Isolation of indigenous Glutathione producing Saccharomyces cerevisiae strains

Document Type: Original Research

Authors

1 Dept. of Microbiology, Faculty of Advanced Science & Technology, Pharmaceutical Science Branch, Islamic Azad University, Tehran, Iran

2 Dept. of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

3 Dept. of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

4 Molecular Microbiology Research Center, Shahed University, Tehran, Iran

Abstract

Background: Glutathione (GSH) is a non-protein thiol compound, which plays an important role in the response to oxidative stress and nutritional stress. The aim of this study was to isolate indigenous S. cerevisiae strains capable of effectively produce GSH.
Methods: One hundred-twenty sweet fruit samples were collected. The strains were isolated on yeast glucose chloramphenicol (YGC) agar medium and identified. The isolates were evaluated for GSH producing on yeast malt (YM) medium. Concentration of glutathione was investigated by recording absorbance of all samples at wavelength 412 nm (Ellman’s method). In addition, optimization of glucose and peptone concentration in culture medium and the effects of various environmental conditions such as temperature (20–35 °C), agitation rate (150–250 rpm), and initial pH (4.0–6.0) were assessed on producing of GSH.
Results: From 120 samples, 80 isolates were identified by morphological, biochemical and molecular tests as S. cerevisiae. Five isolates were capable to produce effectively GSH. The optimal culture conditions were agitation rate, 200 rpm; temperature, 30 °C; initial pH, 6; glucose, 30 g/l; and peptone concentration, 5 g/l. In optimal conditions, the amount of derived glutathione was improved compared to YM basal medium and highest GSH concentration (296.8 mg/l) was obtained after cultivation with shaking for 72 h.
Conclusion: The possibility of obtaining S. cerevisiae cells with a high GSH intracellular content can be considered an interesting opportunity of furthering the range of application and utilization of this molecule.

Keywords

Main Subjects


  1. Lu SC. Glutathione synthesis. BBA-Gen Subjects2013;1830(5):3143-53.
  2. Forman HJ, Zhang H, Rinna A. Glutathione: Overview of its protective roles, measurement, and biosynthesis. Mol Aspects Med 2009;30(1–2):1-12.
  3. Santos LO, Gonzales TA, Úbeda BT, Alegre RM. Influence of culture conditions on glutathione production by Saccharomyces cerevisiae. Appl Microbiol Biotechnol 2007;77(4):763-9.
  4. Zhang T, Wen S, Tan T. Optimization of the medium for glutathione production in Saccharomyces cerevisiae. Process Biochem 2007;42(3):454-8.
  5. Lu SC. Regulation of glutathione synthesis. Mol Aspects Med 2009;30(1):42-59.
  6. Monostori P, Wittmann G, Karg E, Túri S. Determination of glutathione and glutathione disulfide in biological samples: an in-depth review. J Chromatogr B 2009;877(28):3331-46.
  7. Suzuki T, Yokoyama A, Tsuji T, Ikeshima E, Nakashima K, Ikushima S, et al. Identification and characterization of genes involved in glutathione production in yeast. J Biosci Bioeng 2011;112(2):107-13.
  8. Li Y, Wei G, Chen J. Glutathione: a review on biotechnological production. Appl Microbiol Biotechnol 2004;66(3):233-42.
  9. Rollini M, Manzoni M. Influence of different fermentation parameters on glutathione volumetric productivity by Saccharomyces cerevisiae. Process Biochem 2006;41(7):1501-5.
  10. Rollini M, Musatti A, Manzoni M. Production of glutathione in extracellular form by Saccharomyces cerevisiae. Process Biochem 2010;45(4):441-5.
  11. Perricone C, De Carolis C, Perricone R. Glutathione: A key player in autoimmunity. Autoimmun Rev 2009;8(8):697-701.
  12. Xiong Z-Q, Guo M-J, Guo Y-X, Chu J, Zhuang Y-P, Zhang S-L. Efficient extraction of intracellular reduced glutathione from fermentation broth of Saccharomyces cerevisiae by ethanol. Bioresour Technol 2009;100(2):1011-4.
  13. Diosma G, Romanin DE, Rey-Burusco MF, Londero A, Garrote GL. Yeasts from kefir grains: isolation, identification, and probiotic characterization. World J Microbiol Biotechnol 2014;30(1):43-53.
  14. Hampsey M. A review of phenotypes in Saccharomyces cerevisiae. Yeast 1997;13(12):1099-133.
  15. Kurtzman C, Fell JW, Boekhout T. The yeasts: a taxonomic study: Elsevier; 2011.
  16. Josepa S, Guillamon JM, Cano J. PCR differentiation of Saccharomyces cerevisiae from Saccharomyces bayanus/Saccharomyces pastorianus using specific primers. FEMS Microbiol Lett 2000;193(2):255-9.
  17. Altschul SF MT, Scha¨ffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 1997;25:13.
  18. Pastore A, Federici G, Bertini E, Piemonte F. Analysis of glutathione: implication in redox and detoxification. Clin Chim Acta 2003;333(1):19-39.
  19. Wen S, Zhang T, Tan T. Optimization of the amino acid composition in glutathione fermentation. Process Biochem 2005;40(11):3474-9.
  20. Cha J-Y, Park J-C, Jeon B-S, Lee Y-C, Cho Y-S. Optimal fermentation conditions for enhanced glutathione production by Saccharomyces cerevisiae FF-8. J Microbiol 2004;42(1):51-5.
  21. Wen S, Zhang T, Tan T. Utilization of amino acids to enhance glutathione production in Saccharomyces cerevisiae. Enzyme Microb Technol 2004;35(6):501-7.
  22. Wei G, Li Y, Du G, Chen J. Effect of surfactants on extracellular accumulation of glutathione by Saccharomyces cerevisiae. Process Biochem 2003;38(8):1133-8.
  23. Liu C-H, Hwang C-F, Liao C-C. Medium optimization for glutathione production by Saccharomyces cerevisiae. Process Biochem 1999;34(1):17-23.
  24. Liu H, Lin J, Cen P, Pan Y. Co-production of S-adenosyl-L-methionine and glutathione from spent brewer’s yeast cells. Process Biochem 2004;39(12):1993-7.
  25. Penninckx M. A short review on the role of glutathione in the response of yeasts to nutritional, environmental, and oxidative stresses. Enzyme Microb Technol 2000;26(9):737-42.
  26. Suzuki M KM. Effects of ATP level on glutathione regeneration in rabbit and guinea pig erythrocytes. Comp Biochem Physiol B 1992;103:4.
  27. Ubiyvovk VM, Ananin VM, Malyshev AY, Kang HA, Sibirny AA. Optimization of glutathione production in batch and fed-batch cultures by the wild-type and recombinant strains of the methylotrophic yeast Hansenula polymorpha DL-1. BMC Biotechnol 2011;11(1):8.
  28. Udeh K, Achremowicz B. High-glutathione containing yeast Saccharomyces cerevisiae: optimization of production. Acta Microbiol Pol 1996;46(1):105-14.