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Improvement of Erythrose Reductase Activity, Deletion of By-products and Statistical Media Optimization for Enhanced Erythritol Production from Yarrowia lipolytica Mutant 49

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Abstract

The purpose of the present investigation was to produce erythritol by Yarrowia lipolytica mutant without any by-products. Mutants of Y. lipolytica were generated by ultra-violet for enhancing erythrose reductase (ER) activity and erythritol production. The mutants showing the highest ER activity were screened by triphenyl tetrazolium chloride agar plate assay. Productivity of samples was analyzed by thin-layer chromatography and high-performance liquid chromatography equipped with the refractive index detector. One of the mutants named as mutant 49 gave maximum erythritol production without any other by-products (particularly glycerol). Erythritol production and specific ER activity in mutant 49 increased to 1.65 and 1.47 times, respectively, in comparison with wild-type strain. The ER gene of wild and mutant strains was sequenced and analyzed. A general comparison of wild and mutant gene sequences showed the replacement of Asp270 with Glu270 in ER protein. In order to enhance erythritol production, we used a three component-three level-one response Box–Behnken of response surface methodology model. The optimum medium composition for erythritol production was found to be (g/l) glucose 279.49, ammonium sulfate 9.28, and pH 5.41 with 39.76 erythritol production.

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References

  1. Abe S, Morioka S (1999) Method of producing erythritol. United State patent 5902739

  2. Biro JC (2008) Correlation between nucleotide composition and folding energy of coding sequences with special attention to wobble bases. Theor Biol Med Model 5:14–22

    Article  PubMed Central  PubMed  Google Scholar 

  3. Bradford MM (1976) A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  4. Care S, Bignon C, Pelissier MC, Blanc E, Canard B, Coutard B (2008) The translation of recombinant proteins in E. coli can be improved by in silico generating and screening random libraries of a 270/+96 mRNA region with respect to the translation initiation codon. Nucleic Acids Res 36:1–6

    Article  Google Scholar 

  5. Ghezelbash GR, Nahvi I, Rabani M (2012) Study of polyols production by Yarrowia lipolytica in batch culture and optimization of growth condition for maximum production. Jundishapur J Microbiol 5:546–549

    Article  Google Scholar 

  6. Hajny GJ, Smith JH, Garver JC (1964) Erythritol production by a yeast-like fungus. Appl Microbiol 12:240–246

    CAS  PubMed Central  PubMed  Google Scholar 

  7. Heussen C, Dowdle EB (1980) Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulphate and copolymerized substrates. Anal Biochem 102:196–202

    Article  CAS  PubMed  Google Scholar 

  8. Hirata Y, Igarashi K, Ezaki S, Atomi H, Imanaka T (1999) High-level production of erythritol by strain 618A-01 isolated from pollen. J Biosci Bioeng 87:630–635

    Article  CAS  PubMed  Google Scholar 

  9. Hoffman CS (1997) Current protocols in molecular biology. Wiley, New York

    Google Scholar 

  10. Kim SY, Lee KH, Kim JH, Oh DK (1997) Erythritol production by controlling osmotic pressure in Trigonopsis variabilis. Biotechnol Lett 19:727–729

    Article  CAS  Google Scholar 

  11. Kozak M (2005) Regulation of translation via mRNA structure in prokaryotes and eukaryotes. Gene 361:13–37

    Article  CAS  PubMed  Google Scholar 

  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  13. Laxman SS, Ramchandra VG, Bhalchandra KV, Karthik N (2011) Strain improvement and statistical media optimization for enhanced erythritol production with minimal by-products from Candida magnoliae mutant R23. Biochem Eng J 55:92–100

    Article  Google Scholar 

  14. Lee DH, Lee YJ, Ryu YW, Seo JH (2010) Molecular cloning and biochemical characterization of a novel erythrose reductase from Candida magnoliae JH110. Microb Cell Fact 9:43–55

    Article  PubMed Central  PubMed  Google Scholar 

  15. Lee JK, Ha SJ, Kim SY, Oh DK (2001) Increased erythritol production in Torula sp. with inositol and phytic acid. Biotechnol Lett 23:497–500

    Article  CAS  Google Scholar 

  16. Lee JK, Hong KW, Kim SY (2003) Purification and properties of a NADPH-dependent erythrose reductase from the newly isolated Torula coralline. Biotechnol Prog 19:495–500

    Article  CAS  PubMed  Google Scholar 

  17. Lee JK, Kim SY, Ryu YW, Seo JH, Kim JH (2003) Purification and characterization of a novel erythrose reductase from Candida magnoliae. Appl Environ Microbiol 69:3710–3718

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Lee KJ, Lim JY (2003) Optimized condition for high erythritol production by Penicillium sp. KJ-UV29, mutant of Penicillium sp. KJg81. Biotechnol Bioprocess Eng 8:173–178

    Article  CAS  Google Scholar 

  19. Lin SJ, Wen CY, Liau JC, Chu WS (2001) Screening and production of erythritol by newly isolated osmophilic yeast-like fungi. Process Biochem 36:1249–1258

    Article  CAS  Google Scholar 

  20. Lin SJ, Wen CY, Wang PM, Huang JC, Wei CL, Chang JW, Chu WS (2010) High-level production of erythritol by mutants of osmophilic Moniliella sp. Process Biochem 45:973–979

    Article  CAS  Google Scholar 

  21. Park EH, Lee HY, Ryu YW, Seo JH, Kim MD (2011) Role of osmotic and salt stress in the expression of erythrose reductase in Candida magnoliae. J Microbiol Biotechnol 10:1064–1068

    Article  Google Scholar 

  22. Park JB, Seo BC, Kim JR, Park YK (1998) Production of erythritol in fed-batch cultures of Trichosporon sp. J Ferment Bioeng 86:577–580

    Article  CAS  Google Scholar 

  23. Park YC, Lee DY, Lee DH, Kim HJ, Ryu YW, Seo JH (2005) Proteomics and physiology of erythritol-producing strains. J Chromatogr B 815:251–260

    Article  CAS  Google Scholar 

  24. Patil SV, Jayaraman VK, Kulkarni BD (2002) Optimization of media by evolutionary algorithms for production of polyols. Appl Biochem Biotechnol 102:119–128

    Article  PubMed  Google Scholar 

  25. Rymowicz W, Rywinska A, Marcinkiewicz M (2009) High-yield production of erythritol from raw glycerol in fed-batch cultures of Yarrowia lipolytica. Biotechnol Lett 31:377–380

    Article  CAS  PubMed  Google Scholar 

  26. Shukla P, Garai D, Zafar M, Gupta K, Shrivastava S (2007) Process parameters optimization for lipase production by Rhizopus oryzae kg-10 under submerged fermentation using response surface methodology. J Appl Sci Environ Sanit 2:93–103

    Google Scholar 

  27. Tokuoka K, Ishitani T, Chungz WC (1992) Accumulation of polyols and sugars in some sugar tolerant yeasts. J Gen Appl Microbiol 38:35–46

    Article  CAS  Google Scholar 

  28. Tomaszewska L, Rywinska A, Gładkowski W (2012) Production of erythritol and mannitol by Yarrowia lipolytica yeast in media containing glycerol. J Ind Microbiol Biotechnol 39:1333–1343

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Acknowledgments

The current study was supported by the grant of Postgraduate Administration Office of the University of Isfahan to Gh. R. Ghezelbash for obtaining Ph.D. degree. In addition, the authors are grateful to Sara Ghashghaei and Vahid Niknezhad from University of Isfahan for the helps with experiments.

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Authors and Affiliations

  1. Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran

    Gholam Reza Ghezelbash

  2. Department of Biology, Faculty of Science, University of Isfahan, Isfahan, Iran

    Iraj Nahvi & Rahman Emamzadeh

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  1. Gholam Reza Ghezelbash
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  2. Iraj Nahvi
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  3. Rahman Emamzadeh
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Correspondence to Iraj Nahvi.

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Ghezelbash, G.R., Nahvi, I. & Emamzadeh, R. Improvement of Erythrose Reductase Activity, Deletion of By-products and Statistical Media Optimization for Enhanced Erythritol Production from Yarrowia lipolytica Mutant 49. Curr Microbiol 69, 149–157 (2014). https://doi.org/10.1007/s00284-014-0562-3

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  • DOI: https://doi.org/10.1007/s00284-014-0562-3

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