Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Metabolic Engineering of Saccharomyces cerevisiae to Improve Glucan Biosynthesis

  • Zhou, Xing (The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University) ;
  • He, Jing (The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University) ;
  • Wang, Lingling (The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University) ;
  • Wang, Yang (The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University) ;
  • Du, Guocheng (The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University) ;
  • Kang, Zhen (The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University)
  • Received : 2018.12.26
  • Accepted : 2019.03.25
  • Published : 2019.05.28
Download PDF
⟨ Previous Next ⟩

Abstract

${\beta}$-Glucan is a chief structural polymer in the cell wall of yeast. ${\beta}$-Glucan has attracted intensive attention because of its wide applications in health protection and cosmetic areas. In the present study, the ${\beta}$-glucan biosynthesis pathway in S. Cerevisiae was engineered to enhance ${\beta}$-glucan accumulation. A newly identified bacterial ${\beta}-1$, 6-glucan synthase GsmA from Mycoplasma agalactiae was expressed, and increased ${\beta}$-glucan content by 43%. In addition, other pathway enzymes were investigated to direct more metabolic flux towards the building of ${\beta}$-glucan chains. We found that overexpression of Pgm2 (phosphoglucomutase) and Rho1 (a GTPase for activating glucan synthesis) significantly increased ${\beta}$-glucan accumulation. After further optimization of culture conditions, the ${\beta}$-glucan content was increased by 53.1%. This study provides a new approach to enhance ${\beta}$-glucan biosynthesis in Saccharomyces cerevisiae.

Keywords

References

  1. Barsanti L, Passarelli V, Evangelista V, Frassanito AM, Gualtieri P. 2011. Chemistry, physico-chemistry and applications linked to biological activities of beta-glucans. Nat. Prod. Rep. 28: 457-466. https://doi.org/10.1039/c0np00018c
  2. Samuelsen ABC, Jurgen S, Knutsen SH. 2014. Effects of orally administered yeast-derived beta-glucans: a review. Mol. Nutr. Food Res. 58: 183-193. https://doi.org/10.1002/mnfr.201300338
  3. Kayali H, Ozdag MF, Kahraman S, Aydin A, Gonul E, Sayal A, et al. 2005. The antioxidant effect of ${\beta}$-glucan on oxidative stress status in experimental spinal cord injury in rats. Neurosurg. Rev. 28: 298-302. https://doi.org/10.1007/s10143-005-0389-2
  4. Du B, Bian Z, Xu B. 2014. Skin health promotion effects of natural beta-glucan derived from cereals and microorganisms:a review. Phytother. Res. 28: 159-166. https://doi.org/10.1002/ptr.4963
  5. Dalonso N, Goldman GH, Gern RMM. 2015. ${\beta}-(1{\rightarrow}3),(1{\rightarrow}6)$-Glucans: medicinal activities, characterization, biosynthesis and new horizons. Appl. Microbiol. Biotechnol. 99: 7893-7906. https://doi.org/10.1007/s00253-015-6849-x
  6. Rieder A, Ballance S, Bocker U, Knutsen S. 2017. Quantification of 1,3-${\beta}$-D-glucan from yeast added as a functional ingredient to bread. Carbohydr. Polym. 181: 34. https://doi.org/10.1016/j.carbpol.2017.09.044
  7. Yamaguchi M, Namiki Y, Okada H, Mori Y, Furukawa H, Wang J, et al. 2011. Structome of Saccharomyces cerevisiae determined by freeze-substitution and serial ultrathinsectioning electron microscopy. J. Electron Microsc. 60: 321.
  8. Teparic R, Mrsa V. 2013. Proteins involved in building, maintaining and remodeling of yeast cell walls. Curr. Genet. 59: 171-185. https://doi.org/10.1007/s00294-013-0403-0
  9. Aimanianda V, Clavaud C, Simenel C, Fontaine T, Delepierre M, Latge JP. 2009. Cell wall beta-(1,6)-glucan of Saccharomyces cerevisiae: structural characterization and in situ synthesis. J. Biol. Chem. 284: 13401-13412. https://doi.org/10.1074/jbc.M807667200
  10. Lesage G, Bussey H. 2006. Cell wall assembly in Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 70: 317-343. https://doi.org/10.1128/MMBR.00038-05
  11. Latge JP. 2007. The cell wall: a carbohydrate armour for the fungal cell. Mol. Microbiol. 66: 279-290. https://doi.org/10.1111/j.1365-2958.2007.05872.x
  12. Klis FM, Boorsma A, De Groot PW. 2006. Cell wall construction in Saccharomyces cerevisiae. Yeast 23: 185-202. https://doi.org/10.1002/yea.1349
  13. Levin DE. 2011. Regulation of cell wall biogenesis in Saccharomyces cerevisiae: the cell wall integrity signaling pathway. Genetics 189: 1145-1175. https://doi.org/10.1534/genetics.111.128264
  14. Xu S, Z hang GY, Z hang H, Kitajima T, Nakanishi H, G ao XD. 2016. Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. Microb. Cell Fact. 15: 179. https://doi.org/10.1186/s12934-016-0575-7
  15. Chavan M, Suzuki T, Rekowicz M, Lennarz W. 2003. Genetic, biochemical, and morphological evidence for the involvement of N-glycosylation in biosynthesis of the cell wall beta1,6-glucan of Saccharomyces cerevisiae. Proc.Natl. Acad. Sci. USA 100: 15381-15386. https://doi.org/10.1073/pnas.2536561100
  16. Guan B, Lei J, Su S, Chen F, Duan Z, Chen Y, et al. 2012. Absence of Yps7p, a putative glycosylphosphatidylinositollinked aspartyl protease in Pichia pastoris, results in aberrant cell wall composition and increased osmotic stress resistance. FEMS Yeast Res. 12: 969-979. https://doi.org/10.1111/1567-1364.12002
  17. Wang J, Li M, Zheng F, Niu C, Liu C, Li Q, et al. 2018. Cell wall polysaccharides: before and after autolysis of brewer's yeast. World J. Microbiol. Biotechnol. 34: 137. https://doi.org/10.1007/s11274-018-2508-6
  18. Gaurivaud P, Baranowski E, PauRoblot C, Sagne E, Citti C, Tardy F. 2016. Mycoplasma agalactiae secretion of ${\beta}-(1{\rightarrow}6)$-glucan, a rare polysaccharide in prokaryotes, is governed by high-frequency phase variation. Appl. Environ. Microbiol. 82:AEM.00274-00216.
  19. Dimopoulou M, Vuillemin M, Campbell-Sills H, Lucas PM, Ballestra P, Miot-Sertier C, et al. 2014. Exopolysaccharide (EPS) synthesis by Oenococcus oeni: from genes to phenotypes. PLoS One 9: e98898. https://doi.org/10.1371/journal.pone.0098898
  20. Hrmova M, Stone BA, Fincher GB. 2010. High-yield production, refolding and a molecular modelling of the catalytic module of (1,3)-${\beta}$-d-glucan (curdlan) synthase from Agrobacterium sp. Glycoconj. J. 27: 461-476. https://doi.org/10.1007/s10719-010-9291-4
  21. Liu J J, Zhang GC, Kong, II, Yun E J, Zheng JQ, Kweon DH, et al. 2018. A mutation in PGM2 causing inefficient galactose metabolism in the probiotic yeast Saccharomyces boulardii. Appl. Environ. Microbiol. 84(10): pii: e02858-17.
  22. Bro C, Knudsen S, Regenberg B, Olsson L, Nielsen J. 2005. Improvement of galactose uptake in Saccharomyces cerevisiae through overexpression of phosphoglucomutase: example of transcript analysis as a tool in inverse metabolic engineering. Appl. Environ. Microbiol. 71: 6465-6472. https://doi.org/10.1128/AEM.71.11.6465-6472.2005
  23. Garcia Sanchez R, Hahn-Hagerdal B, Gorwa-Grauslund MF. 2010. PGM2 overexpression improves anaerobic galactose fermentation in Saccharomyces cerevisiae. Microb. Cell Fact. 9: 40. https://doi.org/10.1186/1475-2859-9-40
  24. Lee K S, Hong ME, Jung SC, Ha SJ, Yu BJ, Koo HM, et al. 2011. Improved galactose fermentation of Saccharomyces cerevisiae through inverse metabolic engineering. Biotechnol. Bioeng. 108: 621-631. https://doi.org/10.1002/bit.22988
  25. Eitzen G, Log an MR. 2012. A nalysis of Rho GTPase activation in Saccharomyces cerevisiae. Methods Mol. Biol. 827:369-380. https://doi.org/10.1007/978-1-61779-442-1_24
  26. Auesukaree C. 2017. Molecular mechanisms of the yeast adaptive response and tolerance to stresses encountered during ethanol fermentation. J. Biosci. Bioeng. 124: 133-142. https://doi.org/10.1016/j.jbiosc.2017.03.009
  27. Bleoanca I, Silva AR, Pimentel C, Rodrigues-Pousada C, Menezes Rde A. 2013. Relationship between ethanol and oxidative stress in laboratory and brewing yeast strains. J. Biosci. Bioeng. 116: 697-705. https://doi.org/10.1016/j.jbiosc.2013.05.037
  28. Gibson BR, Lawrence SJ, Leclaire JP, Powell CD, Smart KA. 2007. Yeast responses to stresses associated with industrial brewery handling. FEMS Microbiol. Rev. 31: 535-569. https://doi.org/10.1111/j.1574-6976.2007.00076.x
  29. Sikorski RS, Hieter P. 1989. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19-27. https://doi.org/10.1093/genetics/122.1.19

Cited by

  1. Revealing the Potential of Lipid and β-Glucans Coproduction in Basidiomycetes Yeast vol.8, pp.7, 2019, https://doi.org/10.3390/microorganisms8071034
  2. Yeast Synthetic Biology for the Production of Lycium barbarum Polysaccharides vol.26, pp.6, 2019, https://doi.org/10.3390/molecules26061641
  3. Improved production of β-glucan by a T-DNA-based mutant of Aureobasidium pullulans vol.105, pp.18, 2019, https://doi.org/10.1007/s00253-021-11538-x