Exploitation of genus Rhodosporidium for microbial lipid production

  • Jingyang XuEmail author
  • Dehua Liu


Oleaginous microorganisms are receiving significant attention worldwide for their utility in biodiesel production and the potentiality to produce some specialty-type lipids. There is an increasing interest in isolation/adaption of robust microbe strains and design of innovative fermentation processes to make microbial lipid production a more efficient and economically feasible bio-process. Currently, the genus Rhodosporidium has been considered an important candidate, for the reason that several strains belonging to this genus have shown excellent capabilities of lipid accumulation, broad adaptabilities to various substrates, and co-production of some carotenoids. This paper reviews the current trends in the exploitation of Rhodosporidium species for microbial lipid production, including the utilization of various (single or mixed, pure or waste-derived) substrates, progress of genetic modification and metabolic engineering, innovations in fermentation mode, lipid characterizations and their potential applications. Finally, the constraints and perspectives of cultivating Rhodosporidium species for lipid production are also discussed.


Feedstock Fermentation Metabolic engineering Microbial lipid Rhodosporidium 



The authors express their gratitude to the support from China Scholarship Council and Zhejiang Provincial Education Department Support Program (Y201533267).


  1. Abbott E, Ianiri G, Castoria R, Idnurm A (2013) Overcoming recalcitrant transformation and gene manipulation in Pucciniomycotina yeasts. Appl Microbiol Biotechnol 97:283–295CrossRefGoogle Scholar
  2. Bommareddy R, Sabra W, Maheshwari G, Zeng A (2015) Metabolic network analysis and experimental study of lipid production in Rhodosporidium toruloides grown on single and mixed substrates. Microb Cell Fact 14:36CrossRefGoogle Scholar
  3. Bommareddy R, Sabra W, Zeng A (2017) Glucose-mediated regulation of glycerol uptake in Rhodosporidium toruloides: Insights through transcriptomic analysis on dual substrate fermentation. Eng life Sci. doi: 10.1002/elsc.201600010 Google Scholar
  4. Chen Z, Liu P, Liu Y, Tang H, Chen Y, Zhang L (2014) Identification and characterization of a type-2 diacylglycerol acyltransferase (DGAT2) from Rhodosporidium diobovatum. Antonie Van Leeuwenhoek 106:1127–1137CrossRefGoogle Scholar
  5. Cui J, He S, Ji X, Lin L, Wei Y, Zhang Q (2016) Identification and characterization of a novel bifunctional Delta(12) Delta(15)-fatty acid desaturase gene from Rhodosporidium kratochvilovae. Biotechnol Lett 38:1155–1164CrossRefGoogle Scholar
  6. Davies R (1988) Yeast oil from cheese whey: process development. In: Moreton R (ed) Single cell oil. Longman Scientific & Technical, Harlow, pp 99–145Google Scholar
  7. Davies R, Holdsworth J (1992) Synthesis of lipids in yeasts: biochemistry, physiology and production. Adv Appl Lipid Res 1: 119–159Google Scholar
  8. Fei Q, O’Brien M, Nelson R, Chen X, Lowell A, Dowe N (2016) Enhanced lipid production by Rhodosporidium toruloides using different fed-batch feeding strategies with lignocellulosic hydrolysate as the sole carbon. Biotechnol Biofuels 9:130CrossRefGoogle Scholar
  9. Fillet S, Gibert J, Suarez B, Lara A, Ronchel C, Adrio J (2015) Fatty alcohols production by oleaginous yeast. J Ind Microbiol Biotechnol 42:1463–1472CrossRefGoogle Scholar
  10. Garay L, Sitepu I, Cajka T, Chandra I, Shi S, Lin T, German J, Fiehn O, Boundy-Mills K (2016) Eighteen new oleaginous yeast species. J Ind Microbiol Biotechnol 43:887–900CrossRefGoogle Scholar
  11. Gen Q, Wang Q, Chi Z (2014) Direct conversion of cassava starch into single cell oil by co-cultures of the oleaginous yeast. Renew Energy 62:522–526CrossRefGoogle Scholar
  12. Gierhart D (1984a) Multistage process for the preparation of fats and oils. US patent 4485172Google Scholar
  13. Gierhart D (1984b) Preparation of fats and oils. US patent 4485173Google Scholar
  14. Harde S, Wang Z, Horne M, Zhu J, Pan X (2016) Microbial lipid production from SPORL-pretreated Douglas fir by Mortierella isabellina. Fuel 175:64–74CrossRefGoogle Scholar
  15. Huang X, Liu J, Lu L, Peng K, Yang G, Liu J (2016) Culture strategies for lipid production using acetic acid as sole carbon source by Rhodosporidium toruloides. Bioresour Technol 206:141–149CrossRefGoogle Scholar
  16. Kiran E, Salakkam A, Trzcinski A, Bakir U, Webb C (2012) Enhancing the value of nitrogen from rapeseed meal for microbial oil production. Enzym Microb Technol 50:337–342CrossRefGoogle Scholar
  17. Kiran E, Trzcinski A, Webb C (2013) Microbial oil produced from biodiesel by-products could enhance overall production. Bioresour Technol 129:650–654CrossRefGoogle Scholar
  18. Koh C, Liu Y, Moehninsi, Du M, Ji L (2014) Molecular characterization of KU70 and KU80 homologues and exploitation of a KU70-deficient mutant for improving gene deletion frequency in Rhodosporidium toruloides. BMC Microbiol 14:50CrossRefGoogle Scholar
  19. Koutinas A, Chatzifragkou A, Kopsahelis N, Papanikolaou S, Kookos I (2014) Design and techno-economic evaluation of microbial oil production as a renewable resource for biodiesel and oleochemical production. Fuel 116:566–577CrossRefGoogle Scholar
  20. Kumar S, Kushwaha H, Bachhawat A, Raghava G, Ganesan K (2012) Genome Sequence of the Oleaginous Red Yeast Rhodosporidium toruloides MTCC 457. Eukaryot Cell 11:1083–1084CrossRefGoogle Scholar
  21. Lee J, Chen L, Shi J, Trzcinski A, Chen W (2014) Metabolomic profiling of Rhodosporidium toruloides grown on glycerol for carotenoid production during different growth phases. J Agric Food Chem 62:10203–10209CrossRefGoogle Scholar
  22. Lee J, Chen L, Cao B, Chen W (2016) Engineering Rhodosporidium toruloides with a membrane transporter facilitates production and separation of carotenoids and lipids in a bi-phasic culture. Appl Microbiol Biotechnol 100:869–877CrossRefGoogle Scholar
  23. Leiva-Candia D, Tsakona S, Kopsahelis N, Garcia I, Papanikolaou S, Dorado M, Koutinas A (2015) Biorefining of by-product streams from sunflower-based biodiesel production plants for integrated synthesis of microbial oil and value-added co-products. Bioresouc Technol 190: 57–65CrossRefGoogle Scholar
  24. Li Y, Zhao Z, Bai F (2007) High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture. Enzym Microb Technol 41:312–317CrossRefGoogle Scholar
  25. Liang M, Jiang J (2013) Advancing oleaginous microorganisms to produce lipid via metabolic engineering technology. Prog Lipid Res 52:395–408CrossRefGoogle Scholar
  26. Lin X, Wang Y, Zhang S, Zhu Z, Zhou Y, Sun W, Wang X, Zhao Z (2014a) Functional integration of multiple genes into the genome of the oleaginous yeast Rhodosporidium toruloides. FEMS Yeast Res 14:547–555CrossRefGoogle Scholar
  27. Lin J, Li S, Sun M, Zhang C, Yang W, Zhang Z, Li X, Li S (2014b) Microbial lipid production by oleaginous yeast in D-xylose solution using a two-stage culture mode. RSC Adv 4:34944–34949CrossRefGoogle Scholar
  28. Ling J, Nip S, Shim H (2013) Enhancement of lipid productivity of Rhodosporidium toruloides in distillery wastewater by increasing cell density. Bioresour Technol 146:301–309CrossRefGoogle Scholar
  29. Ling J, Nip S, Cheok W, de Toledo R, Shim H (2014) Lipid production by a mixed culture of oleaginous yeast and microalga from distillery and domestic mixed wastewater. Bioresour Technol 173:132–139CrossRefGoogle Scholar
  30. Liu Y, Koh C, Sun L, Hlaing M, Du M, Peng N, Ji L (2013) Characterization of glyceraldehyde-3-phosphate dehydrogenase gene RtGPD1 and development of genetic transformation method by dominant selection in oleaginous yeast Rhodosporidium toruloides. Appl Microbiol Biotechnol 97:719–729CrossRefGoogle Scholar
  31. Liu Y, Koh C, Ngoh S, Ji L (2015) Engineering an efficient and tight d-amino acid-inducible gene expression system in Rhodosporidium Rhodotorula species. Microb Cell Fact 14:170CrossRefGoogle Scholar
  32. Liu Y, Zhang M, Wang T, Shi X, Li J, Jia L, Tang H, Zhang L (2016) Two acetyl-CoA synthetase isoenzymes are encoded by distinct genes in marine yeast Rhodosporidium diobovatum. Biotechnol Lett 38:417–423CrossRefGoogle Scholar
  33. Luque L, Orr V, Chen S, Westerhof R, Oudenhoven S, Rossum G, Kersten S, Berruti F, Rehmann L (2016) Lipid accumulation from pinewood pyrolysates by Rhodosporidium diobovatum and Chlorella vulgaris for biodiesel production. Bioresour Technol 214:660–669CrossRefGoogle Scholar
  34. Mannazzu I, Landolfo S, Silva T, Buzzini P (2015) Red yeasts and carotenoid production: outlining a future for non-conventional yeasts of biotechnological interest. World J Microbiol Biotechnol 31:1665–1673CrossRefGoogle Scholar
  35. Matsakas L, Bonturi N, Miranda E, Rova U, Christakopoulos P (2015) High concentrations of dried sorghum stalks as a biomass feedstock for single cell oil production by Rhodosporidium toruloides. Biotechnol Biofuels 8:6CrossRefGoogle Scholar
  36. Moreton R (1985) Modification of fatty acid composition of lipid accumulating yeasts with cyclopropene fatty acid desaturase inhibitors. Appl Microbiol Biotechnol 22:41–45CrossRefGoogle Scholar
  37. Moreton R, Clode D (1988) Microbial desaturase enzyme inhibitors and their use in a method of producing lipids. US patent 4778630Google Scholar
  38. Morikawa Y, Zhao X, Liu D (2014) Biological co-production of ethanol and biodiesel from wheat straw- a case of dilute acid pretreatment. RSC Adv 4:37878–37888CrossRefGoogle Scholar
  39. Munch G, Sestric R, Sparling R, Levin D, Cicek N (2015) Lipid production in the under-characterized oleaginous yeasts Rhodosporidium babjevae and Rhodosporidium diobovatum from biodiesel-derived waste glycerol. Bioresour Technol 185:49–55CrossRefGoogle Scholar
  40. Papanikolaou S, Aggelis G (2010) Yarrowia lipolytica: A model microorganism used for the production of tailor-made lipids. Eur J Lipid Sci Technol 112:639–654CrossRefGoogle Scholar
  41. Papanikolaou S, Aggelis G (2011a) Lipids of oleaginous yeasts. Part Ι: Biochemistry of single cell oil production. Eur J Lipid Sci Technol 113:1031–1051CrossRefGoogle Scholar
  42. Papanikolaou S, Aggelis G (2011b) Lipids of oleaginous yeasts. Part ΙΙ: technology and potential applications. Eur J Lipid Sci Technol 113:1052–1073CrossRefGoogle Scholar
  43. Parreira T, Freitas C, Reis A, Roseiro J, Silva T (2015) Carbon concentration and oxygen availability affect lipid and carotenoid production by carob pulp syrup. Eng. Life Sci 15:815–823CrossRefGoogle Scholar
  44. Patel A, Pravez M, Deeba F, Pruthi V, Singh R, Pruthi P (2014) Boosting accumulation of neutral lipids in Rhodosporidium kratochvilovae HIMPA1 grown on hemp seed. Bioresour Technol 165:214–222CrossRefGoogle Scholar
  45. Patel A, Pruthi V, Singh R, Pruthi P (2015a) Synergistic effect of fermentable and non-fermentable carbon sources enhances TAG accumulation in oleaginous yeast Rhodosporidium kratochvilovae HIMPA1. Bioresour Technol 188:136–144CrossRefGoogle Scholar
  46. Patel A, Sindhu D, Arora N, Singh R, Pruthi V, Pruthi P (2015b) Biodiesel production from non-edible lignocellulosic biomass of Cassia fistula L. fruit pulp using oleaginous yeast Rhodosporidium kratochvilovae HIMPA1. Bioresour Technol 197:91–98CrossRefGoogle Scholar
  47. Polburee P, Yongmanitchai W, Honda K, Ohashi T, Yoshida T, Fujiyama K, Limtong S (2016) Lipid production from biodiesel-derived crude glycerol by Rhodosporidium fluviale DMKU-RK253 using temperature shift with high cell density. Biochem Eng J 112:208–218CrossRefGoogle Scholar
  48. Qi F, Kitahara Y, Wang Z, Zhao X, Du W, Liu D (2014) Novel mutant strains of Rhodosporidium toruloides by plasma mutagenesis approach and their tolerance for inhibitors in lignocellulosic hydrolysate. J Chem Technol Biotechnol 89:735–742CrossRefGoogle Scholar
  49. Qi F, Zhao X, Kitahara Y, Li T, Ou X, Du W, Liu D, Huang J (2017) Integrative transcriptomic and proteomic analysis of the mutant lignocellulosic hydrolysate-tolerant Rhodosporidium toruloides. Eng. Life Sci. doi: 10.1002/elsc.201500143 Google Scholar
  50. Shi J, Feng H, Lee J, Chen W (2013) Comparative proteomics profile of lipid-cumulating oleaginous yeast: An iTRAQ-coupled 2-D LC-MS MS analysis. PloS ONE 8:e85532CrossRefGoogle Scholar
  51. Signori L, Ami D, Posteri R, Giuzzi A, Mereghetti P, Porro D, Branduardi P (2016) Assessing an effective feeding strategy to optimize crude glycerol utilization as sustainable carbon source for lipid accumulation in oleaginous yeasts. Microb Cell Fact 15:75CrossRefGoogle Scholar
  52. Tai M, Stephanopoulos G (2013) Engineering the push and pull of lipid biosynthesis in oleaginous yeast Yarrowia lipolytica for biofuel production. Metab Eng 15:1–9CrossRefGoogle Scholar
  53. Tchakouteu S, Kalantzi O, Gardeli C, Koutinas A, Aggelis G, Papanikolaou S (2015) Lipid production by yeasts growing on biodiesel-derived crude glycerol strain selection and impact of substrate concentration on the fermentation efficiency. J Appl Microbiol 118:911–927CrossRefGoogle Scholar
  54. Tchakouteu S, Kopsahelis N, Chatzifragkou A, Kalantzi O, Stoforos N, Koutinas A, Aggelis G, Papanikolaou S (2017) Rhodosporidium toruloides cultivated in NaCl-enriched glucose-based media: Adaptation dynamics and lipid production. Eng. Life Sci. doi: 10.1002/elsc.201500125 Google Scholar
  55. Tully M, Gilbert H (1985) Transformation of Rhodosporidium toruloides. Gene 36:235–240CrossRefGoogle Scholar
  56. Vieira J, Ienczak J, Rossell C, Pradella J, Franco T (2014) Microbial lipid production- screening with yeasts grown on Brazilian molasses. Biotechnol Lett 36:2433–2442CrossRefGoogle Scholar
  57. Wang Z, Fu W, Xu H, Chi Z (2014) Direct conversion of inulin into cell lipid by an inulinase-producing yeast Rhodosporidium toruloides 2F5. Bioresour Technol 161:131–136CrossRefGoogle Scholar
  58. Wang Y, Lin X, Zhang S, Sun W, Ma S, Zhao Z (2016) Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides. Yeast 33:99–106CrossRefGoogle Scholar
  59. Wasylenko T, Ahn W, Stephanopoulos G (2015) The oxidative pentose phosphate pathway is the primary source of NADPH for lipid overproduction from glucose in Yarrowia lipolytica. Metab Eng 30:27–39CrossRefGoogle Scholar
  60. Wiebe M, Koivuranta K, Renttila M, Ruohonen L (2012) Lipid production in batch and fed-batch cultures of Rhodosporidium toruloides from 5 and 6 carbon carbohydrates. BMC Biotechnol 12:26CrossRefGoogle Scholar
  61. Wu S, Zhao X, Shen H, Wang Q, Zhao Z (2011) Microbial lipid production by Rhodosporidium toruloides under sulfate-limited conditions. Bioresour Technol 102:1803–1807CrossRefGoogle Scholar
  62. Xu J, Zhao X, Wang W, Du W, Liu D (2012) Microbial conversion of biodiesel byproduct glycerol to triacylglycerols by oleaginous yeast Rhodosporidium toruloides and the individual effect of some impurities on lipid production. Biochem Eng J 65:30–36CrossRefGoogle Scholar
  63. Xu J, Du W, Zhao X, Zhang G, Liu D (2013) Microbial oil production from various carbon sources and its use for biodiesel preparation. Biofuels Bioprod Biorefin 7:65–77CrossRefGoogle Scholar
  64. Xu P, Qiao K, Ahn W, Stephanopoulos G (2016) Engineering Yarrowia lipolytica as a platform for synthesis of drop-in transportation fuels and oleochemicals. Proc Natl Acad Sci 113:10848–10853CrossRefGoogle Scholar
  65. Xu J, Zhao X, Du W, Liu D (2017) Bioconversion of glycerol into lipids by Rhodosporidium toruloides in a two-stage process and characterization of lipid properties. Eng. Life Sci. doi: 10.1002/elsc.201600062 Google Scholar
  66. Yang F, Zhang S, Zhou Y, Zhu Z, Lin X, Zhao Z (2012) Characterization of the mitochondrial NAD(+)-dependent isocitrate dehydrogenase of the oleaginous yeast Rhodosporidium toruloides. Appl Microbiol Biotechnol 94:1095–1105CrossRefGoogle Scholar
  67. Yang X, Jin G, Gong Z, Shen H, Bai F, Zhao Z (2015) Recycling microbial lipid production wastes to cultivate oleaginous yeasts. Bioresour Technol 175:91–96CrossRefGoogle Scholar
  68. Zhang S, Ito M, Skerker J, Arkin A, Rao C (2016a) Metabolic engineering of the oleaginous yeast Rhodosporidium toruloides IFO0880 for lipid overproduction during high-density fermentation. Appl Microbiol Biotechnol 100:9393–9405CrossRefGoogle Scholar
  69. Zhang S, Skerker J, Rutter C, Maurer M, Arkin A, Rao C (2016b) Engineering Rhodosporidium toruloides for increased lipid production. Biotechnol Bioeng 113:1056–1066CrossRefGoogle Scholar
  70. Zhang X, Shen H, Yang X, Wang Q, Yu X, Zhao Z (2016c) Microbial lipid production by oleaginous yeasts on Laminaria residue hydrolysates. RSC Adv 6:26752–26756CrossRefGoogle Scholar
  71. Zhao X, Peng F, Du W, Liu C, Liu D (2012) Effects of some inhibitors on the growth and lipid accumulation of oleaginous yeast Rhodosporidium toruloides. Bioprocess Biosyst Eng 35:993–1004CrossRefGoogle Scholar
  72. Zhou W, Wang W, Li Y, Zhang Y (2013) Lipid production by Rhodosporidium toruloides Y2 in bioethanol wastewater and evaluation of biomass energetic yield. Bioresour Technol 127:435–440CrossRefGoogle Scholar
  73. Zhu Z, Zhang S, Liu H, Shen H, Lin X, Yang F, Zhou Y, Jin G, Ye M, Zou H, Zhao Z (2012) A multi-omic map of the lipid-producing yeast Rhodosporidium toruloides. Nat Commun 3:1112CrossRefGoogle Scholar
  74. Zhu Z, Ding Y, Gong Z, Yang L, Zhang S, Zhang C, Lin X, Shen H, Zou H, Xie Z, Yang F, Zhao X, Liu P, Zhao Z (2015) Dynamics of the Lipid Droplet Proteome of the Oleaginous Yeast Rhodosporidium toruloides. Eukaryot Cell 14:252–264CrossRefGoogle Scholar

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© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Zhejiang Police CollegeHangzhouPeople’s Republic of China
  2. 2.Department of Chemical EngineeringMassachusetts Institute of TechnologyCambridgeUSA
  3. 3.Department of Chemical Engineering, Institute of Applied ChemistryTsinghua UniversityBeijingPeople’s Republic of China

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