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Efficient production of mannosylerythritol lipids with high hydrophilicity by Pseudozyma hubeiensis KM-59

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Abstract

Mannosylerythritol lipids (MELs) are one of the most promising biosurfactants known because of their multifunctionality and biocompatibility. A previously isolated yeast strain, Pseudozyma sp. KM-59, mainly produced a hydrophilic MEL, namely MEL-C (4-O-[4′-O-acetyl-2′,3′-di-O-alka(e)noyl-β-d-mannopyranosyl]-d-erythritol). In this study, we taxonomically characterize the strain in detail and investigate the culture conditions. The genetic, morphological, and physiological characteristics of the strain coincided well with those of Pseudozyma hubeiensis. On batch culture for 4 days under optimal conditions, the yield of all MELs was 21.8 g/l; MEL-C comprised approximately 65% of the all MELs. Consequently, on fed-batch culture for 16 days, the yield reached 76.3 g/l; the volumetric productivity was approximately 4.8 g l−1 day−1. We further examined the surface-active and self-assembling properties of the hydrophilic MELs produced by the yeast strain. They showed higher emulsifying activities against soybean oil and a mixture of hydrocarbons (2-methylnaphtarene and hexadecane, 1:1) than the synthetic surfactants tested. On water penetration scans, they efficiently formed lyotropic liquid crystalline phases such as myelines and lamella (\( {\text{L}}_{\alpha } \)) in a broad range of their concentrations, indicating higher hydrophilicity than conventional MELs. More interestingly, there was little difference in the liquid crystal formation between the crude product and purified MEL-C. The present glycolipids with high hydrophilicity are thus very likely to have practical potential without further purification and to expand the application of MELs especially their use in washing detergents and oil-in-water-type emulsifiers.

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References

  • Barnett JA, Payne RW, Yarrow D (2000) Yeasts: characteristics and identification, 3rd edn. Cambridge University Press, Cambridge

    Google Scholar 

  • Boekhout T, Nakase T (1998) Pseudozyma Bandoni emend. Boekout and a comparison with the yeast state of Ustilago maydis (de candolle) corda. In: Kurtzman CP, Fell JW (eds) The yeasts a taxonomic study. Elsevier, Amsterdam, Netherlands, pp 790–797

    Chapter  Google Scholar 

  • Imura T, Yanagishita H, Kitamoto D (2004) Coacervate formation from natural glycolipid: one acetyl group on the headgroup triggers coacervate-to-vesicle transition. J Am Chem Soc 126:10804–10805

    Article  CAS  Google Scholar 

  • Imura T, Yanagishita H, Ohira J, Sakai H, Abe M, Kitamoto D (2005) Thermodynamically stable vesicle formation from glycolipid biosurfactant sponge phase. Colloids Surf B Biosurfaces 43:114–121

    Google Scholar 

  • Imura T, Ohta N, Inoue K, Yagi H, Negishi H, Yanagishita H, Kitamoto D (2006) Naturally engineered glycolipid biosurfactants leading to distinctive self-assembled structures. Chem Eur J 12:2434–2440

    Article  CAS  Google Scholar 

  • Imura T, Ito S, Azumi R, Yanagishita H, Sakai H, Abe M, Kitamoto D (2007a) Monolayers assembled from a glycolipid biosurfactant from Pseudozyma (Candida) antarctica serve as a high-affinity ligand system for immunoglobulin G and M. Biotechnol Lett 29:865–870

    Article  CAS  Google Scholar 

  • Imura T, Hikosaka Y, Worakitkanchanakul W, Sakai H, Abe M, Konishi M, Minamikawa H, Kitamoto D (2007b) Aqueous-phase behavior of natural glycolipid biosurfactants mannosylerythritol lipid A: sponge, cubic, and lamella phases. Langmuir 23:1659–1663

    Article  CAS  Google Scholar 

  • Isoda H, Shinmoto H, Kitamoto D, Matsumura M, Nakahara T (1997) Differentiation of human promyelocytic leukemia cell line HL60 by microbial extracellular glycolipids. Lipids 32:263–271

    Article  CAS  Google Scholar 

  • Ito S, Imura T, Fukuoka T, Morita T, Sakai H, Abe M, Kitamoto D (2007) Kinetic studies on the interactions between glycolipid biosurfactants assembled monolayers and various classes of immunoglobulins using surface plasmon resonance. Colloid Surf B 58:165–171

    Article  CAS  Google Scholar 

  • Kitamoto D, Haneishi K, Nakahara T, Tabuchi T (1990) Production of mannosylerythritol lipids by Candida antarctica from vegetable oil. Agric Biol Chem 54:37–40

    CAS  Google Scholar 

  • Kitamoto D, Yanagishita H, Shinbo T, Nakane T, Kamisawa C, Nakahara T (1993) Surface active properties and antimicrobial activities of mannosylerythritol lipids as biosurfactants produced by Candida antarctica. J Biotechnol 29:91–96

    Article  CAS  Google Scholar 

  • Kitamoto D, Ghosh S, Ourisson G, Nakatani Y (2000) Formation of giant vesicles from diacylmannosylerythritols, and their binding to concanavalin A. Chem Commun 10:861–862

    Article  Google Scholar 

  • Kitamoto D, Ikegami T, Suzuki GT, Sasaki A, Takeyama Y, Idemoto Y, Koura N, Yanagishita H (2001) Microbial conversion of n-alkanes into glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma (Candida antarctica). Biotechnol Lett 23:1709–1714

    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 

  • Konishi M, Imura T, Morita T, Fukuoka T, Kitamoto D (2007a) A yeast glycolipid biosurfactant, mannosyl-erythritol lipid, shows high binding affinity towards lectins on a self-assembled monolayer system. Biotechnol Lett 29:473–480

    Article  CAS  Google Scholar 

  • Konishi M, Morita T, Fukuoka T, Imura T, Kakugawa, K, Kitamoto D (2007b) Production of different types of mannosylerythritol lipids as biosurfactants by the newly isolated yeast strains belonging to the genus Pseudozyma. Appl Microbiol Biotechnol 75:521–531

    Article  CAS  Google Scholar 

  • Lang S (2001) Biological amphiphiles (microbial biosurfactants). Curr Opin Colloids Interface Sci 7:11–20

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Morita T, Konishi M, Fukuoka T, Imura T, Kitamoto H, Kitamoto D (2007) Characterization of the genus Pseudozyma by the formation of glycolipid biosurfactants, mannosylerythritol lipids. FEMS Yeast Res 7:286–292

    Article  CAS  Google Scholar 

  • Nakashima T, Shimizu M, Kukizaki M (2000) Particle control of emulsion by membrane emulsification and its application. Adv Drug Deliv Rev 45:47–56

    Article  CAS  Google Scholar 

  • O’Donnell K (1993) Fusarium and its near relatives. In: Reynolds DR, Taylor JW (eds) The fungal holomorph: mitotic, meiotic and pleomorphic speciation in fungal systematics. CAB International, Wallingford, UK, pp 225–233

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Rau U, Nguyen LA, Roeper H, Koch H, Lang S (2005b) Fed-batch bioreactor production of mannosylerythritol lipids secreted by Pseudozyma aphidis. Appl Microbiol Biotechnol 68:607–613

    Article  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTALW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting. Position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  CAS  Google Scholar 

  • Wakamatsu Y, Zhao X, Jin C, Day N, Shibahara M, Nomura N, Nakahara T, Murata T, Yokoyama KK (2001) Mannosylerythritol lipid induces characteristics of neuronal differentiation in PC12 cells through an ERK-related signal cascade. Eur J Biochem 268:374–383

    Article  CAS  Google Scholar 

  • Wang Q-M, Jia J-H, Bai F-Y (2006) Pseudozyma hubeiensis sp. 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 

  • Zeikus JG, Jain MK, Elankovan P (1999) Biotechnology of succinic acid production and markets for derived industrial products. Appl Microbiol Biotechnol 51:545–552

    Article  CAS  Google Scholar 

  • Zhao XX, Murata T, Ohno S, Day N, Song J, Nomura N, Nakahara T, Yokoyama KK (2001) Protein kinase C alpha plays a critical role in mannosylerythritol lipid-induced differentiation of melanoma B16 cells. J Biol Chem 276:39903–39910

    Article  CAS  Google Scholar 

Download references

Acknowledgment

The authors thank Ms. Sugimura, a fellow of the Japan Industrial Technology Association, for her technical assistance. This work was supported by the Industrial Technology Research Grant Program in 06A17501c from the New Energy and Industrial Technology Development Organization (NEDO) of Japan.

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Correspondence to Dai Kitamoto.

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Konishi, M., Morita, T., Fukuoka, T. et al. Efficient production of mannosylerythritol lipids with high hydrophilicity by Pseudozyma hubeiensis KM-59. Appl Microbiol Biotechnol 78, 37–46 (2008). https://doi.org/10.1007/s00253-007-1292-2

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  • DOI: https://doi.org/10.1007/s00253-007-1292-2

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