 

  • Niu Q, Luo J, Xia Y, Sun S, Chen Q (2017) Surface modification of bio-char by dielectric barrier discharge plasma for Hg0 removal. Fuel Process Technol 156:310–316

    Article  CAS  Google Scholar 

  • Pacheco D, Rocha AC, Pereira L, Verdelhos T (2020) Microalgae water bioremediation: Trends and hot topics. Appl Sci 20:1886

    Article  Google Scholar 

  • Park JY, Kim DK, Lee JP, Park SC, Kim YJ, Lee JS (2008) Blending effects of biodiesels on oxidation stability and low temperature flow properties. Bioresour Technol 99:1196–1203

    Article  CAS  Google Scholar 

  • Qiu R, Gao S, Lopez PA, Ogden KL (2017) Effects of pH on cell growth, lipid production and CO2 addition of microalgae Chlorella sorokiniana. Algal Res 28:192–199

    Article  Google Scholar 

  • Quintero-Dallos V, García-Martínez JB, Contreras-Ropero JE, Barajas-Solano AF, Barajas-Ferrerira C, Lavecchia R, Zuorro A (2019) Vinasse as a sustainable medium for the production of Chlorella vulgaris UTEX 1803. Water (Basel) 11:1–14

  • Rahimi M, Jazini M (2021) Mixotrophic cultivation of Chromochloris zofingiensis on glycerol, acetate, and vinasse. J Appl Phycol 33:3579–3590

    Article  CAS  Google Scholar 

  • Rai LC, Mallick N, Singh JB, Kumar HD (1991) Physiological and biochemical characteristics of a copper tolerant and a wild type strain of Anabaena doliolum under copper stress. J Plant Physiol 138:68–74

    Article  CAS  Google Scholar 

  • Ramírez-Verduzco LF, Rodríguez-Rodríguez JE, Jaramillo-Jacob ADR (2012) Predicting cetane number, kinematic viscosity, density and higher heating value of biodiesel from its fatty acid methyl ester composition. Fuel 91:102–111

    Article  Google Scholar 

  • Ramos MJ, Fernández CM, Casas A, Rodríguez L, Pérez Á (2009) Influence of fatty acid composition of raw materials on biodiesel properties. Bioresour Technol 100:261–268

    Article  CAS  Google Scholar 

  • Ravikumar Y, Razack SA, Yun J, Zhang G, Zabed HM, Qi X (2021) Recent advances in microalgae-based distillery wastewater treatment. Environ Technol Innov 24:101839

    Article  CAS  Google Scholar 

  • Ruggieri L, Cadena E, Martínez-Blanco J, Gasol CM, Rieradevall J, Gabarrell X, Gea T, Sort X, Sánchez A (2009) Recovery of organic wastes in the Spanish wine industry. Technical, economic and environmental analyses of the composting process. J Clean Prod 17:830–838

    Article  Google Scholar 

  • Santhakumaran P, Kookal SK, Mathew L, Ray JG (2019) Bioprospecting of three rapid-growing freshwater green algae, promising biomass for biodiesel production. Bioenergy Res 12:680–693

    Article  CAS  Google Scholar 

  • Sarin A, Arora R, Singh NP, Sarin R, Malhotra RK, Kundu K (2009) Effect of blends of Palm-Jatropha-Pongamia biodiesels on cloud point and pour point. Energy 34:2016–2021

    Article  CAS  Google Scholar 

  • Sinha SK, Gupta A, Bharalee R (2016) Production of biodiesel from freshwater microalgae and evaluation of fuel properties based on fatty acid methyl ester profile. Biofuels 7:105–121

    Article  CAS  Google Scholar 

  • Sumanta N, Haque CI, Nishika J, Suprakash R (2014) Spectrophotometric analysis of chlorophylls and carotenoids from commonly grown fern species by using various extracting solvents. Res J Chem Sci 2231:606X

    Google Scholar 

  • Soto MF, Diaz CA, Zapata AM, Higuita JC (2021) BOD and COD removal in vinasses from sugarcane alcoholic distillation by Chlorella vulgaris: Environmental evaluation. Biochem Eng J 176:108191

  • Spennati E, Casazza AA, Converti A (2020) Winery wastewater treatment by microalgae to produce low-cost biomass for energy production purposes. Energies 13:2490

  • Tiwari S, Gaur R, Singh R (2012) Decolorization of a recalcitrant organic compound (Melanoidin) by a novel thermotolerant yeast, Candida tropicalis RG-9. BMC Biotechnol 12:30

  • Valderrama LT, Del Campo CM, Rodriguez CM, de-Bashan LE, Bashan Y, (2002) Treatment of recalcitrant wastewater from ethanol and citric acid production using the microalga Chlorella vulgaris and the macrophyte Lemna minuscula. Water Res 36:4185–4192

    Article  CAS  Google Scholar 

  • Walkley A, Black IA (1934) An examination of the method for determining soil organic matter and proposed modification of the acid titration method. Soil Sci 37:29–38

    Article  CAS  Google Scholar 

  • Wang L, Nancollas GH (2008) Calcium orthophosphates: crystallization and dissolution. Chem Rev 108:4628–4669

    Article  CAS  Google Scholar 

  • Zhang D, Feng Y, Huang H, Khunjar W, Wang ZW (2020) Recalcitrant dissolved organic nitrogen formation in thermal hydrolysis pretreatment of municipal sludge. Environ Int 138:105629

    Article  CAS  Google Scholar 

  • Zhang Q, Chang C, Bai J, Fang S, Zhuang X, Yuan Z (2018) Mutants of Scenedesmus sp. for purifying highly concentrated cellulosic ethanol wastewater and producing biomass simultaneously. J Appl Phycol 30:969–978

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Thanks to Mr. Girish HR, ICGEB, New Delhi, for assistance in GC–MS run.

Funding

This work was supported by the funding from the Department of Biotechnology (DBT), Government of India to SK.

Author information

Author notes

  1. Santhosh K. Kookal and Prachi Nawkarkar contributed equally to this work.

Authors and Affiliations

  1. International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Santhosh Kumar Kookal, Prachi Nawkarkar, Naseem A. Gaur & Shashi Kumar

Authors
  1. Santhosh Kumar Kookal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Prachi Nawkarkar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Naseem A. Gaur
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shashi Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SKK carried out experiments of the present study. PN carried out the FAME analysis. NG helped in the procurement and analysis of the ethanol spent water samples. SK conceived the research idea and supervised all experimentations of this study. SKK and PN wrote the first draft of this manuscript. SK reviewed and edited the final manuscript. All authors provided feedback about the research, analysis and manuscript preparation and writing.

Corresponding author

Correspondence to Shashi Kumar.

Ethics declarations

Competing interest

All authors declare that there is no competing interest.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 638 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kookal, S.K., Nawkarkar, P., Gaur, N.A. et al. Bioremediation of ethanol wash by microalgae and generation of bioenergy feedstock. J Appl Phycol 35, 183–194 (2023). https://doi.org/10.1007/s10811-022-02866-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-022-02866-0

Keywords

Search

Navigation