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References

  1. Hawksworth DL, Kirk PM, Sutton BC, Pegler DN. 1996. Ainsworth & Bisby's dictionary of the fungi. Revista Do Instituto De Medicina Tropical De São Paulo 38: 17-19.
    CrossRef
  2. Kurtzman CP. 2011, pp. 233-234. The Yeasts, a Taxonomic Study, 5th Ed. Taylor & Francis, London.
  3. Wirth F, Goldani LZ. 2012. Epidemiology of Rhodotorula: an emerging pathogen. Interdiscip. Perspect. Infect. Dis. 2012: 465717.
    Pubmed PMC CrossRef
  4. Rose AH, Harrison JS. 1987. The Yeasts (ed by Rose AH, Harrison JS). 2: 181-250.
  5. Prabhala RH, Braune LM, Garewal HS, Watson RR. 2010. Influence of beta-carotene on immune functions. Ann. NY Acad. Sci. 691: 262-263.
    Pubmed CrossRef
  6. Hennekens CH. 1997. β-Carotene supplementation and cancer prevention. Nutrition 13: 697-699.
    CrossRef
  7. Andrew McWilliams. The global market for carotenoids, 2018. Available from https://www.bccresearch.com/market-research/food-and-beverage/the-global-market-for-carotenoids-fod025f.html. Accessed Jun. 2018.
  8. Buzzini P, Innocenti M, Turchetti B, Libkind D, Van BM, Mulinacci N. 2007. Carotenoid profiles of yeasts belonging to the genera Rhodotorula, Rhodosporidium, Sporobolomyces, and Sporidiobolus. Can. J. Microbiol. 53: 1024-1031.
    Pubmed CrossRef
  9. Villoutreix J. 1960. Les caroténoïdes de Rhodotorula mucilaginosa, étude de leur biosynthèse a l'aide de l'analyse de mutants et de l'emploi d'un inhibiteur de la caroténogénèse. Biochim. Biophys. Acta 40: 442-457.
    CrossRef
  10. Landolfo S, Ianiri G, Camiolo S, Porceddu A, Mulas G, Chessa R, et al. 2018. Microbiology 164: 78-87.
    Pubmed CrossRef
  11. Kot AM, Błażejak S, Gientka I, Kieliszek M, Bryś J. 2018. Torulene and torularhodin: "new" fungal carotenoids for industry? Microb. Cell. Fact. 17: 49.
    Pubmed PMC CrossRef
  12. Azmi Wamik TM, Kumari Priyanka. 2011. Production of a heat stable β-carotene with antioxidant activity by Rhodotorula sp. Int. Food Ferment. Technol. 1: 83-91.
  13. Perrier V, Dubreucq E, Galzy P. 1995. Fatty acid and carotenoid composition of Rhodotorula strains. Arch. Microbiol. 164: 173-179.
    Pubmed CrossRef
  14. Rodriguez-Concepcion M, Stange C. 2013. Biosynthesis of carotenoids in carrot: an underground story comes to light. Arch. Biochem. Biophys. 539: 110-116.
    Pubmed CrossRef
  15. Fromageot C, Tchang JL. 1938. Arch. Mikrobiol. 9: 424-433.
    CrossRef
  16. Bonner J, Sandoval A, Tang YW, Zechmeister L. 1946. Changes in polyene synthesis induced by mutation in a red yeast. Arch. Biochem. 10: 113.
  17. Razavi SH, Marc I. 2006. Effect of temperature and pH on the growth kinetics and carotenoid production by Sporobolomyces ruberrimus H110 using technical glycerol as carbon source. Iran. J. Chem. Chem. Eng. 25: 59-64.
  18. Ungureanu C, Ferdes M. 2012. Evaluation of Antioxidant and Antimicrobial Activities of Torularhodin. Adv. Sci. Lett. 18: 50-53(54).
    CrossRef
  19. Sakaki H, Nochide H, Komemushi S, Miki W. 2002. Effect of active oxygen species on the productivity of torularhodin by Rhodotorula glutinis No.21. J. Biosci. Bioeng. 93: 338-340.
    CrossRef
  20. Sakaki H, Nakanishi T, Tada A, Miki W, Komemushi S. 2001. Activation of torularhodin production by Rhodotorula glutinis using weak white light irradiation. J. Biosci. Bioeng. 92: 294-297.
    Pubmed CrossRef
  21. Du C, Li Y, Guo Y, Han M, Zhang W, Qian H. 2016. The suppression of torulene and torularhodin treatment on the growth of PC-3 xenograft prostate tumors. Biochem. Biophys. Res. Commun. 469: 1146-1152.
    Pubmed CrossRef
  22. Chao D, Guo Y, Cheng Y, Mei H, Zhang W, He Q. 2017. Torulene and torularhodin, protects human prostate stromal cells from hydrogen peroxide-induced oxidative stress damage through the regulation of Bcl-2/Bax mediated apoptosis. Free Radic. Res. 51: 113-123.
    Pubmed CrossRef
  23. Libkind D, Brizzio S, Van BM. 2004. Folia. Microbiol. 49: 19-25.
    Pubmed CrossRef
  24. Aksu Z, Eren AT. 2007. Biochem. Eng. J. 35: 107-113.
    CrossRef
  25. Tkáčová J FK, Klempová T, et al. 2015. Screening of carotenoid- producing Rhodotorula strains isolated from natural sources. Acta Chimica Slovaca 8: 34-38.
    CrossRef
  26. Bhosale P, Gadre RV. 2001. Production of β-carotene by a Rhodotorula glutinis mutant in sea water medium. Bioresour. Technol. 76: 53-55.
    CrossRef
  27. Wang SL, Sun JS, Han BZ, Wu XZ. 2010. Optimization of beta-carotene production by Rhodotorula glutinis using high hydrostatic pressure and response surface methodology. J. Food. Sci. 72: 325-329.
    Pubmed CrossRef
  28. Liu S, Li Q, Liu HL, Jia T, Xie DP. 2012. Mutation breeding of high-yield carotenoid producing Rhodotorula mucilaginosa by N~+ implantation and optimization of solid-state fermentation conditions for carotenoid production. Food Sci. 23: 244-248.
  29. Cong L, Chi Z, Li J, Wang X. 2007. J. Ocean. U. China. 6: 66-71.
    CrossRef
  30. Yolmeh M, Khomeiri M. 2016. Using physical and chemical mutagens for enhanced carotenoid production from Rhodotorula glutinis (PTCC 5256). Biocatal. Agric. Biotechnol. 8: 158-166.
    CrossRef
  31. Zhang Z, Zhang X, Tan T. 2014. Lipid and carotenoid production by Rhodotorula glutinis under irradiation/high- temperature and dark/low-temperature cultivation. Bioresour. Technol. 157: 149-153.
    Pubmed CrossRef
  32. Yen HW, Yang YC. 2012. Bioresour. Technol. 107: 539-541.
    Pubmed CrossRef
  33. Yen HW, Zhang Z. 2011. Bioresour. Technol. 102: 9279-9281.
    Pubmed CrossRef
  34. Hayman EP, Yokoyama H, Chichester CO, Simpson KL. 1974. J. Bacteriol. 120: 1339.
    Pubmed PMC CrossRef
  35. Simpson KL, Nakayama TO, Chichester CO. 1964. Biosynthesis of yeast carotenoids. J. Bacteriol. 88: 1688-1694.
    Pubmed PMC CrossRef
  36. Buzzini P, Martini A. 2000. Production of carotenoids by strains of Rhodotorula glutinis cultured in raw materials of agro-industrial origin. Bioresour. Technol. 71: 41-44.
    CrossRef
  37. Bhosale P, Gadre RV. 2010. Lett. Appl. Microbiol. 34: 349-353.
    Pubmed CrossRef
  38. Komemushi S, Sakaki H, Yokoyama H, Fujita T. 1994. Effect of barium and other metals on the growth of a D-lactic acid assimilating yeast Rhodotorula glutinis N21. J. Antibact. Antifungal. Agent 22: 583-587.
  39. Buzzini P, Martini A, Gaetani M, Turchetti B, Pagnoni UM, Davoli P. 2005. Optimization of carotenoid production by Rhodotorula graminis DBVPG 7021 as a function of trace element concentration by means of response surface analysis. Enzyme. Microb. Technol. 36: 687-692.
    CrossRef
  40. Kim BK, Park PK, Chae HJ, Kim EY. 2004. Korean. J. Chem. Eng. 21: 689-692.
    CrossRef
  41. Britton G, Singh RK, Malhotra HC, Goodwin TW, Ben-Aziz A. 1977. Biosynthesis of 1,2-dihydrocarotenoids in Rhodopseudo- monas viridis: experiments with inhibitors. Phytochemistry 16: 1561-1566.
    CrossRef
  42. Squina FM, Mercadante AZ. 2010. J. Food. Biochem. 29: 638-652.
    CrossRef
  43. Mata-Gómez LC, Montañez JC, Méndez-Zavala A, Aguilar CN. 2014. Biotechnological production of carotenoids by yeasts: an overview. Microb. Cell. Fact. 13: 12.
    Pubmed PMC CrossRef
  44. Fang TJ, Cheng Y-S. 1993. Improvement of Astaxanthin production by Phaffia rhodozyma through mutation and optimization of culture conditions. J. Ferment. Bioeng. 75: 466-469.
    CrossRef
  45. Frengova GI, Simova ED, Beshkova DM. 2004. Improvement of carotenoid-synthesizing yeast Rhodotorula rubra by chemical mutagenesis. Z. Naturforsch. C. 59: 99-103.
    Pubmed CrossRef
  46. Wang SL, Liu W, Wang HX, Lv CH. 2012. Ultra high- pressure and ion implantation combined mutagenesis to improve the production of β-carotene from red yeast. Adv. Mater. Res. II 554-556: 1165-1169.
    CrossRef
  47. Nasrabadi MRN, Razavi SH. 2011. Optimization of β-carotene production by a mutant of the lactose-positive yeast Rhodotorula acheniorum, from whey ultrafiltrate. Food. Sci. Biotechnol. 20: 445-454.
    CrossRef
  48. Sakaki H, Nakanishi T, Komemushi S, Namikawa K, Miki W. 2001. J. Clin. Biochem. Nutr. 30: 1-10.
    CrossRef
  49. Wang SL, Sha X, Wang HX. 2016. Improving yield of beta carotene in red yeast by using fermentation promoter. Food Nut. China. 22: 58-60.
  50. Marova I, Carnecka M, Halienova A, Certik M, Dvorakova T, Haronikova A. 2012. Use of several waste substrates for carotenoid-rich yeast biomass production. J. Environ. Manage 95: S338-S342.
    Pubmed CrossRef
  51. Husseiny SM, Abdelhafez AA, Ali AA, Sand HM, Husseiny SM, Abdelhafez AA, et al. 2017. Optimization of β-carotene production from Rhodotorula glutinis ATCC 4054 growing on agro-industrial substrate using plackett-burman design. P. Natl. A. Sci. India 3: 1-10.
    CrossRef
  52. Taskin M, Sisman T, Erdal S, Kurbanoglu EB. 2011. Use of waste chicken feathers as peptone for production of carotenoids in submerged culture of Rhodotorula glutinis MT-5. Eur. Food. Res. Technol. 233: 657-665.
    CrossRef
  53. Aksu Z, Eren AT. 2005. Carotenoids production by the yeast Rhodotorula mucilaginosa: use of agricultural wastes as a carbon source. Process. Biochem. 40: 2985-2991.
    CrossRef
  54. Petrik S, Marova I, Haronikova A, Kostovova I, Breierova E. 2013. Production of biomass, carotenoid and other lipid metabolites by several red yeast strains cultivated on waste glycerol from biofuel production - a comparative screening study. Ann. Microbiol. 63: 1537-1551.
    CrossRef
  55. Rodríguez-Sáiz MFJLDL, Barredo JL. 2010. Appl. Microbiol. Biotechnol. 88: 645-658.
    Pubmed CrossRef
  56. Zhu Z, Zhang S, Liu H, Shen H, Lin X, Yang F, et al. 2012. Nat. Commun. 3: 1112.
    Pubmed PMC CrossRef
  57. Sambles C, Middelhaufe S, Soanes D, Kolak D, Lux T, Moore K, et al. 2017. Genome sequence of the oleaginous yeast Rhodotorula toruloides strain CGMCC 2.1609. Genom. Data 13: 1-2.
    Pubmed PMC CrossRef
  58. Paul D, Magbanua Z, Arick M, French T, Bridges SM, Burgess SC, et al. 2014. Genome Sequence of the Oleaginous Yeast Rhodotorula glutinis ATCC 204091. Genome Announc. 2: 1-2.
    Pubmed PMC CrossRef
  59. Firrincieli A, Otillar R, Salamov A, Schmutz J, Khan Z, Redman RS, et al. 2015. Genome sequence of the plant growth promoting endophytic yeast Rhodotorula graminis WP1. Front. Microbiol. 6: 978.
    Pubmed PMC CrossRef
  60. Gan HM, Thomas BN, Cavanaugh NT, Morales GH, Mayers AN, Savka MA, et al. 2017. Whole genome sequencing of Rhodotorula mucilaginosa isolated from the chewing stick (Distemonanthus benthamianus): insights into Rhodotorula phy- logeny, mitogenome dynamics and carotenoid biosynthesis. PeerJ. 5: 1-18.
    Pubmed PMC CrossRef
  61. Deligios M, Fraumene C, Abbondio M. 2015. Draft genome sequence of Rhodotorula mucilaginosa, an emergent opportunistic pathogen. Genome Announc. 3: 1-2.
    Pubmed PMC CrossRef
  62. Tkavc R, Matrosova VY, Grichenko OE. 2017. Prospects for fungal bioremediation of acidic radioactive waste sites characterization and genome sequence of Rhodotorula taiwanensis MD1149. Front. Microbiol. 8: 2528.
    Pubmed PMC CrossRef
  63. Miccoli C, Palmieri D, Curtis FD, Lima G, Ianiri G, Castoria R. 2018. Complete genome sequence of the biocontrol agent yeast Rhodotorula kratochvilovae Strain LS11. Genome Announc. 6: 1-2.
    Pubmed PMC CrossRef
  64. Kim S, Kim J, Jung W, Kim J, Jung J. 2006. Biotechnol. Lett. 28: 897-904.
    Pubmed CrossRef
  65. Xu P, Bura R, Doty SL. 2015. Biotechnol. Bioeng. 63: 750-755.
    CrossRef
  66. Misawa N, Yamano S, Ikenaga H. 1991. Appl. Environ. Microbiol. 57: 1847-1849.
    Pubmed PMC CrossRef
  67. Kim JH, Kim SW, Nguyen DQA, Li H, Kim SB, Seo YG, et al. 2009. Production of β-carotene by recombinant Escherichia coli with engineered whole mevalonate pathway in batch and fed-batch cultures. Biotechnol. Bioprocess. Eng. 14: 559-564.
    CrossRef
  68. Yoon SH, Park HM, Kim JE, Lee SH, Choi MS, Kim JY, et al. 2010. Increased β-carotene production in recombinant Escherichia coli harboring an engineered isoprenoid precursor pathway with mevalonate addition. Biotechnol. Progr. 23: 599-605.
    Pubmed CrossRef
  69. Yoon SH, Lee SH, Das A, Ryu HK, Jang HJ, Kim JY, et al. 2009. J. Biotechnol. 140: 218-226.
    Pubmed CrossRef
  70. Yang J, Guo L. 2014. Biosynthesis of β-carotene in engineered E. coli using the MEP and MVA pathways. Microb. Cell Fact. 13: 160.
    Pubmed PMC CrossRef
  71. Zhao J, Li Q, Sun T, Zhu XN, Xu HT, Tang JL, et al. 2013. Engineering central metabolic modules of Escherichia coli for improving β-carotene production. Metab. Eng. 17: 42-50.
    Pubmed CrossRef
  72. Stephanopoulos G. 1999. Metabolic fluxes and metabolic engineering. Metab. Eng. 1: 1-11.
    Pubmed CrossRef
  73. Misawa N, Shimada H. 1998. Metabolic engineering for the production of carotenoids in non-carotenogenic bacteria and yeasts. J. Biotechnol. 59: 169-181.
    CrossRef
  74. Verwaal R, Wang J, Meijnen JP, Visser H, Sandmann G, Berg JAVD, et al. 2007. Appl. Environ. Microb. 73: 4342-4350.
    Pubmed PMC CrossRef
  75. Miura Y, Kondo K, Saito T, Shimada H, Fraser PD, Misawa N. 1998. Appl. Environ. Microbiol. 64: 1226-1229.
    Pubmed PMC CrossRef
  76. Shimada H, Kondo K, Fraser PD, Miura Y, Saito T, Misawa N. 1998. Increased carotenoid production by the food yeast Candida utilis through metabolic engineering of the isoprenoid pathway. Appl. Environ. Microbiol. 64: 2676-2680.
    Pubmed PMC CrossRef
  77. Araya-Garay JM, Feijoo-Siota L, Rosa-Dos-Santos F, Veiga- Crespo P, Villa TG. 2012. Construction of new Pichia pastoris X-33 strains for production of lycopene and β-carotene. Appl. Microbiol. Biotechnol. 93: 2483-2492.
    Pubmed CrossRef
  78. Abbott EP, Ianiri G, Castoria R, Idnurm A. 2013. Overcoming recalcitrant transformation and gene manipulation in Pucciniomycotina yeasts. Appl. Microbiol. Biotechnol. 97: 283-295.
    Pubmed CrossRef
  79. Liu Y, Koh CM, Sun L, Hlaing MM, Du M, Peng N, et al. 2013. Appl. Microbiol. Biotechnol. 97: 719-729.
    Pubmed CrossRef
  80. Pi HW, Anandharaj M, Kao YY, Lin YJ, Chang JJ, Lin WH. 2018. Engineering the oleaginous red yeast Rhodotorula glutinis for simultaneous β-carotene and cellulase production. Sci. Rep. 8: 10850.
    Pubmed PMC CrossRef

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Article

Review

J. Microbiol. Biotechnol. 2019; 29(4): 507-517

Published online April 28, 2019 https://doi.org/10.4014/jmb.1801.01022

Copyright © The Korean Society for Microbiology and Biotechnology.

Biosynthetic Pathway of Carotenoids in Rhodotorula and Strategies for Enhanced Their Production

Wei Tang 1, 2, Yue Wang 1, Jun Zhang 1, Yali Cai 1 and Zengguo He 1, 2*

1School of Medicine and Pharmacy, Ocean University of China, Qingdao 266000, P.R. China 2Marine Microbiological Engineering & Research Center, Marine Biomedical Research Institute of Qingdao, Qingdao 266000, P.R. China

Correspondence to:Zengguo  He
bioantai88@vip.163.com

Received: January 10, 2019; Accepted: March 9, 2019

Abstract

Rhodotorula is a group of pigment-producing yeasts well known for its intracellular biosynthesis of carotenoids such as β-carotene, γ-carotene, torulene and torularhodin. The great potential of carotenoids in applications in food and feed as well as in health products and cosmetics has generated a market value expected to reach over $2.0 billion by 2022. Due to growing public concern over food safety, the demand for natural carotenoids is rising, and this trend significantly encourages the use of microbial fermentation for natural carotenoid production. This review covers the biological properties of carotenoids and the most recent findings on the carotenoid biosynthetic pathway, as well as strategies for the metabolic engineering methods for the enhancement of carotenoid production by Rhodotorula. The practical approaches to improving carotenoid yields, which have been facilitated by advancements in strain work as well as the optimization of media and fermentation conditions, were summarized respectively

Keywords: Rhodotorula, carotenoids, biosynthetic pathway, strategies

Introduction

Rhodotorula is a genus of unicellular pigmented yeasts, part of the phylum Basidiomycota, family Cryptococcaceae, subfamily Rhodotorulodae [1]. According to the latest records of Mycobank (www.mycobank.org), there are 164 species in the genus Rhodotorula, including R. glutinis, R. toruloides, R. mucilaginosa, R. graminis and other varieties (Fig. 1). The cells of these Rhodotorula strains are polyphyletic in shape, appearing as subglobose, ovoid, ellipsoidal and elongated. The asexual reproduction of Rhodotorula is usually undertaken via multilateral and polar budding, whereas the sexual reproduction life cycle has occasionally occurred in the form of pseudohyphae in some strains [2].

Figure 1. Colony morphology of different strains of Rhodotorula. The data were collected from the CBS strain database. All the other strains were type strains except R. mucilaginosa var. mucilaginosa CBS 482.

Rhodotorula species are ubiquitous saprophytic yeasts and they can present in habitats with broad geographical varieties, e.g., from the equator to the poles and from land to the ocean. Strains in Rhodotorula are able to grow on different substrates in wide-ranging ecological conditions, such as air, soil, and manure, as well as in the bodies of animals, plants, and some lower organisms [3]. Rhodotorula is one of the dominant yeast flora in nutrient-poor aquatic environments, where they are actually reported to account for about 50% of the yeast population in seawater and fresh water [4].

One of the most notable characteristics of Rhodotorula is the formation of natural carotenoid biosynthesis. Enriched with pigments, Rhodotorula biomass alone can also serve as a high quality, single-cell protein source for utilization as a valuable feed additive.

Carotenoids have long been recognized worldwide as food additives and nutritional supplements, thanks to their valuable biological functionalities, such as antioxidative effects, immune response enhancement, preventive effects to cardiovascular disease, eye diseases and cancer, respectively [5, 6]. Nowadays, as a popular food additive, carotenoids are used broadly in the EU (listed as additive E160a), the US, Australia and New Zealand (listed as 160a) and other countries or regions, respectively. Statistically, the global market for carotenoids reached $1.5 billion in 2017 and should reach $2.0 billion by 2022, at a compound annual growth rate (CAGR) of 5.7% for the period of 2017-2022 [7]. Currently, more than 90% of commercial carotenoids are produced by chemical synthesis. Superior to chemical synthesis, the yeast-based production of carotenoids is natural and organic, entitled with a panel of extra advantages such as short production cycle time, environment-friendly usage, and ease of scale-up by manipulation through an engineered fermentation process. As one of the famous groups of carotenoid producers, strains of Rhodotorula may play important roles in the production of natural carotenoids in the future. In this study the biosynthetic pathway of carotenoids by Rhodotorula was reviewed and the strategies for carotenoid production were discussed.

Biosynthetic Pathway of Carotenoids in Rhodotorula

The biosynthetic pathway of carotenoids has been studied in a panel of Rhodotorula species. Buzzini et al. [8] indicated that R. minuta, R. glutinis, R. graminis, R. mucilaginosa and Rhodotorula sp. nov. possess analogical carotenoid profiles, with β-carotene, γ-carotene, torulene, and torularhodin representing the principal carotenoids in all these species. Villoutreix [9] also showed that a similar pigment profile was found in R. mucilaginosa. These approaches allowed the conclusion that the genus Rhodotorula possesses an identical or conserved carotenoid biosynthetic pathway (Fig. 2) [10, 11]. Carotenoid biosynthesis is known to follow the consecutive condensations of isoprenoid units into phytoene, the first colorless carotenoid in the pathway. The phytoene is continuously dehydrogenated and the conjugated double bond is extended until the formation of neurosporene, and subsequently, lycopene. There are two independent cyclization routes leading to two branches with either γ-carotene or torulene as the immediate products, which may further be converted to β-carotene and torularhodin respectively. Fig. 2 shows the most represent- ative steps and products in the carotenoid biosynthesis of Rhodotorula.

Figure 2. Biosynthetic pathway of carotenoids in Rhodotorula. crtI: phytoene desaturase encoding gene; crtYB: bifunctional lycopene cyclase/phytoene synthase encoding gene.

Principal Carotenoids and Their Physiology of Rhodotorula

β-Carotene and γ-Carotene

β-Carotene is one of the most well-known pigments having been widely used in medicines, health products, food additives, cosmetics, feed additives, and many products in other industries. It has also been approved as a food and feed additive for its dual functions in nutritive use and coloring in more than 50 countries and regions. Structurally, β-carotene is a fat-soluble, orange-yellow carotenoid with 11 conjugated double bonds and 2 retinyl groups (β-ionone ring). It is this large number of double bonds in its polyene chain and rings that makes it prone to be oxidized by free radicals. And this property endows it with antioxidant activity as well as enables its broad uses in food and feed applications [12]. For medical and or heath purpose use, β-carotene has been prescribed orally for the prevention of cancer, tumors and cardiovascular disease. Among the pigments in Rhodotorula species, β- carotene accounts for about 70% of total carotenoids [13].

γ-Carotene is the isomer of β-carotene, structurally containing 11 conjugated double bonds, 1 non-conjugated double bond and 1 retinyl group. Functionally, γ-carotene has vitamin A activity (though less than β-carotene) thanks to its single retinyl group [14]. It is formed via cyclization of lycopene through the enzymatic reaction facilitated by lycopene ε-cyclase.

Torulene and Torularhodin

As an acid pigment, torularhodin was first isolated from the genus Rhodotorula as early as the 1930s [15]. Subsequently, in 1946, Bonner et al. [16] announced the finding of torulene from R. rubra and they were able to overproduce torulene (amounting to 76% of the total carotenoids) by applying a mutant of R. rubra using quantitative chromatographic resolution. Interestingly it was not until the 1990s that torulene and torularhodin were regarded as potentially valuable substances [17]. Hence the number of works describing the properties of the two compounds increased quickly in the last decade.

Torulene is derived from the subtraction of 2H from γ-carotene, with the formation of an extra double bond in 13C (i.e., 13 double bonds), whereas torularhodin is formed from torulene by the substitution of one methyl group with one carboxyl group (that is, 14 double bonds). Since the two carotenoids contain the built-in β-ionone ring structure, the backbone of vitamin A, both of them can be the potential precursors of vitamin A. Compared to β-carotene, torulene and torularhodin have shown stronger antioxidant activity, thanks to the existence of the extra conjugated double bonds in 13C [18]. The extra double bond also endowed torularhodin with stronger capacity for scavenging hydrogen peroxide radicals as well as stronger resistance to the substrate degradation caused by singlet oxygen, compared to that of β-carotene, respectively [19]. It was also documented that high-torularhodin-production mutant can reduce the susceptibility to oxidative damage induced by active oxygen species [20].

In addition to antioxidant activity, in vivo anti-cancerous properties of the two carotenoids have also been demon- strated. In the anti-cancer supplementation experiments, compared to lycopene, both torulene and torularhodin performed much more significant inhibition of the growth of prostate cancer in nude mice by the induction of apoptosis of tumor cells [21]. Furtherly, it was confirmed that both of them facilitated protective activity on human prostate stromal cells from oxidative stress damage [22]. Meanwhile, the anti-microbial activity of the two carotenoids had also been confirmed, which revealed their potential usage on infection prevention particularly in implanted medical products and preparations that require natural antimicrobials (Kot et al., 2018) [11]. Lastly, torularhodin is one of the few carotenoids with carboxylic acid function [19].

Strategies for Enhanced Production of Carotenoids by Rhodotorula

Exploration of Native Carotenoid-Producing Strains

To satisfy the increasing demand for natural carotenoids, the development and utilization of high-yield carotenoid strains has become a research hotspot. So far Phaffia rhodozyma is the most popular red yeast used in the large- scale production of carotenoids, e.g., astaxanthin. Next to it, Rhodotorula may present as another red yeast that can be explored for production of carotenoids, particularly the production of mixtures of carotenoids at higher amounts. A wild strain of R. mucilaginosa CRUB 0138, an isolate from high-altitude Patagonian Lake Toncek, was found to produce carotenoids at a level of 234 ± 7 μg/g after 4 d incubation when the initial glucose concentration was adjusted to 1% [23]. Decent carotenoid yield at 33.2 mg/g was achieved by R. glutinis, a wild strain from refinery wastewater, when whey lactose-containing medium was applied [24]. A red yeast Rhodotorula sp. KF-104 of plant origin (isolated from vegetative parts of vine) was found to produce a mixture of carotenoids, including β-carotene, γ-carotene, torulene and torularhodin, respectively [25].

Improvement of Carotenoid-Synthesizing by Mutagenesis

Mutagenesis is a classical way for elevating the yield of carotenoids from wild strains, and the methods for mutation include UV radiation, N-methyl-N’-nitro-N-nitrosoguanidine (NTG), γ-irradiation, ethyl-methane sulfonate (EMS) and high hydrostatic pressure. Bhosale et al. [26] obtained a mutant 32 (2,048 μg/g) of a wild strain R. glutinis NCIM 3353 by UV radiation, which led to a yield increase of β-carotene in mutant 32 by 120-fold, as compared to that in the parent strain. Similarly, Wang et al. [27] was able to increase the yield of β-carotene by 57.89% in mutant strain RG6p, by using five repeated cycles of high hydrostatic pressure (HHP) mutation of the parent isolate R. glutinis GR6. Significant carotenoid yield improvement in R. mucilaginosa RM-1 was also documented by mutagenesis manipulation using N+ implantation of 10 keV and 2.0 × 1014 ion/cm2 [28]. Promising yield benefit (3-fold increase) was also achieved regarding carotenoid biosynthesis by the combined mutagenesis approaches, through a serial of UV, EMS and NTG mutagenesis [29]. In addition, the effects of different mutagenesis methods on the production of pigment from R. glutinis was evaluated, and it was found that mutagens could significantly increase the production of pigments, with UV irradiation as the better mutagenesis choice, compared to that of sodium azide (SA) [30].

Optimization Approaches on Cultivation Conditions for the Production of Carotenoids

The biosynthesis and accumulation of carotenoids are subjected to regulation by light, more specifically by photoinduction, which can improve carotenoid yields by promoting growth and cell density as well as by elevating the activity of enzymes involved in carotenoid biosynthesis. And this was well exemplified in R. glutinis by the work of Zhang et al. [31] that reported the irradiation caused significant biomass improvement and carotenoid production enhancement. In general, light-induced oxidative or radiation damage can limit the growth of microbes in some species, particularly those microorganisms that lack the proper intracellular photoprotective substances. Thanks to the endogenous carotenoid-empowered protection against light-induced damage, it has become possible for a panel of non-phototrophic yeasts and bacteria to grow and colonize in the natural world [32]. The work of Sakaki et al. [20] even reported the weak-white-light-facilitated yield improvement of carotenoids, in which a torularhodin increase of 180% was documented for R. glutinis. Simultaneously, the production of β-carotene in R. glutinis was also increased by 14%. Taken together, the author suggested that torularhodin may play a key role in the protection against oxidative damage caused by light irradiation. Practically, LED-derived irradiations of different colors could simply promote the growth of R. glutinis, among which irradiation by red LEDs induced the highest β-carotene production, followed in order by blue, green and white LEDs, respectively [33].

Temperature is another important factor affecting carotenoid biosynthesis. It can affect the propagation of carotenoid producers in general and influence the production of single carotenoid and carotenoid ratios along the biosynthesis pathway in specific. Hayman et al. [34] reported that the relative concentrations of individual carotenoids produced by R. glutinis were different when cultivated at 4°C and 25 °C, respectively. Similar results were observed in another strain of R. glutinis, 48-23T, in which the composition of carotenoids varied when different culture temperatures were applied. It was found that at 25°C the synthesized β-carotene, torulin and torulaihodin accounted for about 30% of the total carotenoids, respectively, whereas at 5°C, the proportion of β-carotene increased to 64%, by contrast the content of the other two carotenoids decreased significantly [35]. Buzzini and Martini [36] also reported that in R. glutinis the lower temperature (at 25°C) favored the synthesis of β-carotene and torulene whereas the higher temperature (30~35°C) is more suitable for the production of torularhodin. For R. glutinis it was even found that the optimal temperature of growth was different with that for carotenoid biosynthesis [37]. In this report the mutant 32 could grow well at optimum temperature of 30°C; however, the optimum temperature for the production of β-carotene was 20°C and its yield decreased with further reduction of the incubation temperature.

Many metal ions (Ba2+, Fe2+, Mg2+, Ca2+, Zn2+ and Co2+) were found capable to affect carotenogenesis in Rhodotorula, possibly because the activity of enzymes in the biosynthesis pathway were regulated, e.g., activated or inhibited, by those cations aforementioned, just as exemplified in desaturases in particular [38]. Buzzini et al. [39] reported that metal ions could selectively affect the carotenoid profile of R. graminis DBVPG 7021, and it was found in the research that Al3+ and Zn2+ promoted the production of β-carotene and γ-carotene, whereas Zn2+ and Mn2+ inhibited the production of torulene and torularhodin. In addition, the composing of exogenous solvents, chemicals or natural agents may affect the carotenoids in varied ways. Kim et al. [40] reported that the β-carotene content by R. glutinis increased by 35% when phenol was added to culture media at 500 ppm, and in contrast a decrease in torularhodin was observed when the phenol concentration was elevated. Diphenylamine, a widely used inhibitor, was also used to modify the carotenoid profile [41]. Diphenylamine was manipulated to block the sequential de-saturation reactions by inhibiting phytoene synthase, so that the accumulation of phytoene and other substances was enabled [41]. Squina and Mercadante [42] reported that the addition of 5 μmol of diphenylamine to the culture of R. rubra allowed more carotenoids to be produced in the broth, and interestingly in the experiment a significant reduction in torularhodin/ torulene ratio was identified in the cases for both R. rubra and R. glutinis. It is noteworthy that for both strains, the accumulation of β-carotene was enabled when further supplementation of diphenylamine (to a higher concentration at 10 μmol) was conducted.

As a group of secondary metabolites, most of the carotenoid biosynthesis initiated in the late logarithmic phase of yeasts near full growth was achieved, and their accumulation kept increasing during the stationary phase [43]. Nutrition wise, the producers can assimilate and metabolize diversified carbon sources, such as mono- saccharides, disaccharides and polysaccharides, organic acids and alcohols, and they can also rapidly utilize simple nitrogen sources (ammonium salt, nitrate, urea and amino acids) and complex mixtures (beef extract, yeast extract, malt extract, tryptone, etc.) [44]. For carotenoid production by Rhodotorula fermentation, the economical way through cost reduction is to use agro-industrial raw materials and by-products instead of using defined components as found in commercial media [36]. Table 1 summarized the examples regarding the production of carotenoids using low-cost raw materials. As shown, the production of carotenoids was affected by the choice of media components, such as the carbon and nitrogen sources, and the proportion of minerals and other components. Potential gains in yield can be attained simply by media optimization.

Table 1 . Carotenoids productivities under varied conditions by different strains of Rhodotorula.

StrategyStrainMethod/ScaleCarotenoidControlTreatmentReference
mg/gamg/Lamg/gamg/La
Mutagenesis
R. glutinis NCIM 3353UV radiationTotal carotenoids0.122.22.933[26]
R. rubra GED8NTGTotal carotenoids0.1872.670.648.12[46]
R. glutinis RG6High hydrostatic pressure(HHP)β-carotene-6.34-10.01[27]
R. glutinis NR-98Ultra high pressure (UHP)\ low energy nitrogen ionβ-carotene-6.03-17.36[46]
R. acheniorumUV, ethymethanesurfonate (EMS), and nitrosoguanidine (NTG)β-carotene2.3140.6010.69262.12[47]
Inducers
R. glutinisWhite lightβ-carotene3.6b-4.2b-[48]
Torulene29.3b-32.2b-
Torularhodin7.9b-14.2b-
R. glutinis RY-8Thiamineβ-carotene48.1250.152.9280.4[49]
Riboflavin51.8300.453.0323.3
Soybean oil51.4303.357.2366.1
Tomato juice50.6288.456.1342.2
Low-cost substrates
R. glutinis CCY20-2-26Wheyβ-carotene0.4817.931.0345.68[50]
R. glutinis ATCC 4054Rice branβ-carotene1.23c-3.20c-[51]
R. glutinis MT-5Waste chicken feathersTotal carotenoids5.76606.4792[52]
R. mucilaginosa CCY20-7-31Potato mediumβ-carotene0.164.311.8655.91[50]
R. mucilaginosa NRRL-2502Cotton seed oilTotal carotenoids-39.5-57.6[53]
R. aurantiacaWaste glycerolβ-carotene0.34-1.08-[54]

a Carotenoids content (mg/g cells dry weight or mg/L culture fluid); b Carotenoids content (mg/100 g cells dry weight); c Carotenoids content (mg/kg rice bran). .


Enhancing Carotenoid Production by Genetic Engineering and Metabolic Engineering Manipulations

Use of Genomic Tools to Characterize Carotenoid Biosynthesis in Rhodotorula

With the advancing of genetic engineering and metabolic engineering approaches, the construction of high-yield carotenoid strains as well as the maneuvering of the techniques for large-scale carotenoid production have become feasible. For Xanthophyllomyces dendrorhous (P. rhodozyma), a panel of genes encoding the key enzymes in the carotenoid pathway had been cloned and characterized with the astaxanthin biosynthesis pathway well illustrated [55]. However, so far there has been very limited progress for Rhodotorula spp. in this regard. This could be partially due to the limitation on available genomic data as well as the lack of functional annotation of the key genes, which impedes metabolic engineering manipulations aimed at the improvement of carotenoid production. Nevertheless, whole genome sequences of a few Rhodotorula spp. strains have been identified, and the establishing of the relevant bioinformatic data may benefit future research on aspects of the regulation of carotenoid biosynthesis, the pursuit of yield improvement, and the manipulation of relevant genes encoding other useful products, respectively (Table 2).

Table 2 . Comparative analysis on genomes data of strains of Rhodotorula.

OrganismAccession no.Size (Mb)GC%ScaffoldsGeneCharacteristicsReferences
R. toruloides NP11ALAU0000000020.2262948,171Triacylglycerol-producing[56]
R. toruloides CGMCC 2.1609LKER0000000033.3961.93659,820Inulinase activity[57]
R. glutinis ATCC 204091AEVR0000000020.4861.9293,359Lipids (>50% of its biomass) and antioxidants production[58]
R. graminis WP1JTAO0000000021.0367.76267,283Improves plant vigor, ferment both pentoses and hexoses, and degrade fermentation inhibitors[59]
R. mucilaginosa RIT389NIUW0000000019.6660.282507,065Isolated from the chewing stick (Distemonanthus benthamianus), and genomic regions containing the key genes for carotenoid production[60]
R. mucilaginosa C2.5t1JWTJ0000000019.9860.501,0346,413Isolated from a cacao seeds (Theobroma cacao L) in Cameroon, produce high carotenoid levels when grown in glycerol-containing media[61]
R. taiwanensis MD1149PJQD0000000019.5861.691817,122Resistant to acid (pH 2.3) and gamma radiation (66 Gy/h)[62]
R. kratochvilovae strain LS11PQDI0000000022.5666.6627,642Antagonistic activity against several plant pathogens[63]


The progress at the genetic level of Rhodotorula has been mostly focused on the identification of genes encoding the key enzymes and their distribution patterns along the carotenoid gene clusters. The whole genome of R. mucilaginosa RIT389 was sequenced with the identification of a genomic region associated with carotenoid, in which three genes encoding phytoene synthase (crtB), lycopene cyclase (crtY), and phytoene desaturase (crtI) were found closely located, whereas the gene encoding geranyl pyrophosphate synthase was located apart at a separate contig. The two other key genes, crtX and crtBY, that encoded carotenoid oxygenase and phytoene synthase/ lycopene cyclase, respectively, were found located in close proximity and convergently transcribed in all the species in Rhodotorula (except for R. mucilaginosa) [60]. On the base of the genome sequence of R. mucilaginosa C2.5t1, a set of genes involved in carotenogenesis were identified. Subsequent quantitative PCR showed that genes coding for 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (HMG1) and mevalonate kinase (ERG12) were induced at the end of the exponential growth phase. However, no clear trend of induction was observed for phytoene synthase/ lycopene cyclase (CAR2) and phytoene dehydrogenase (CAR1) encoding genes. It seems that the gene induction aforementioned was transient and occurred just at the beginning of carotenoid production, and somehow downstream-wise the expression level of CAR genes does not correlate with the amount of carotenoids produced [10].

Metabolic Engineering for Carotenoid Production in Non-Carotenogenic Bacteria and Yeasts

E. coli has been one of the most described microorganisms engineered to produce the exogenous carotenoids [64]. The cDNA of crtI, GGPP synthase (crtE) and crtYB genes from Erwinia uredovora were heterologously expressed in E. coli, showing lycopene accumulation in its transformants [65]. In addition, the crt genes derived from E. uredobora or E. herbicola were successfully used for the de novo biosynthesis of lycopene, β-carotene and zeaxanthin in E. coli [66]. In E. coli the innate apparatuses of the 2-C- methyl-D-erythritol 4-phosphate (MEP) pathway prepare isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), which serve as the precursors to enter the carotenoid biosynthesis pathway. IPP and DMAPP condense to form geranyl-pyrophosphate (GPP), which then synthesizes the precursor farnesyl-pyrophosphate (FPP) of β-carotene. The β-carotene can be synthesized by introducing genes crtE, crtB, crtI and crtY into E. coli. The expression of β-carotene in E. coli is shown in Table 3. The documented works were mainly focused on: (1) introducing β-carotene synthesis genes from different sources; (2) overexpressing synthase in the MEP pathway; (3) introducing mevalonate (MVA) pathway from eukaryotes into E. coli to increase IPP and FPP production; (4) reducing gene expression in competitive pathways; (5) adjusting ATP synthesis pathway, pentose phosphate pathway and TCA cycle to increase ATP and NADPH production.

Table 3 . Heterologous expression of β-carotene biosynthetic genes in E. coli.

StrategyMaximal yieldReferences
Engineered E. coli with a synthetic crt operon constructed to produce β-carotene390 mg/L[64]
Recombinant E. coli with engineered whole MVA pathways as well as harboring genes for β-carotene synthesis663 mg/L[67]
Recombinant E. coli harboring an engineered isoprenoid precursor pathway with mevalonate addition503 mg/L[68]
IPP and DMAPP supply can be increased significantly through the introduction of foreign MVA pathway into E. coli464 mg/L[69]
Improving the IPP and GPP concentration in the cell to increase β-carotene production, the optimized MEP pathway and hybrid MVA pathway have been introduced and co-expressed in an engineered E. coli strain3.2 g/L[70]
Combined engineering of MEP, β-carotene synthesis and central metabolic modules, a genetically stable E. coli strain was obtained which exhibited 74-fold yield increase over the wild type2.1 g/L[71]


Some yeasts, such as S. cerevisiae [72], and Candida utilis [73] have also been used to produce exogenous carotenoids through metabolic engineering manipulation. These yeasts are considered as food-grade organisms and have a mature genetic transformation system. They have been able to successfully produce carotenoids by insertion of caroteno- genic genes or metabolic pathways from E. uredovora, X. dendrorhous or Agrobacterium aurantiacum. Although wild-type S. cerevisiae cannot produce carotenoids, it does synthesize geranylgeranyl diphosphate (GGPP) which is an important precursor of carotenoid synthesis. In practice, two key genes from X. dendrorhous, crtYB and ctrI, were integrated into the genomic DNA of S. cerevisiae strain to complete the intact carotenoid biosynthesis pathway, and the manipulations ended up with successful expression of β-carotene [74]. Similarly, C. utilis does possess their potential precursors, but does not have the complete apparatuses for carotenoid biosynthesis. Through intro- duction of the three exogenous carotenoid genes, crtE, crtB, and crtI, a C. utilis strain producing 1.1 mg of pure lycopene per g (dry weight) of cells was obtained [75]. For the food yeast Candida utilis, manipulations of metabolic engineering were able bring about a seven-fold increase regarding lycopene production [76]. In addition, Pichia pastoris was also used to produce exogenous carotenoids through metabolic engineering manipulation. Araya-Garay et al. [77] designed and constructed two plasmids, pGAPZA- EBI* and pGAPZA-EBI*L*, which were integrated into P. pastoris genomic DNA, and the clones Pp-EBI and Pp- EBIL were used successfully for the production of lycopene or β-carotene, respectively.

It is noteworthy that the establishment of the afore- mentioned metabolic engineering processes of carotenoid biosynthesis are of great help in the manipulation toward the higher production of carotenoids by the engineered strains of choice. Actually, metabolic engineering techniques are becoming practical methods applied to Rhodotorula for the expression of exogenous carotenoids of interest as well as for the enhancement of the biosynthesis of endogenous carotenoid in specific. Abbott et al. [78] evaluated the feasibility of genetic engineering for different red yeasts by using common plasmids and transformation methods. The results showed that the success of transformation depends on the species and exogenous genes, which may be related to the G+C DNA content of several species. Consistent with this assumption, Liu et al. [79] reported that the codon optimization was the key to Agrobacterium tumefaciens- mediated transformation in R. toruloides. Pi et al. [80] transformed β-carotene biosynthesis genes (crtI, crtE, crtYB and tHMG1 from X. dendrorhous and Kluyveromyces marxianus) into R. glutinis genome. The transformant P4-10-9-63Y-14b produced β-carotene (27.13 ± 0.66 mg/g) 7-fold higher than the wild type. It is reasonable to expect that sophisticated metabolic engineering methods would allow the construction of suitable strains for the large-scale production of carotenoids of importance.

In conclusion, carotenoids not only serve as a class of excellent colorants, but also present as a group of natural products used for multi-dimensional applications. They have been widely used in food, medicine, health products cosmetics, and animal feed additives. The growing public concern on food safety urges the supply of carotenoids with natural origin, which prioritizes microbial carotenoid fermentation over other options, despite that the former not being the most cost-effective approach. Rhodotorula is well known for its potency in carotenoid production as well as for its feasibility to be used for carotenoid fermentation thanks to the short cycle time and the low cultivation cost. Systematic approaches have been made to enhance carotenoid production by Rhodotorula, utilizing advances in the strain work through natural breeding or mutagenesis, optimization of media and fermentation conditions, and metabolic engineering approaches on yield improvement as well as the attempts on the heterologous expression. It is reasonable to believe that the advent of elaborate strategies for mass carotenoid production by Rhodotorula are not far, given the availability of genetic and metabolic engineering potential and the maturity of advanced yeast-based industrial fermentation manufacturing systems.

Acknowledgments

This study was financially supported by the Postdoctoral Applied Research Project of Qingdao.

Conflict of Interest


The authors have no financial conflicts of interest to declare.

Fig 1.

Figure 1.Colony morphology of different strains of Rhodotorula. The data were collected from the CBS strain database. All the other strains were type strains except R. mucilaginosa var. mucilaginosa CBS 482.
Journal of Microbiology and Biotechnology 2019; 29: 507-517https://doi.org/10.4014/jmb.1801.01022

Fig 2.

Figure 2.Biosynthetic pathway of carotenoids in Rhodotorula. crtI: phytoene desaturase encoding gene; crtYB: bifunctional lycopene cyclase/phytoene synthase encoding gene.
Journal of Microbiology and Biotechnology 2019; 29: 507-517https://doi.org/10.4014/jmb.1801.01022

Table 1 . Carotenoids productivities under varied conditions by different strains of Rhodotorula.

StrategyStrainMethod/ScaleCarotenoidControlTreatmentReference
mg/gamg/Lamg/gamg/La
Mutagenesis
R. glutinis NCIM 3353UV radiationTotal carotenoids0.122.22.933[26]
R. rubra GED8NTGTotal carotenoids0.1872.670.648.12[46]
R. glutinis RG6High hydrostatic pressure(HHP)β-carotene-6.34-10.01[27]
R. glutinis NR-98Ultra high pressure (UHP)\ low energy nitrogen ionβ-carotene-6.03-17.36[46]
R. acheniorumUV, ethymethanesurfonate (EMS), and nitrosoguanidine (NTG)β-carotene2.3140.6010.69262.12[47]
Inducers
R. glutinisWhite lightβ-carotene3.6b-4.2b-[48]
Torulene29.3b-32.2b-
Torularhodin7.9b-14.2b-
R. glutinis RY-8Thiamineβ-carotene48.1250.152.9280.4[49]
Riboflavin51.8300.453.0323.3
Soybean oil51.4303.357.2366.1
Tomato juice50.6288.456.1342.2
Low-cost substrates
R. glutinis CCY20-2-26Wheyβ-carotene0.4817.931.0345.68[50]
R. glutinis ATCC 4054Rice branβ-carotene1.23c-3.20c-[51]
R. glutinis MT-5Waste chicken feathersTotal carotenoids5.76606.4792[52]
R. mucilaginosa CCY20-7-31Potato mediumβ-carotene0.164.311.8655.91[50]
R. mucilaginosa NRRL-2502Cotton seed oilTotal carotenoids-39.5-57.6[53]
R. aurantiacaWaste glycerolβ-carotene0.34-1.08-[54]

a Carotenoids content (mg/g cells dry weight or mg/L culture fluid); b Carotenoids content (mg/100 g cells dry weight); c Carotenoids content (mg/kg rice bran). .


Table 2 . Comparative analysis on genomes data of strains of Rhodotorula.

OrganismAccession no.Size (Mb)GC%ScaffoldsGeneCharacteristicsReferences
R. toruloides NP11ALAU0000000020.2262948,171Triacylglycerol-producing[56]
R. toruloides CGMCC 2.1609LKER0000000033.3961.93659,820Inulinase activity[57]
R. glutinis ATCC 204091AEVR0000000020.4861.9293,359Lipids (>50% of its biomass) and antioxidants production[58]
R. graminis WP1JTAO0000000021.0367.76267,283Improves plant vigor, ferment both pentoses and hexoses, and degrade fermentation inhibitors[59]
R. mucilaginosa RIT389NIUW0000000019.6660.282507,065Isolated from the chewing stick (Distemonanthus benthamianus), and genomic regions containing the key genes for carotenoid production[60]
R. mucilaginosa C2.5t1JWTJ0000000019.9860.501,0346,413Isolated from a cacao seeds (Theobroma cacao L) in Cameroon, produce high carotenoid levels when grown in glycerol-containing media[61]
R. taiwanensis MD1149PJQD0000000019.5861.691817,122Resistant to acid (pH 2.3) and gamma radiation (66 Gy/h)[62]
R. kratochvilovae strain LS11PQDI0000000022.5666.6627,642Antagonistic activity against several plant pathogens[63]

Table 3 . Heterologous expression of β-carotene biosynthetic genes in E. coli.

StrategyMaximal yieldReferences
Engineered E. coli with a synthetic crt operon constructed to produce β-carotene390 mg/L[64]
Recombinant E. coli with engineered whole MVA pathways as well as harboring genes for β-carotene synthesis663 mg/L[67]
Recombinant E. coli harboring an engineered isoprenoid precursor pathway with mevalonate addition503 mg/L[68]
IPP and DMAPP supply can be increased significantly through the introduction of foreign MVA pathway into E. coli464 mg/L[69]
Improving the IPP and GPP concentration in the cell to increase β-carotene production, the optimized MEP pathway and hybrid MVA pathway have been introduced and co-expressed in an engineered E. coli strain3.2 g/L[70]
Combined engineering of MEP, β-carotene synthesis and central metabolic modules, a genetically stable E. coli strain was obtained which exhibited 74-fold yield increase over the wild type2.1 g/L[71]

References

  1. Hawksworth DL, Kirk PM, Sutton BC, Pegler DN. 1996. Ainsworth & Bisby's dictionary of the fungi. Revista Do Instituto De Medicina Tropical De São Paulo 38: 17-19.
    CrossRef
  2. Kurtzman CP. 2011, pp. 233-234. The Yeasts, a Taxonomic Study, 5th Ed. Taylor & Francis, London.
  3. Wirth F, Goldani LZ. 2012. Epidemiology of Rhodotorula: an emerging pathogen. Interdiscip. Perspect. Infect. Dis. 2012: 465717.
    Pubmed KoreaMed CrossRef
  4. Rose AH, Harrison JS. 1987. The Yeasts (ed by Rose AH, Harrison JS). 2: 181-250.
  5. Prabhala RH, Braune LM, Garewal HS, Watson RR. 2010. Influence of beta-carotene on immune functions. Ann. NY Acad. Sci. 691: 262-263.
    Pubmed CrossRef
  6. Hennekens CH. 1997. β-Carotene supplementation and cancer prevention. Nutrition 13: 697-699.
    CrossRef
  7. Andrew McWilliams. The global market for carotenoids, 2018. Available from https://www.bccresearch.com/market-research/food-and-beverage/the-global-market-for-carotenoids-fod025f.html. Accessed Jun. 2018.
  8. Buzzini P, Innocenti M, Turchetti B, Libkind D, Van BM, Mulinacci N. 2007. Carotenoid profiles of yeasts belonging to the genera Rhodotorula, Rhodosporidium, Sporobolomyces, and Sporidiobolus. Can. J. Microbiol. 53: 1024-1031.
    Pubmed CrossRef
  9. Villoutreix J. 1960. Les caroténoïdes de Rhodotorula mucilaginosa, étude de leur biosynthèse a l'aide de l'analyse de mutants et de l'emploi d'un inhibiteur de la caroténogénèse. Biochim. Biophys. Acta 40: 442-457.
    CrossRef
  10. Landolfo S, Ianiri G, Camiolo S, Porceddu A, Mulas G, Chessa R, et al. 2018. Microbiology 164: 78-87.
    Pubmed CrossRef
  11. Kot AM, Błażejak S, Gientka I, Kieliszek M, Bryś J. 2018. Torulene and torularhodin: "new" fungal carotenoids for industry? Microb. Cell. Fact. 17: 49.
    Pubmed KoreaMed CrossRef
  12. Azmi Wamik TM, Kumari Priyanka. 2011. Production of a heat stable β-carotene with antioxidant activity by Rhodotorula sp. Int. Food Ferment. Technol. 1: 83-91.
  13. Perrier V, Dubreucq E, Galzy P. 1995. Fatty acid and carotenoid composition of Rhodotorula strains. Arch. Microbiol. 164: 173-179.
    Pubmed CrossRef
  14. Rodriguez-Concepcion M, Stange C. 2013. Biosynthesis of carotenoids in carrot: an underground story comes to light. Arch. Biochem. Biophys. 539: 110-116.
    Pubmed CrossRef
  15. Fromageot C, Tchang JL. 1938. Arch. Mikrobiol. 9: 424-433.
    CrossRef
  16. Bonner J, Sandoval A, Tang YW, Zechmeister L. 1946. Changes in polyene synthesis induced by mutation in a red yeast. Arch. Biochem. 10: 113.
  17. Razavi SH, Marc I. 2006. Effect of temperature and pH on the growth kinetics and carotenoid production by Sporobolomyces ruberrimus H110 using technical glycerol as carbon source. Iran. J. Chem. Chem. Eng. 25: 59-64.
  18. Ungureanu C, Ferdes M. 2012. Evaluation of Antioxidant and Antimicrobial Activities of Torularhodin. Adv. Sci. Lett. 18: 50-53(54).
    CrossRef
  19. Sakaki H, Nochide H, Komemushi S, Miki W. 2002. Effect of active oxygen species on the productivity of torularhodin by Rhodotorula glutinis No.21. J. Biosci. Bioeng. 93: 338-340.
    CrossRef
  20. Sakaki H, Nakanishi T, Tada A, Miki W, Komemushi S. 2001. Activation of torularhodin production by Rhodotorula glutinis using weak white light irradiation. J. Biosci. Bioeng. 92: 294-297.
    Pubmed CrossRef
  21. Du C, Li Y, Guo Y, Han M, Zhang W, Qian H. 2016. The suppression of torulene and torularhodin treatment on the growth of PC-3 xenograft prostate tumors. Biochem. Biophys. Res. Commun. 469: 1146-1152.
    Pubmed CrossRef
  22. Chao D, Guo Y, Cheng Y, Mei H, Zhang W, He Q. 2017. Torulene and torularhodin, protects human prostate stromal cells from hydrogen peroxide-induced oxidative stress damage through the regulation of Bcl-2/Bax mediated apoptosis. Free Radic. Res. 51: 113-123.
    Pubmed CrossRef
  23. Libkind D, Brizzio S, Van BM. 2004. Folia. Microbiol. 49: 19-25.
    Pubmed CrossRef
  24. Aksu Z, Eren AT. 2007. Biochem. Eng. J. 35: 107-113.
    CrossRef
  25. Tkáčová J FK, Klempová T, et al. 2015. Screening of carotenoid- producing Rhodotorula strains isolated from natural sources. Acta Chimica Slovaca 8: 34-38.
    CrossRef
  26. Bhosale P, Gadre RV. 2001. Production of β-carotene by a Rhodotorula glutinis mutant in sea water medium. Bioresour. Technol. 76: 53-55.
    CrossRef
  27. Wang SL, Sun JS, Han BZ, Wu XZ. 2010. Optimization of beta-carotene production by Rhodotorula glutinis using high hydrostatic pressure and response surface methodology. J. Food. Sci. 72: 325-329.
    Pubmed CrossRef
  28. Liu S, Li Q, Liu HL, Jia T, Xie DP. 2012. Mutation breeding of high-yield carotenoid producing Rhodotorula mucilaginosa by N~+ implantation and optimization of solid-state fermentation conditions for carotenoid production. Food Sci. 23: 244-248.
  29. Cong L, Chi Z, Li J, Wang X. 2007. J. Ocean. U. China. 6: 66-71.
    CrossRef
  30. Yolmeh M, Khomeiri M. 2016. Using physical and chemical mutagens for enhanced carotenoid production from Rhodotorula glutinis (PTCC 5256). Biocatal. Agric. Biotechnol. 8: 158-166.
    CrossRef
  31. Zhang Z, Zhang X, Tan T. 2014. Lipid and carotenoid production by Rhodotorula glutinis under irradiation/high- temperature and dark/low-temperature cultivation. Bioresour. Technol. 157: 149-153.
    Pubmed CrossRef
  32. Yen HW, Yang YC. 2012. Bioresour. Technol. 107: 539-541.
    Pubmed CrossRef
  33. Yen HW, Zhang Z. 2011. Bioresour. Technol. 102: 9279-9281.
    Pubmed CrossRef
  34. Hayman EP, Yokoyama H, Chichester CO, Simpson KL. 1974. J. Bacteriol. 120: 1339.
    Pubmed KoreaMed CrossRef
  35. Simpson KL, Nakayama TO, Chichester CO. 1964. Biosynthesis of yeast carotenoids. J. Bacteriol. 88: 1688-1694.
    Pubmed KoreaMed CrossRef
  36. Buzzini P, Martini A. 2000. Production of carotenoids by strains of Rhodotorula glutinis cultured in raw materials of agro-industrial origin. Bioresour. Technol. 71: 41-44.
    CrossRef
  37. Bhosale P, Gadre RV. 2010. Lett. Appl. Microbiol. 34: 349-353.
    Pubmed CrossRef
  38. Komemushi S, Sakaki H, Yokoyama H, Fujita T. 1994. Effect of barium and other metals on the growth of a D-lactic acid assimilating yeast Rhodotorula glutinis N21. J. Antibact. Antifungal. Agent 22: 583-587.
  39. Buzzini P, Martini A, Gaetani M, Turchetti B, Pagnoni UM, Davoli P. 2005. Optimization of carotenoid production by Rhodotorula graminis DBVPG 7021 as a function of trace element concentration by means of response surface analysis. Enzyme. Microb. Technol. 36: 687-692.
    CrossRef
  40. Kim BK, Park PK, Chae HJ, Kim EY. 2004. Korean. J. Chem. Eng. 21: 689-692.
    CrossRef
  41. Britton G, Singh RK, Malhotra HC, Goodwin TW, Ben-Aziz A. 1977. Biosynthesis of 1,2-dihydrocarotenoids in Rhodopseudo- monas viridis: experiments with inhibitors. Phytochemistry 16: 1561-1566.
    CrossRef
  42. Squina FM, Mercadante AZ. 2010. J. Food. Biochem. 29: 638-652.
    CrossRef
  43. Mata-Gómez LC, Montañez JC, Méndez-Zavala A, Aguilar CN. 2014. Biotechnological production of carotenoids by yeasts: an overview. Microb. Cell. Fact. 13: 12.
    Pubmed KoreaMed CrossRef
  44. Fang TJ, Cheng Y-S. 1993. Improvement of Astaxanthin production by Phaffia rhodozyma through mutation and optimization of culture conditions. J. Ferment. Bioeng. 75: 466-469.
    CrossRef
  45. Frengova GI, Simova ED, Beshkova DM. 2004. Improvement of carotenoid-synthesizing yeast Rhodotorula rubra by chemical mutagenesis. Z. Naturforsch. C. 59: 99-103.
    Pubmed CrossRef
  46. Wang SL, Liu W, Wang HX, Lv CH. 2012. Ultra high- pressure and ion implantation combined mutagenesis to improve the production of β-carotene from red yeast. Adv. Mater. Res. II 554-556: 1165-1169.
    CrossRef
  47. Nasrabadi MRN, Razavi SH. 2011. Optimization of β-carotene production by a mutant of the lactose-positive yeast Rhodotorula acheniorum, from whey ultrafiltrate. Food. Sci. Biotechnol. 20: 445-454.
    CrossRef
  48. Sakaki H, Nakanishi T, Komemushi S, Namikawa K, Miki W. 2001. J. Clin. Biochem. Nutr. 30: 1-10.
    CrossRef
  49. Wang SL, Sha X, Wang HX. 2016. Improving yield of beta carotene in red yeast by using fermentation promoter. Food Nut. China. 22: 58-60.
  50. Marova I, Carnecka M, Halienova A, Certik M, Dvorakova T, Haronikova A. 2012. Use of several waste substrates for carotenoid-rich yeast biomass production. J. Environ. Manage 95: S338-S342.
    Pubmed CrossRef
  51. Husseiny SM, Abdelhafez AA, Ali AA, Sand HM, Husseiny SM, Abdelhafez AA, et al. 2017. Optimization of β-carotene production from Rhodotorula glutinis ATCC 4054 growing on agro-industrial substrate using plackett-burman design. P. Natl. A. Sci. India 3: 1-10.
    CrossRef
  52. Taskin M, Sisman T, Erdal S, Kurbanoglu EB. 2011. Use of waste chicken feathers as peptone for production of carotenoids in submerged culture of Rhodotorula glutinis MT-5. Eur. Food. Res. Technol. 233: 657-665.
    CrossRef
  53. Aksu Z, Eren AT. 2005. Carotenoids production by the yeast Rhodotorula mucilaginosa: use of agricultural wastes as a carbon source. Process. Biochem. 40: 2985-2991.
    CrossRef
  54. Petrik S, Marova I, Haronikova A, Kostovova I, Breierova E. 2013. Production of biomass, carotenoid and other lipid metabolites by several red yeast strains cultivated on waste glycerol from biofuel production - a comparative screening study. Ann. Microbiol. 63: 1537-1551.
    CrossRef
  55. Rodríguez-Sáiz MFJLDL, Barredo JL. 2010. Appl. Microbiol. Biotechnol. 88: 645-658.
    Pubmed CrossRef
  56. Zhu Z, Zhang S, Liu H, Shen H, Lin X, Yang F, et al. 2012. Nat. Commun. 3: 1112.
    Pubmed KoreaMed CrossRef
  57. Sambles C, Middelhaufe S, Soanes D, Kolak D, Lux T, Moore K, et al. 2017. Genome sequence of the oleaginous yeast Rhodotorula toruloides strain CGMCC 2.1609. Genom. Data 13: 1-2.
    Pubmed KoreaMed CrossRef
  58. Paul D, Magbanua Z, Arick M, French T, Bridges SM, Burgess SC, et al. 2014. Genome Sequence of the Oleaginous Yeast Rhodotorula glutinis ATCC 204091. Genome Announc. 2: 1-2.
    Pubmed KoreaMed CrossRef
  59. Firrincieli A, Otillar R, Salamov A, Schmutz J, Khan Z, Redman RS, et al. 2015. Genome sequence of the plant growth promoting endophytic yeast Rhodotorula graminis WP1. Front. Microbiol. 6: 978.
    Pubmed KoreaMed CrossRef
  60. Gan HM, Thomas BN, Cavanaugh NT, Morales GH, Mayers AN, Savka MA, et al. 2017. Whole genome sequencing of Rhodotorula mucilaginosa isolated from the chewing stick (Distemonanthus benthamianus): insights into Rhodotorula phy- logeny, mitogenome dynamics and carotenoid biosynthesis. PeerJ. 5: 1-18.
    Pubmed KoreaMed CrossRef
  61. Deligios M, Fraumene C, Abbondio M. 2015. Draft genome sequence of Rhodotorula mucilaginosa, an emergent opportunistic pathogen. Genome Announc. 3: 1-2.
    Pubmed KoreaMed CrossRef
  62. Tkavc R, Matrosova VY, Grichenko OE. 2017. Prospects for fungal bioremediation of acidic radioactive waste sites characterization and genome sequence of Rhodotorula taiwanensis MD1149. Front. Microbiol. 8: 2528.
    Pubmed KoreaMed CrossRef
  63. Miccoli C, Palmieri D, Curtis FD, Lima G, Ianiri G, Castoria R. 2018. Complete genome sequence of the biocontrol agent yeast Rhodotorula kratochvilovae Strain LS11. Genome Announc. 6: 1-2.
    Pubmed KoreaMed CrossRef
  64. Kim S, Kim J, Jung W, Kim J, Jung J. 2006. Biotechnol. Lett. 28: 897-904.
    Pubmed CrossRef
  65. Xu P, Bura R, Doty SL. 2015. Biotechnol. Bioeng. 63: 750-755.
    CrossRef
  66. Misawa N, Yamano S, Ikenaga H. 1991. Appl. Environ. Microbiol. 57: 1847-1849.
    Pubmed KoreaMed CrossRef
  67. Kim JH, Kim SW, Nguyen DQA, Li H, Kim SB, Seo YG, et al. 2009. Production of β-carotene by recombinant Escherichia coli with engineered whole mevalonate pathway in batch and fed-batch cultures. Biotechnol. Bioprocess. Eng. 14: 559-564.
    CrossRef
  68. Yoon SH, Park HM, Kim JE, Lee SH, Choi MS, Kim JY, et al. 2010. Increased β-carotene production in recombinant Escherichia coli harboring an engineered isoprenoid precursor pathway with mevalonate addition. Biotechnol. Progr. 23: 599-605.
    Pubmed CrossRef
  69. Yoon SH, Lee SH, Das A, Ryu HK, Jang HJ, Kim JY, et al. 2009. J. Biotechnol. 140: 218-226.
    Pubmed CrossRef
  70. Yang J, Guo L. 2014. Biosynthesis of β-carotene in engineered E. coli using the MEP and MVA pathways. Microb. Cell Fact. 13: 160.
    Pubmed KoreaMed CrossRef
  71. Zhao J, Li Q, Sun T, Zhu XN, Xu HT, Tang JL, et al. 2013. Engineering central metabolic modules of Escherichia coli for improving β-carotene production. Metab. Eng. 17: 42-50.
    Pubmed CrossRef
  72. Stephanopoulos G. 1999. Metabolic fluxes and metabolic engineering. Metab. Eng. 1: 1-11.
    Pubmed CrossRef
  73. Misawa N, Shimada H. 1998. Metabolic engineering for the production of carotenoids in non-carotenogenic bacteria and yeasts. J. Biotechnol. 59: 169-181.
    CrossRef
  74. Verwaal R, Wang J, Meijnen JP, Visser H, Sandmann G, Berg JAVD, et al. 2007. Appl. Environ. Microb. 73: 4342-4350.
    Pubmed KoreaMed CrossRef
  75. Miura Y, Kondo K, Saito T, Shimada H, Fraser PD, Misawa N. 1998. Appl. Environ. Microbiol. 64: 1226-1229.
    Pubmed KoreaMed CrossRef
  76. Shimada H, Kondo K, Fraser PD, Miura Y, Saito T, Misawa N. 1998. Increased carotenoid production by the food yeast Candida utilis through metabolic engineering of the isoprenoid pathway. Appl. Environ. Microbiol. 64: 2676-2680.
    Pubmed KoreaMed CrossRef
  77. Araya-Garay JM, Feijoo-Siota L, Rosa-Dos-Santos F, Veiga- Crespo P, Villa TG. 2012. Construction of new Pichia pastoris X-33 strains for production of lycopene and β-carotene. Appl. Microbiol. Biotechnol. 93: 2483-2492.
    Pubmed CrossRef
  78. Abbott EP, Ianiri G, Castoria R, Idnurm A. 2013. Overcoming recalcitrant transformation and gene manipulation in Pucciniomycotina yeasts. Appl. Microbiol. Biotechnol. 97: 283-295.
    Pubmed CrossRef
  79. Liu Y, Koh CM, Sun L, Hlaing MM, Du M, Peng N, et al. 2013. Appl. Microbiol. Biotechnol. 97: 719-729.
    Pubmed CrossRef
  80. Pi HW, Anandharaj M, Kao YY, Lin YJ, Chang JJ, Lin WH. 2018. Engineering the oleaginous red yeast Rhodotorula glutinis for simultaneous β-carotene and cellulase production. Sci. Rep. 8: 10850.
    Pubmed KoreaMed CrossRef