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The isolation and characterization of Pseudozyma sp. JCC 207, a novel producer of squalene

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Abstract

In examining the production of valuable compounds by marine microorganisms, we isolated a novel yeast strain that produces a large amount of squalene and several polyunsaturated fatty acids. Molecular and phylogenetic analyses of the ribosomal DNA suggest that the isolate belongs to the genus Pseudozyma, which comprises ustilaginomycetous anamorphic yeasts. The nucleotide sequence of an internally transcribed spacer region from isolate Pseudozyma sp. JCC207 showed 98% similarity with those of Pseudozyma rugulosa and Pseudozyma aphidis, which are close relatives of the isolate. In considering use of Pseudozyma sp. JCC207 for squalene production, the efficiency of squalene production was investigated under different conditions. Glucose was the best carbon source for the production of squalene. In the presence of yeast extract, squalene production was activated and an optimum ratio of glucose to yeast extract was 4.5. For the optimal squalene production, the concentration of glucose was 40 g l−1 and the best nitrogen source was sodium nitrogen. Pseudozyma sp. JCC207 was shown to produce up to 5.20 g/L of biomass and 340.52 mg/L of squalene. In an optimal condition, the content and yield of squalene produced by Pseudozyma sp. JCC207 were much greater than those obtained from microorganisms previously reported as squalene producers. We identified, classified, and characterized Pseudozyma sp. JCC207 as a novel squalene producer. The squalene production rate of Pseudozyma sp. JCC207 makes it an ideal candidate for the commercialization of microbial squalene.

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References

  • Aguilera Y, Dorado ME, Prada FA, Martínez JJ, Quesada A, Gutiérrez VR (2005) The protective role of squalene in alcohol damage in the chick embryo retina. Exp Eye Res 80:535–543

    Article  CAS  Google Scholar 

  • Avis TJ, Bélanger RR (2002) Mechanisms and means of detection of biocontrol activity of Pseudozyma yeasts against plant-pathogenic fungi. FEMS Yeast Res 2:5–8

    CAS  PubMed  Google Scholar 

  • Begerow D, Bauer R (2000) Phylogenetic placements of ustilaginomycetous anamorphs as deduced from nuclear LSU rDNA sequences. Mycol Res 104(1):53–60

    Article  CAS  Google Scholar 

  • Bhattacharjee P, Shukla VB, Singhal RS, Kulkarni PR (2001) Studies on fermentative production of squalene. World J Microbiol Biotechnol 17:811–816

    Article  CAS  Google Scholar 

  • Boekhout T (1995) Pseudozyma Bandoni emend. Boekhout, a genus for yeast-like anamorphs of Ustilaginales. J Gen Appl Microbiol 41:359–366

    Article  CAS  Google Scholar 

  • Boekhout T, Fell JW (1998) Pseudozyma Bandoni emend. Boekhout and a comparison with the yeast state of Ustilago maydis (De Candolle) Corda. In: Kurtzman CP, Fell JW (eds) The yeasts, a taxonomic study. 4th edn. Elsevier, Amsterdam, pp 790–797

    Chapter  Google Scholar 

  • Chan P, Tomlinson B, Lee CB, Lee YS (1996) Effectiveness and safety of low-dose pravastatin and squalene, alone and in combination, in elderly patients with hypercholesterolemia. J Clin Pharmacol 36:422–427

    Article  CAS  Google Scholar 

  • Dyeberg J, Bang HO (1982) A hypothesis on the development of acute myocardial infarction in Greenlanders. Scand J Clin Lab Invest Suppl 161:7–13

    Article  Google Scholar 

  • Fell JW, Boekhout T, Fonseca A, Scorzetti G, Statzell-Tallman A (2000) Biodiversity and systematics of basidiomycetous yeasts as determined by large subunit rDNA D1/D2 domain sequences analysis. Int J Syst Evol Microbiol 50:1351–1371

    Article  CAS  Google Scholar 

  • Golubev WI, Pfeiffer I, Golubeva EW (2006) Mycocin production in Pseudozyma tsukubaensis. Mycopathologia 162:313–316

    Article  CAS  Google Scholar 

  • Grayburn WS, Collins GB, Hildebrand DF (1992) Fatty acid alteration by a delta 9 desaturase in transgenic tobacco tissue. Biotechnology 10(6):675–678

    CAS  PubMed  Google Scholar 

  • Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57(2–3):267–272

    Article  CAS  Google Scholar 

  • Jiang Y, Fan KW, Wong RT, Chen F (2004) Fatty acid composition and squalene content of the marine microalga Schizochytrium mangrovei. J Agric Food Chem 52(5):1196–1200

    Article  CAS  Google Scholar 

  • Kamimura H, Koga N, Oguri K, Yoshimura H (1992) Enhanced elimination of theophylline, phenobarbital and strychnine from the bodies of rats and mice by squalene treatment. J Phamacobiodyn 15(5):215–221

    Article  CAS  Google Scholar 

  • Kitamoto D, Isoda H, Nakahara T (2002) Functions and potential applications of glycolipid biosurfactants—from energy-saving materials to gene delivery carriers. J Biosci Bioeng 94:187–201

    Article  CAS  Google Scholar 

  • Kohno Y, Egawa Y, Itoh S, Nagaoka S, Takahashi M, Mukai K (1995) Kinetic study of quenching reaction of singlet oxygen and scavenging reaction of free radical by squalene in n-butanol. Biochim Biophys Acta 1256(1):52–56

    Article  Google Scholar 

  • Kulakovskaya TV, Kulakovskaya EV, Golubev WI (2003) ATP leakage from yeast cells treated by extracellular glycolipids of Pseudozyma fusiformata. FEMS Yeast Res 3:401–404

    Article  CAS  Google Scholar 

  • Lewis TE, Nichols PD, McMeekin TA (2001) Sterol and squalene content of a docosahexaenoic acid producing Thraustochytrid: influence of culture age, temperature, and dissolved oxygen. Mar Biotechnol 3:439–447

    Article  CAS  Google Scholar 

  • Mimee B, Labbé C, Pelletier R, Bélanger RR (2005) Antifungal activity of flocculosin, a novel glycolipid isolated from Pseudozyma flocculosa. Antimicrob Agents Chemot 49:1597–1599

    Article  CAS  Google Scholar 

  • Moncalvo JM, Rehner SA, Vilgalys R (1993) Systematics of Lyophyllum section Difformia based on evidence from culture studies and ribosomal DNA sequences. Mycologia 85:788–794

    Article  CAS  Google Scholar 

  • Morita T, Konishi M, Fukuoka T, Imura T Kitamoto D (2006) Discovery of Pseudozyma rugulosa NBRC 10877 as a novel producer of the glycolipid biosurfactants, mannosylerythritol lipids, based on rDNA sequences. Appl Microbiol Biotechnol 73:305–313

    Article  CAS  Google Scholar 

  • Murakoshi M, Nishino H, Tokuda H, Iwashima A, Okuzumi J, Kitano H, Iwasaki R (1992) Inhibition by squalene of the tumor-promoting activity of 12-O-tetradecanoylphorbol-13-acetate in mouse-skin carcinogenesis. Int J Cancer 1992 52(6):950–952

    CAS  Google Scholar 

  • Newmark HL (1999) Squalene, olive oil, and cancer risk: review and hypothesis. Ann NY Acad Sci 889:193–203

    Article  CAS  Google Scholar 

  • Rao CV, Newmark HL, Reedy BS (1998) Chemopreventive effect of squalene on colon cancer. Carcinogenesis 19:287–290

    Article  CAS  Google Scholar 

  • Rau U, Nguyen LA, Schulz S, Wray V, Nimtz M, Roeper H, Koch H, Lang S (2005) Formation and analysis of mannosylerythritol lipids secreted by Pseudozyma aphidis. Appl Microbiol Biotechnol 66(5):551–559

    Article  CAS  Google Scholar 

  • Smith TJ (2000) Squalene: potential chemopreventive agent. Exp Opin Invest Drugs 9(8):1841–1848

    Article  CAS  Google Scholar 

  • Smith TJ, Yang GY, Seril DN, Liao Q, Kim S (1998) Inhibition of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis by dietary olive oil and squalene. Carcinogenesis 19:703–706

    Article  CAS  Google Scholar 

  • Storelli MM, Ceci E, Storelli A, Marcotrigiano GO (2003) Polychlorinated biphenyl, heavy metal and methylmercury residues in hammerhead sharks: contaminant status and assessment. Mar Pollut Bull 46:1035–1039

    Article  CAS  Google Scholar 

  • Storm HM, Oh SY, Kimler BF, Norton S (1993) Radioprotection of mice by dietary squalene. Lipids 28(6):555–559

    Article  CAS  Google Scholar 

  • Sugita T, Takashima M, Poonwan N, Mekha N, Malaithao K, Thungmuthasawat B, Prasarn S, Luangsook P, Kudo T (2003) The first isolation of ustilaginomycetous anamorphic yeasts, Pseudozyma species, from patients’ blood and a description of two new species: P. parantarctica and P. thailandica. Microbiol Immunol 47(3):183–190

    Article  CAS  Google Scholar 

  • Turoczy NJ, Laurenson LJ, Allinson G, Nishikawa M, Lambert DF, Smith C, Cottier JP, Irvine SB, Stagnitti F (2000) Observations on metal concentrations in three species of shark (Deania calcea, Centroscymnus crepidater, and Centroscymnus owstoni) from southeastern Australian waters. J Agric Food Chem 48:4357–4364

    Article  CAS  Google Scholar 

  • Vilgalys R, Hester M (1990) Rapid genetic identification and mapping enzymatically amplified ribosomal DNA from several Cryptococcus species. J Bacteriol 172(8):4238–4246

    Article  CAS  Google Scholar 

  • Wang QM, Jia JH, Bai FY (2006) Pseudozyma hubeiensis sp. nov. and Pseudozyma shanxiensis sp. nov., novel ustilaginomycetous anamorphic yeast species from plant leaves. Int J Syst Evol Microbiol 56:289–293

    Article  CAS  Google Scholar 

  • Ward OP, Singh A (2005) Omega-3/6 fatty acids: alternative sources of production. Process Biochem 40:3627–3652

    Article  CAS  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols, a guide to methods and applications. Academic, New York, pp 315–322

    Google Scholar 

  • Yarrow D (1998) Methods for the isolation, maintenance, classification and identification of yeast. In: Kurtzman CP, Fell JW (eds) The yeasts, a taxonomic study. 4th edn. Elsevier, Amsterdam, pp 75–200

    Google Scholar 

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Acknowledgements

We thank Dr. So Young Kim (Chosun University) and Dr. Yun Cho Chung (Center for University-wide Research Facilities) for critical advice regarding the phylogenetic analysis of the rDNA and the analysis of squalene, respectively. This study was supported by the Research Center for Industrial Development of BioFood Materials at Chonbuk National University, Chonju, Korea.

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Authors and Affiliations

  1. Department of Microbiology, Chonbuk National University Medical School, Chonju, Chonbuk, 561-180, Republic of Korea

    Mi-Hee Chang & Seong-Chool Hong

  2. BDRD Institute, Jinis Biopharmaceuticals Co., 452-32, Jang-Dong, Chonju, Chonbuk, 561-360, Republic of Korea

    Hyeon-Jin Kim

  3. Division of Biological Sciences and Basic Sciences Research Institute, Chonbuk National University, Chonju, Chonbuk, 561-756, Republic of Korea

    Kwang-Yeop Jahng

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  1. Mi-Hee Chang
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  2. Hyeon-Jin Kim
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Correspondence to Seong-Chool Hong.

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Chang, MH., Kim, HJ., Jahng, KY. et al. The isolation and characterization of Pseudozyma sp. JCC 207, a novel producer of squalene. Appl Microbiol Biotechnol 78, 963–972 (2008). https://doi.org/10.1007/s00253-008-1395-4

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  • DOI: https://doi.org/10.1007/s00253-008-1395-4

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