Skip to main content
SpringerLink
Log in

Production, partial purification and characterization of a proteoglycan bioemulsifier from an oleaginous yeast

  • Research Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

In this study, Meyerozyma caribbica, an indigenously isolated oleaginous yeast, produced in media containing glucose a bioemulsifier that was partially characterized as a proteoglycan based on preliminary analysis. Optimization of carbon:nitrogen (C:N) ratio revealed 30:1 as the suitable ratio for enhanced production. Apart from higher emulsification activity (E24: 70–80%), this molecule showed strong emulsion stability over a wide range of pH (2.0–9.0), salinity (0.05%—10%, w/v) and temperature (− 80 °C to + 50 °C). The current study emphasizes on the determination of critical media parameters for improved and stable bioemulsifier production coupled with partial characterization and identification of the molecule. Thus, a proteoglycan-based bioemulsifier with such a stable emulsifying property can serve as a versatile and potential component in food, cosmetics and pharmaceutical formulations.

Graphic 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
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Van Hamme JD, Singh A, Ward OP (2006) Physiological aspects. Part 1 in a series of papers devoted to surfactants in microbiology and biotechnology. Biotechnol Adv 24(6):604–620. https://doi.org/10.1016/j.biotechadv.2006.08.001

    Article  PubMed  CAS  Google Scholar 

  2. Sen R (2010) Biosurfactants: advances in experimental medicine and biology, vol 672. Springer-Verlag, New York

    Book  Google Scholar 

  3. Satpute SK, Banat IM, Dhakephalkar PK, Banpurkar AG, Chopade BA (2010) Biosurfactants, bioemulsifiers and exopolysaccharides from marine microorganisms. Biotechnol Adv 28(4):436–450. https://doi.org/10.1016/j.biotechadv.2010.02.006

    Article  PubMed  CAS  Google Scholar 

  4. Chopra J, Dineshkumar R, Bhaumik M, Dhanarajan G, Kumar R, Sen R (2016) Integrated in situ transesterification for improved biodiesel production from oleaginous yeast: a value proposition for possible industrial implication. RSC Adv 6(74):70364–70373. https://doi.org/10.1039/C6RA14003C

    Article  CAS  Google Scholar 

  5. Kumar R, Dhanarajan G, Bhaumik M, Chopra J, Sen R (2017) Performance evaluation of a yeast biorefinery as a sustainable model for co-production of biomass, bioemulsifier, lipid, biodiesel and animal-feed components using inexpensive raw materials. Sustain Energ Fuels 1(4):923–931. https://doi.org/10.1039/C7SE00010C

    Article  CAS  Google Scholar 

  6. Johnson V, Singh M, Saini VS, Adhikari DK, Sista V, Yadav NK (1992) Bioemulsifier production by an oleaginous yeast Rhodotorula glutinis IIP 30. Biotechnol Lett 14(6):487–490. https://doi.org/10.1007/BF01023172

    Article  CAS  Google Scholar 

  7. Dubois M, Gilles K, Hamilton JK, Rebers PA, Smith F (1951) Colorimetric method for determination of sugars. Nature 168(4265). https://www.ncbi.nlm.nih.gov/pubmed/?term=SMITH%20F%5BAuthor%5D&cauthor=true&cauthor_uid=14875032. Accessed 4 Sept 2019

  8. Lowry OH, Rosebrough NJ, Al F, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193(1):265–275

    PubMed  CAS  Google Scholar 

  9. Satpute SK, Banpurkar AG, Dhakephalkar PK, Banat IM, Chopade BA (2010) Methods for investigating biosurfactants and bioemulsifiers: a review. Crit Rev Biotechnol 30(2):127–144. https://doi.org/10.3109/07388550903427280

    Article  PubMed  CAS  Google Scholar 

  10. Vilim V, Krajickova J (1991) Electrophoretic separation of large proteoglycans in large pore polyacrylamide gradient gels (1.32%-10% T) and a one step procedure for simultaneous staining of proteins and proteoglycans. Anal Biochem 197:34–39

    Article  CAS  Google Scholar 

  11. Castellane TCL, Campanharo JC, Coutinho LACID, Lopes ÉM, Lemos MVF, Lemos EGM (2017) Characterization of new exopolysaccharide production by Rhizobium tropici during growth on hydrocarbon substrate. Int J Biol Macromol 96:361–369. https://doi.org/10.1016/j.ijbiomac.2016.11.123

    Article  PubMed  CAS  Google Scholar 

  12. Violetta MR, Mazzoli R, Barello C, Fattori P, Giuffrida MG, Pessione E (2014) Combining LC-MS/MS, PMF and N-terminal amino acid sequencing for multiplexed characterization of a bacterial surfactant glycoprotein biosynthesized by Acinetobacter radioresistens S13. RSC Adv 4(21):10918–10927. https://doi.org/10.1039/C4RA00692E

    Article  CAS  Google Scholar 

  13. Cooper DG, Goldenberg BG (1987) Surface-active agents from two Bacillus species. Appl Environ Microbiol 53(2):224–229

    Article  CAS  Google Scholar 

  14. Uzoigwe C, Burgess JG, Ennis CJ, Rahman PKSM (2015) Bioemulsifiers are not biosurfactants and require different screening approaches. Front Microbiol 6:245. https://doi.org/10.3389/fmicb.2015.00245

    Article  PubMed  PubMed Central  Google Scholar 

  15. Fan Y, Tao W, Huang H, Li S (2017) Characterization of a novel bioemulsifier from Pseudomonas stutzeri. World J Microbiol Biotechnol 33:161. https://doi.org/10.1007/s11274-017-2326-2

    Article  PubMed  CAS  Google Scholar 

  16. Lukondeh T, Ashbolt NJ, Rogers PL (2003) Evaluation of Kluyveromyces marxianus FII 510700 grown on a lactose-based medium as a source of a natural bioemulsifier. J Ind Microbiol Biotechnol 30(12):715–720. https://doi.org/10.1007/s10295-003-0105-6

    Article  PubMed  CAS  Google Scholar 

  17. Amaral PFF, da Silva JM, Lehocky M, Barros-Timmons AMV, Coelho MAZ, Marrucho IM, Coutinho JAP (2006) Production and characterization of a bioemulsifier from Yarrowia lipolytica. Process Biochem 41(8):1894–1898. https://doi.org/10.1016/j.procbio.2006.03.029

    Article  CAS  Google Scholar 

  18. Cirigliano MC, Carman GM (1985) Purification and characterization of Liposan, a bioemulsifier from Candida lipolytica. Appl Environ Microbiol 50(4):846–850

    Article  CAS  Google Scholar 

  19. Sarubbo LA, Galba JMDL, Campos-Takaki MD (2006) Production and stability studies of the bioemulsifier obtained from a new strain of Candida glabarta UCP 1002. Electron J Biotechnol. https://doi.org/10.2225/vol9-issue4-fulltext-6

    Article  Google Scholar 

  20. Lorliam W, Akaracharanya A, Suzuki M, Ohkuma M, Tanasupawat S (2013) Diversity and fermentation products of xylose-utilizing yeasts isolated from buffalo feces in Thailand. Microbes Environ 28(3):354–360. https://doi.org/10.1264/jsme2.me13023

    Article  PubMed  PubMed Central  Google Scholar 

  21. Costa E, Teixidó N, Usall J, Atarés E, Viñas I (2002) The effect of nitrogen and carbon sources on growth of the biocontrol agent Pantoea agglomerans strain CPA-2. Lett Appl Microbiol 35:117–120

    Article  CAS  Google Scholar 

  22. Cooper DG, Paddock DA (1984) Production of a biosurfactant from Torulopsis bombicola. Appl Environ Microbiol 47(1):173–176

    Article  CAS  Google Scholar 

  23. Sarubbo LA, Marçal MC, Neves ML, Silva MP, Porto AL, Campos-Takaki GM (1997) Bioemulsifier production in batch culture using glucose as carbon source by Candida lipolytica. Appl Biochem Biotechnol 95(1):59–67

    Article  Google Scholar 

  24. Gargouri B, Contreras MD, Ammar S, Segura-Carretero A, Bouaziz M (2017) Biosurfactant production by the crude oil degrading Stenotrophomonas sp. B-2: chemical characterization, biological activities and environmental applications. Environ Sci Pollut Res 24(4):3769–3779. https://doi.org/10.1007/s11356-016-8064-4

    Article  CAS  Google Scholar 

  25. Otterstedt K, Larsson C, Bill RM, Ståhlberg A, Boles Eckhard, Hohman Sn, Gustafsson L (2004) EMBO Reports 5(5): 532–537. https://doi.org/10.1038/sj.embor.7400132

  26. Gudina EJ, Pereira JFB, Costa R, Evtuguin DV, Coutinho JAP, Teixeira JA, Rodrigues LR (2015) Novel bioemulsifier produced by a Paenibacillus strain isolated from crude oil. Microb Cell Fact. https://doi.org/10.1186/s12934-015-0197-5

    Article  PubMed  PubMed Central  Google Scholar 

  27. Ortega-de la Rosa ND, Vázquez-Vázquez JL, Huerta-Ochoa S, Gimeno M, Gutiérrez-Rojas M (2018) Stable bioemulsifiers are produced by Acinetobacter bouvetii UAM25 growing in different carbon sources. Bioproc Biosyst Eng 41:859–869. https://doi.org/10.1007/s00449-018-1920-5

    Article  CAS  Google Scholar 

  28. Khopade A, Biao R, Liu X, Mahadik K, Zhang L, Kokare C (2012) Production and stability studies of the biosurfactant isolated from marine Nocardiopsis sp. B4. Desalination 285:198–204. https://doi.org/10.1016/j.desal.2011.10.002

    Article  CAS  Google Scholar 

  29. van Gulik WM, Heijnen JJ (1995) A metabolic network stoichiometry analysis of microbial growth and product formation. Biotechnol Bioeng 48:681–698. https://doi.org/10.1002/bit.260480617

    Article  PubMed  Google Scholar 

  30. Patil JR, Chopade BA (2001) Studies on bioemulsifier production by Acinetobacter strains isolated from healthy human skin. J Appl Microbiol 91:290–298

    Article  CAS  Google Scholar 

  31. Velioglu Z, Urek RO (2015) Optimization of cultural conditions for biosurfactant production by Pleurotus djamor in solid state fermentation. J Biosci Bioeng 120(5):526–531. https://doi.org/10.1016/j.jbiosc.2015.03.007

    Article  PubMed  CAS  Google Scholar 

  32. Randhawa M (2019) Robbinsville , NJ,US. Topical composition containing glycerin and yeast extract (US Patent; Patent No- 10,206,870, B2, Date- Feb, 19, 2019)

  33. Kaplan N, Zosim Z, Rosenberg E (1987) Reconstitution of emulsifying activity of Acinetobacter calcoaceticus BD4 Emulsan by using pure polysaccharide and protein. Appl Environ Microbiol 53(2):440–446

    Article  CAS  Google Scholar 

  34. Zhao YH, Chen LY, Tian ZJ, Sun Y, Liu JB, Huang L (2016) Characterization and application of a novel bioemulsifier in crude oil degradation by Acinetobacter beijerinckii ZRS. J Basic Microbiol 56:184–195. https://doi.org/10.1002/jobm.201500487

    Article  PubMed  CAS  Google Scholar 

  35. Pines O, Bayer EA, Gutnick DL (1983) Localization of emulsan-like polymer associated with the cell surface of Acinetobacter calcoaceticus. J Bacteriol 154(2):893–905

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thankfully acknowledge Department of Biotechnology, Government of India (Project grant No.: BT/PR6909/PBD/26/391/2013, 21/03/2014) for financial assistance and Department of Biotechnology, IIT Kharagpur for all the research facilities. MB gratefully acknowledges Dr. Dineshkumar Ramalingam, Dr. Vivek Rangarajan and Dr. Ganeshan Subramaniam for their valuable technical inputs and suggestions throughout this study. CH gratefully acknowledges Department of Science and Technology (SEED Division), Government of India for financial assistance under the Scheme of Young Scientists and Technologists (SYST) (File no.: SP/YO/530/2018). RRK acknowledges Department of Biotechnology, Government of India (File no.: BT/PR17193/NER/95/474/2015, 13/02/2017).

Author information

Authors and Affiliations

  1. Department of Biotechnology, Indian Institute of Technology Kharagpur, West Bengal, Kharagpur, 721302, India

    Moumita Bhaumik, Gunaseelan Dhanarajan, Jayita Chopra, RaviRanjan Kumar, Chinmay Hazra & Ramkrishna Sen

Authors
  1. Moumita Bhaumik
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gunaseelan Dhanarajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jayita Chopra
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. RaviRanjan Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chinmay Hazra
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ramkrishna Sen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ramkrishna Sen.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

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

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 317 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bhaumik, M., Dhanarajan, G., Chopra, J. et al. Production, partial purification and characterization of a proteoglycan bioemulsifier from an oleaginous yeast. Bioprocess Biosyst Eng 43, 1747–1759 (2020). https://doi.org/10.1007/s00449-020-02361-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-020-02361-1

Keywords

Search

Navigation