Skip to main content
SpringerLink
Log in

β-Carotene production and extraction: a case study of olive mill wastewater bioremediation by Rhodotorula glutinis with simultaneous carotenoid production

  • Original Article
  • Published:
Biomass Conversion and Biorefinery Aims and scope Submit manuscript

Abstract

Over the last few years, there has been an impetuous search for natural colorant to be applied in various industries, such as cosmetics, food, and textile. This is largely driven by an increasing consumer demand for formulated ingredient products that are not only considered safer than their synthetic counterparts, but also present medicinal health benefits. However, there are a limited number of commercially available soluble organic pigments. Rhodotorula glutinis yeast can be used for the biodegradation of phenols rich olive mill wastewater, which is an abundant and jeopardous polluting waste, while simultaneously producing multiple high-added value compounds, such as the natural colorant β-carotene. Herein, the protocols were firstly optimized by screening 10 different solvents while evaluating their aptitude to dissolve pure β-carotene. The results followed the trend: water ≈ methanol < ethanol < acetonitrile < acetone < dichloromethane < diethyl ether < heptane < ethyl acetate < benzene. However, due to the high toxicity of benzene and the carotenoid precipitation by heptane, ethyl acetate, diethyl ether, and dichloromethane were selected as the most promising solvents. Thus, an integrated process for the extraction, isolation, and concentration of β-carotene was proposed from Rhodotorula glutinis, cultivated in (processing) effluent. The extraction results followed the same tendency as for pure carotenoid: dichloromethane (13 ± 1 μg g–1 yeast) < diethyl ether (15 ± 2 μg g–1 yeast) < ethyl acetate (21 ± 2 μg g–1 yeast). Regarding liquid–liquid extraction, the maximum extraction efficiency was also achieved with ethyl acetate, i.e., 100 ± 28%. Overall, these results show promising findings for a cascade biorefinery downstream process, as this not only potentially reduces the burden on the environment, but also adds value to primary economic activity.

Graphical abstract

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 statement

Authors can confirm that all relevant data are included in the article and/or its supplementary information files.

References

  1. Aggoun M, Arhab R, Cornu A et al (2016) Olive mill wastewater microconstituents composition according to olive variety and extraction process. Food Chem 209:72–80. https://doi.org/10.1016/j.foodchem.2016.04.034

    Article  CAS  PubMed  Google Scholar 

  2. Khdair A Abu-Rumman G (2020) sustainable environmental management and valorization options for olive mill byproducts in the Middle East and North Africa (MENA) region Processes 8. https://doi.org/10.3390/pr8060671

  3. Foti P Romeo FV Russo N et al (2021) Olive mill wastewater as renewable raw materials to generate high added-value ingredients for agro-food industries Appl Sci 11. https://doi.org/10.3390/app11167511

  4. Zimmerman JB, Anastas PT, Erythropel HC, Leitner W (2020) Designing for a green chemistry future. Science 367:397–400. https://doi.org/10.1126/science.aay3060

    Article  CAS  PubMed  Google Scholar 

  5. Karakaya A, Laleli Y, Takaç S (2012) Development of process conditions for biodegradation of raw olive mill wastewater by Rhodotorula glutinis. Int Biodeterior Biodegradation 75:75–82. https://doi.org/10.1016/j.ibiod.2012.09.005

    Article  CAS  Google Scholar 

  6. Karakaya A Bozkoyunlu G Laleli Y Takaç S (2014) Development of pH adjustment-based operational strategy to increase total phenol removal rate in biodegradation of olive mill wastewater by Rhodotorula glutinis Desalin Water Treat. https://doi.org/10.1080/19443994.2013.823357

  7. Schneider T, Graeff-Hönninger S, French WT et al (2013) Lipid and carotenoid production by oleaginous red yeast Rhodotorula glutinis cultivated on brewery effluents. Energy 61:34–43. https://doi.org/10.1016/j.energy.2012.12.026

    Article  CAS  Google Scholar 

  8. Park PK, Kim EY, Chu KH (2007) Chemical disruption of yeast cells for the isolation of carotenoid pigments. Sep Purif Technol 53:148–152. https://doi.org/10.1016/j.seppur.2006.06.026

    Article  CAS  Google Scholar 

  9. Saha S, Walia S, Kundu A et al (2015) Optimal extraction and fingerprinting of carotenoids by accelerated solvent extraction and liquid chromatography with tandem mass spectrometry. Food Chem 177:369–375. https://doi.org/10.1016/j.foodchem.2015.01.039

    Article  CAS  PubMed  Google Scholar 

  10. Amorim-Carrilho KT, Cepeda A, Fente C, Regal P (2014) Review of methods for analysis of carotenoids. Trac Trends Anal Chem 56:49–73. https://doi.org/10.1016/j.trac.2013.12.011

    Article  CAS  Google Scholar 

  11. Hernández-Almanza A, Montanez JC, Aguilar-González MA et al (2014) Rhodotorula glutinis as source of pigments and metabolites for food industry. Food Biosci 5:64–72. https://doi.org/10.1016/j.fbio.2013.11.007

    Article  CAS  Google Scholar 

  12. de Souza Mesquita LM, Martins M, Pisani LP et al (2021) Insights on the use of alternative solvents and technologies to recover bio-based food pigments. Compr Rev Food Sci Food Saf 20:787–818. https://doi.org/10.1111/1541-4337.12685

    Article  PubMed  Google Scholar 

  13. Mussagy CU Khan S Kot AM (2021) Current developments on the application of microbial carotenoids as an alternative to synthetic pigments. Crit Rev Food Sci Nutr 0:1–15. https://doi.org/10.1080/10408398.2021.1908222

  14. Michelon M, de Matos de Borba T, da Silva Rafael R, et al (2012) Extraction of carotenoids from Phaffia rhodozyma: a comparison between different techniques of cell disruption. Food Sci Biotechnol 21:1–8. https://doi.org/10.1007/s10068-012-0001-9

    Article  CAS  Google Scholar 

  15. Bogacz-Radomska L, Harasym J (2018) β-Carotene—properties and production methods. Food Qual Saf 2:69–74. https://doi.org/10.1093/fqsafe/fyy004

    Article  CAS  Google Scholar 

  16. Marova I, Carnecka M, Halienova A et al (2012) Use of several waste substrates for carotenoid-rich yeast biomass production. J Environ Manage 95:S338–S342. https://doi.org/10.1016/j.jenvman.2011.06.018

    Article  CAS  PubMed  Google Scholar 

  17. Tinoi J, Rakariyatham N, Deming RL (2005) Simplex optimization of carotenoid production by Rhodotorula glutinis using hydrolyzed mung bean waste flour as substrate. Process Biochem 40:2551–2557. https://doi.org/10.1016/j.procbio.2004.11.005

    Article  CAS  Google Scholar 

  18. Mussagy C Coutinho J Pereira J (2021) Production of carotenoids by yeast Rhodotorula glutinis CCT-2186 and their extraction using alternative solvents

  19. Mussagy C Gonzalez-Miquel M Ebinuma V Pereira J (2022) Microbial torularhodin - a comprehensive review Crit Rev Biotechnol 1. https://doi.org/10.1080/07388551.2022.2041540

  20. Mussagy C Ebinuma V Carvalho P et al (2020) Integrative platform for the selective recovery of intracellular carotenoids and lipids from Rhodotorula glutinis CCT-2186 yeast using mixtures of bio-based solvents † Green Chem 22. https://doi.org/10.1039/d0gc02992k

  21. Saini RK, Keum Y-S (2018) Carotenoid extraction methods: a review of recent developments. Food Chem 240:90–103. https://doi.org/10.1016/j.foodchem.2017.07.099

    Article  CAS  PubMed  Google Scholar 

  22. Aksu Z, Eren AT (2007) Production of carotenoids by the isolated yeast of Rhodotorula glutinis. Biochem Eng J 35:107–113. https://doi.org/10.1016/j.bej.2007.01.004

    Article  CAS  Google Scholar 

  23. Clarke KG (2013) 11 - Downstream processing. In: Clarke KG (ed) Bioprocess Engineering. Woodhead Publishing, pp 209–234

    Chapter  Google Scholar 

  24. Mezzomo N, Maestri B, dos Santos RL et al (2011) Pink shrimp (P. brasiliensis and P. paulensis) residue: Influence of extraction method on carotenoid concentration. Talanta 85:1383–1391. https://doi.org/10.1016/j.talanta.2011.06.018

    Article  CAS  PubMed  Google Scholar 

  25. Minguez-Mosquera MI, Hornero-Mendez D (1993) Separation and quantification of the carotenoid pigments in red peppers (Capsicum annuum L.), paprika, and oleoresin by reversed-phase HPLC. J Agric Food Chem 41:1616–1620. https://doi.org/10.1021/jf00034a018

    Article  CAS  Google Scholar 

  26. Alder CM, Hayler JD, Henderson RK et al (2016) Updating and further expanding GSK{’}s solvent sustainability guide. Green Chem 18:3879–3890. https://doi.org/10.1039/C6GC00611F

    Article  CAS  Google Scholar 

  27. Byrne FP Jin S Paggiola G et al (2016) Tools and techniques for solvent selection: green solvent selection guides. Sustain Chem Process 4:7. https://doi.org/10.1186/s40508-016-0051-z

  28. Dirgha Raj Joshi Nisha Adhikari (2019) An Overview on Common Organic Solvents and Their Toxicity. J Pharm Res Int 28:1–18. https://doi.org/10.9734/jpri/2019/v28i330203

    Article  CAS  Google Scholar 

  29. Rivera S, Canela R (2012) Influence of sample processing on the analysis of carotenoids in maize. Molecules 17:11255–11268. https://doi.org/10.3390/molecules170911255

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Craft NE, Soares JH (1992) Relative solubility, stability, and absorptivity of lutein and.beta.-carotene in organic solvents. J Agric Food Chem 40:431–434. https://doi.org/10.1021/jf00015a013

    Article  CAS  Google Scholar 

  31. Prat D, Pardigon O, Flemming H-W et al (2013) Sanofi’s solvent selection guide: a step toward more sustainable processes. Org Process Res Dev 17:1517–1525. https://doi.org/10.1021/op4002565

    Article  CAS  Google Scholar 

  32. Shakeel F, Haq N, Alshehri S et al (2018) Solubility, thermodynamic properties and solute-solvent molecular interactions of luteolin in various pure solvents. J Mol Liq 255:43–50. https://doi.org/10.1016/j.molliq.2018.01.155

    Article  CAS  Google Scholar 

  33. Simpson KL, Nakayama TOM, Chichester CO (1964) Biosynthesis of yeast carotenoids. J Bacteriol 88:1688–1694. https://doi.org/10.1128/jb.88.6.1688-1694.1964

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Moliné M, Libkind D, van Broock M (2012) Production of torularhodin, torulene, and $β$-carotene by rhodotorula yeasts. In: Barredo J-L (ed) Microbial Carotenoids From Fungi: Methods and Protocols. Humana Press, Totowa, NJ, pp 275–283

  35. Moliné M, Flores MR, Libkind D et al (2010) Photoprotection by carotenoid pigments in the yeast Rhodotorula mucilaginosa: the role of torularhodin. Photochem Photobiol Sci 9:1145–1151. https://doi.org/10.1039/c0pp00009d

    Article  CAS  PubMed  Google Scholar 

  36. Sharma R, Ghoshal G (2019) Optimization of carotenoids production by Rhodotorula mucilaginosa (MTCC-1403) using agro-industrial waste in bioreactor: a statistical approach. Biotechnol Reports 25:e00407. https://doi.org/10.1016/j.btre.2019.e00407

    Article  Google Scholar 

  37. Louli V, Ragoussis N, Magoulas K (2004) Recovery of phenolic antioxidants from wine industry by-products. Bioresour Technol 92:201–208. https://doi.org/10.1016/j.biortech.2003.06.002

    Article  CAS  PubMed  Google Scholar 

  38. Prommuak C, De-Eknamkul W, Shotipruk A (2008) Extraction of flavonoids and carotenoids from Thai silk waste and antioxidant activity of extracts. Sep Purif Technol 62:444–448. https://doi.org/10.1016/j.seppur.2008.02.020

    Article  CAS  Google Scholar 

Download references

Funding

This research was funded by the ERA CoBioTech within the project Rhodolive and Slovenian Research Agency within the projects P2–0152, J2-1723, J2-2492 and NC-0013.

Author information

Authors and Affiliations

  1. Department of Catalysis and Chemical Reaction Engineering, National Institute of Chemistry, Hajdrihova 19, 1000, Ljubljana, Slovenia

    Lucija Hladnik, Filipa A. Vicente, Miha Grilc & Blaž Likozar

  2. Faculty of Chemistry and Chemical Technology, University Maribor, Smetanova ulica 17, 2000, Maribor, Slovenia

    Lucija Hladnik & Blaž Likozar

  3. National Institute of Chemistry, Hajdrihova 19, 1000, Ljubljana, Slovenia

    Filipa A. Vicente & Blaž Likozar

  4. Pulp and Paper Institute, Bogišićeva 8, 1000, Ljubljana, Slovenia

    Blaž Likozar

  5. Faculty of Polymer Technology, Ozare 19, 2380, Slovenj Gradec, Slovenia

    Blaž Likozar

Authors
  1. Lucija Hladnik
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Filipa A. Vicente
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Miha Grilc
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Blaž Likozar
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Lucija Hladnik, data curation; formal analysis; investigation; methodology; resources; validation; visualization; roles/writing—original draft; writing—review and editing. Filipa A. Vicente, conceptualization; supervision; formal analysis; writing—review and editing. Miha Grilc, writing—review and editing. Blaž Likozar, conceptualization, funding acquisition, project administration, supervision, writing—review and editing.

Corresponding authors

Correspondence to Filipa A. Vicente or Blaž Likozar.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

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 25132 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hladnik, L., Vicente, F.A., Grilc, M. et al. β-Carotene production and extraction: a case study of olive mill wastewater bioremediation by Rhodotorula glutinis with simultaneous carotenoid production. Biomass Conv. Bioref. 14, 8459–8467 (2024). https://doi.org/10.1007/s13399-022-03081-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13399-022-03081-0

Keywords

Search

Navigation