Skip to main content
Log in

Production and optimization of carotenoid-enriched dried distiller’s grains with solubles by Phaffia rhodozyma and Sporobolomyces roseus fermentation of whole stillage

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Whole stillage—a co-product of grain-based ethanol—is used as an animal feed in the form of dried distiller’s grain with solubles (DDGS). Since animals cannot synthesize carotenoids and animal feed is generally poor in carotenoids, about 30–120 ppm of total carotenoids are added to animal feed to improve animal health, enhance meat color and quality, and increase vitamin A levels in milk and meat. The main objective of this study was to produce carotenoid (astaxanthin and β-carotene)-enriched DDGS by submerged fermentation of whole stillage. Mono- and mixed cultures of red yeasts, Phaffia rhodozyma (ATCC 24202) and Sporobolomyces roseus (ATCC 28988), were used to produce astaxanthin and β-carotene. Media optimization was carried out in shake flasks using response surface methodology (RSM). Macro ingredients, namely whole stillage, corn steep liquor and glycerol, were fitted to a second-degree polynomial in RSM. Under optimized conditions, astaxanthin and β-carotene yields in mixed culture and P. rhodozyma monoculture were 5 and 278, 97, and 275 μg/g, respectively, while S. roseus produced 278 μg/g of β-carotene. Since the carotenoid yields are almost twice the quantity used in animal feed, the carotenoid-enriched DDGS has potential application as “value-added animal feed or feed blends.”

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. An GH, Kim CH, Choi ES, Rhee SK (1996) Medium optimization for cultivation of carotenoid hyperproducing Phaffia rhodozyma mutant HT-5F01C. J Ferment Bioeng 82:515–518

    Article  CAS  Google Scholar 

  2. An GH, Song JY, Chang KS, Lee BD, Chae HS, Jang BG (2004) Pigmentation and delayed oxidation of broiler chickens by the red carotenoid, astaxanthin, from chemical synthesis and the yeast, Xanthophyllomyces dendrorhus. Asian Aust J Anim Sci 17:1–6

    Google Scholar 

  3. An GH, Song JY, Kwak WK, Lee BD, Song KB, Choi JE (2006) Improved astaxanthin availability due to drying and rupturing of the red yeast, Xanthophyllomyces dendrorhus. Food Sci Biotechnol 15:506–510

    CAS  Google Scholar 

  4. BCC Research (2005) The global market for carotenoids: updated edition. BCC Research, Wellesley

  5. Buzzini P, Innocenti M, Turchetti B, Libkind D, van Broock M, Mulinacci N (2007) Carotenoid profiles of yeasts belonging to the genera Rhodotorula, Rhodosporidium, Sporobolomyces, and Sporidiobolus. Can J Microbiol 53:1024–1031

    Article  CAS  PubMed  Google Scholar 

  6. Chew BP (1995) Antioxidant vitamins affect food animal immunity and health. J Nutr 125:1804S–1808S

    CAS  PubMed  Google Scholar 

  7. Das A, Yoon S-H, Lee S-H, Kim J-Y, Oh D-K, Kim S-W (2007) An update on microbial carotenoid production: application of recent metabolic engineering tools. Appl Microbiol Biotechnol 77:505–512

    Article  CAS  PubMed  Google Scholar 

  8. Cruz JM, Parajó JC (1998) Improved astaxanthin production by Xanthophyllomyces dendrorhous growing on enzymatic wood hydrolysates containing glucose and cellobiose. Food Chem 63:479–484

    Article  CAS  Google Scholar 

  9. Davoli P, Mierau V, Weber RWS (2004) Carotenoids and fatty acids in red yeast Sporobolomyces roseus and Rhodotorula glutinis. Appl Biochem Microbiol 40:392–397

    Article  CAS  Google Scholar 

  10. Davoli P, Weber RWS (2002) Carotenoid pigments from the red mirror yeast, Sporobolomyces roseus. Mycologist 16:102–108

    Google Scholar 

  11. US Grains Council (2007) DDGS user handbook: a guide to distiller’s dried grain with solubles. US Grains Council, Washington, DC

  12. Dong QL, Zhao XM (2004) In situ carbon dioxide fixation in the process of natural astaxanthin production by a mixed culture of Haematococcus pluvialis and Phaffia rhodozyma. Catal Today 98:537–544

    Article  CAS  Google Scholar 

  13. Echavarri-Erasun C, Johnson EA (2004) Stimulation of astaxanthin formation in the yeast Xanthophyllomyces dendrorhus by the fungus Epicoccum nigrum. FEMS Yeast Res 4:511–519

    Article  CAS  PubMed  Google Scholar 

  14. Eonseon J, Polle JEW, Lee HK, Hyun SM, Chang M (2003) Xanthophylls in microalgae: from biosynthesis to biotechnological mass production and application. J Microbiol Biotechnol 13:165–174

    Google Scholar 

  15. Fang TJ, Cheng Y (1993) Improvement of astaxanthin production by Phaffia rhodozyma through mutation and optimization of culture conditions. J Ferment Bioeng 75:466–469

    Article  CAS  Google Scholar 

  16. Frengova GI, Beshkova DM (2009) Carotenoids from Rhodotorula and Phaffia: yeasts of biotechnological importance. J Ind Microbiol Biotechnol 36:163–180

    Article  CAS  PubMed  Google Scholar 

  17. Hayek MG (2000) Process for enhancing immune response in animals using β-carotene as a dietary supplement. US Patent. US 6133323

  18. Hayman GT, Mannarelli BM, Leathers TD (1995) Production of carotenoids by Phaffia rhodozyma grown on media composed of corn wet-milling co-products. J Ind Microbiol 14:389–395

    Article  CAS  Google Scholar 

  19. Holden JM, Eldridge AL, Beecher GR, Buzzard IM, Bhagwat S, Davis CS, Douglas LW, Gebhardt S, Haytowitz D, Schakel S (1999) Carotenoid content of US foods: an update of the database. J Food Comp Anal 12:169–196

    Article  CAS  Google Scholar 

  20. Hurley WL, Doane RM (1989) Recent developments in the roles of vitamins minerals in reproduction. J Dairy Sci 72:784–804

    Article  CAS  PubMed  Google Scholar 

  21. Jacobson CK, Jolly SO, Sedmark JJ, Skatrud TJ, Wasilevski JM (2000) Astaxanthin over-producing strains of Phaffia rhodozyma. Method for their cultivation and their use in animal feeds. US Patent US 6015684

  22. Johnson EA (2003) Phaffia rhodozyma: colorful odyssey. Int Microbiol 6:169–174

    Article  CAS  PubMed  Google Scholar 

  23. Johnson EA, Lewis MJ (1979) Astaxanthin formation by the yeast Phaffia rhodozyma. J Gen Microbiol 115:173–183

    CAS  Google Scholar 

  24. Kusdiyantini E, Gaudin P, Goma G, Blanc PJ (1998) Growth kinetics and astaxanthin production of Phaffia rhodozyma on glycerol as a carbon source during batch fermentation. Biotechnol Lett 20:929–934

    Article  CAS  Google Scholar 

  25. Maldonade IR, Rodriguez-Amaya DB, Scamparini ARP (2008) Carotenoids of yeasts isolated from the Brazilian ecosystem. Food Chem 107:145–150

    Article  CAS  Google Scholar 

  26. Mantzouridou F, Naziri E, Tsimidou MZ (2008) Industrial glycerol as a supplementary carbon source in the production of β-carotene by Blakslea trispora. J Agric Food Chem 56:2668–2675

    Article  CAS  PubMed  Google Scholar 

  27. Margalith P (1993) Enhancement if carotenoid synthesis by fungal metabolites. Appl Microbiol Biotechnol 38:664–666

    Article  CAS  Google Scholar 

  28. Meyer PS, du Preez JC (1994) Astaxanthin production by a Phaffia rhodozyma mutant on grape juice. World J Microbiol Biotechnol 10:178–183

    Article  CAS  Google Scholar 

  29. Nghiem NP, Montanti J, Johnston D (2009) Production of astaxanthin from corn fiber as a value added co-product of fuel ethanol fermentation. Appl Biochem Biotechnol 154:227–237

    Article  CAS  Google Scholar 

  30. Ni H, Chen Q, Ruan H, Yang Y, Li L, Wu G, Hu Y, He G (2007) Studies on the optimization of nitrogen sources for astaxanthin production by Phaffia rhodozyma. J Zhejiang Univ 8:365–370

    Article  CAS  Google Scholar 

  31. Noziere P, Graulet B, Lucas A, Martin B, Grolier P, Doreau M (2006) Carotenoids for ruminants: from forage to diary products. Anim Feed Sci Technol 131:418–450

    Article  CAS  Google Scholar 

  32. Nys Y (2000) Dietary carotenoids and egg yolk coloration—a review. Arch Geflugelkd 64:45–54

    Google Scholar 

  33. Parajó JC, Santos V, Vázquez M (1997) Optimization of carotenoid production by Phaffia rhodozyma cells grown on xylose. Process Biochem 33:181–187

    Article  Google Scholar 

  34. Park PK, Cho DH, Kim EY, Chu KH (2005) Optimization of carotenoid production by Rhodotorula glutinis using statistical experimental design. World J Microbiol Biotechnol 21:429–434

    Article  CAS  Google Scholar 

  35. Ramírez J, Obledo N, Arellano M, Herrera E (2006) Astaxanthin production by Phaffia rhodozyma in a fed-batch culture using a low cost medium feeding. e-Gnosis 4:5

    Google Scholar 

  36. Ramírez J, Gutierrez H, Gshaedler A (2001) Optimization of astaxanthin production by Phaffia rhodozyma through factorial design and response surface methodology. J Biotechnol 88:259–268

    Google Scholar 

  37. Renewable Fuel Association (2009) Report for Jan 2009. http://www.ethanolrfa.org/industry/statistics

  38. Tinoi R, Pakariyatham N, Deming RL (2006) Utilization of mustard waste isolates for improved production of astaxanthin by Xanthophyllomyces dendrorhous. J Ind Microbiol Biotechnol 33:309–314

    Article  CAS  PubMed  Google Scholar 

  39. Valduga E, Valério A, Treichel H, Júnior AF, Luccio MD (2009) Optimization of the production of total carotenoids by Sporidiobolus salmonicolor (CBS 2636) using response surface technique. Food Bioprocess Technol 2:415–421

    Article  CAS  Google Scholar 

  40. Van den Berg H, Faulks R, Fernando Granado H, Hirschberg J, Olmedilla B, Sandmann G, Southon S, Stahl W (2000) The potential for the improvement of carotenoid levels in foods and the likely systemic effects. J Sci Food Agric 80:880–912

    Article  Google Scholar 

  41. Vázquez M, Martin AM (1997) Optimization of Phaffia rhodozyma continuous culture through response surface methodology. Biotech Bioengg 57:314–320

    Article  Google Scholar 

  42. Venugopal V (2009) Carotenoids. In: Venugopal V (ed) Marine products for healthcare: functional and bioactive nutraceutical compounds from the ocean. CRC Press, Boca Raton, pp 221–235

  43. Wang W, Yu L, Zhou P (2006) Effect of fungal elicitors in growth, total carotenoids and astaxanthin formation by Xanthophyllomyces dendrorhous. Bioresource Technol 97:26–31

    Article  CAS  Google Scholar 

  44. Yang A, Tume RK (1993) A comparison of beta carotene splitting activity isolated from intestinal mucosa of pasture grazed sheep, goat and cattle. Biochem Mol Biol Int 30:209–217

    CAS  PubMed  Google Scholar 

  45. Yurkov AM, Vustin MM, Tyaglov BV, Maksimova IA, Sineokiy SP (2008) Pigmented basidiomycetous yeasts are a promising source of carotenoids and ubiquinone Q-10. Microbiology 77:1–6

    CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to the Department of Grain Science and Industry, Kansas State University for funding this project. The authors are indebted to Dr. Yasuaki Hiromasa, BioCore facility, Department of Biochemistry, KSU, for MALDI/TOF mass spectroscopy analyses, Dr. Karthik Venkateshan, BioMaterial Lab, KSU for his assistance in lyophilization, Dr. Mark Anderson of Stat-Ease for assistance with RSM and mixture design, Dr. Laura Maurmann, NMR manager, Department of Chemistry, KSU for conducting NMR analyses, and Dr. Keerthi Mandyam for her critical review and valuable input into manuscript preparation. Abengoa Inc. (Colwich, KS, USA) provided the whole stillage. This article is assigned contribution no. 10-083-J from the Kansas Agricultural Experiment Station (Manhattan, KS 66506, USA).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Praveen V. Vadlani.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(DOC 103 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ananda, N., Vadlani, P.V. Production and optimization of carotenoid-enriched dried distiller’s grains with solubles by Phaffia rhodozyma and Sporobolomyces roseus fermentation of whole stillage. J Ind Microbiol Biotechnol 37, 1183–1192 (2010). https://doi.org/10.1007/s10295-010-0765-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-010-0765-y

Keywords

Navigation