1887

Abstract

The limited catalytic efficiency of cellulose-degrading enzymes restricts cellulose digestion. We investigated the transcriptional regulation of genes encoding key cellulose degrading enzymes, namely β-glucosidases, in the industrial actinobacterium We observed that the expression of most β-glucosidase-encoding genes was controlled by the availability of nitrogen and phosphate via their respective global regulators, namely GlnR and PhoP. Electrophoretic mobility shift assay demonstrated that GlnR and PhoP bound directly to the promoters of β-glucosidase-encoding genes. Deletion of resulted in lower transcript levels and activity of β-glucosidases, leading to decreased bacterial growth on cellulose. Overexpression of and or nitrogen/phosphate starvation increased the transcript levels and total activity of β-glucosidases. Moreover, GlnR/PhoP-mediated cellobiose utilization was also observed in A3(2). These findings provide insights into the regulatory roles played by GlnR and PhoP in coordinating nitrogen/phosphate metabolism and carbohydrate utilization, and indicate potential strategies for cellulose fermentation in the production of bio-based chemicals by actinobacteria.

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2018-05-01
2024-04-20
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References

  1. Werpy T, Petersen G, Aden A, Bozell J, Holladay J et al. Top value added chemicals from biomass. In Results of Screening for Potential Candidates from Sugars and Synthesis Gas vol. 1 Oak Ridge, TN, USA: NREL (2004) U.S. Department of Energy; 2012
    [Google Scholar]
  2. Cherubini F. The biorefinery concept: Using biomass instead of oil for producing energy and chemicals. Energy Convers Manag 2010; 51:1412–1421 [View Article]
    [Google Scholar]
  3. Sørensen A, Lübeck M, Lübeck PS, Ahring BK. Fungal beta-glucosidases: a bottleneck in industrial use of lignocellulosic materials. Biomolecules 2013; 3:612–631 [View Article][PubMed]
    [Google Scholar]
  4. Lynd LR, Weimer PJ, van Zyl WH, Pretorius IS. Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 2002; 66:506–577 [View Article][PubMed]
    [Google Scholar]
  5. Binder JB, Raines RT. Fermentable sugars by chemical hydrolysis of biomass. Proc Natl Acad Sci USA 2010; 107:4516–4521 [View Article][PubMed]
    [Google Scholar]
  6. Mosier N, Wyman C, Dale B, Elander R, Lee YY et al. Features of promising technologies for pretreatment of lignocellulosic biomass. Bioresour Technol 2005; 96:673–686 [View Article][PubMed]
    [Google Scholar]
  7. Xu Q, Singh A, Himmel ME. Perspectives and new directions for the production of bioethanol using consolidated bioprocessing of lignocellulose. Curr Opin Biotechnol 2009; 20:364–371 [View Article][PubMed]
    [Google Scholar]
  8. Lee WH, Nan H, Kim HJ, Jin YS. Simultaneous saccharification and fermentation by engineered Saccharomyces cerevisiae without supplementing extracellular β-glucosidase. J Biotechnol 2013; 167:316–322 [View Article][PubMed]
    [Google Scholar]
  9. Chauve M, Mathis H, Huc D, Casanave D, Monot F et al. Comparative kinetic analysis of two fungal β-glucosidases. Biotechnol Biofuels 2010; 3:3 [View Article][PubMed]
    [Google Scholar]
  10. Martins LF, Kolling D, Camassola M, Dillon AJ, Ramos LP. Comparison of Penicillium echinulatum and Trichoderma reesei cellulases in relation to their activity against various cellulosic substrates. Bioresour Technol 2008; 99:1417–1424 [View Article][PubMed]
    [Google Scholar]
  11. Olofsson K, Bertilsson M, Lidén G. A short review on SSF - an interesting process option for ethanol production from lignocellulosic feedstocks. Biotechnol Biofuels 2008; 1:7 [View Article][PubMed]
    [Google Scholar]
  12. Liu ZL, Weber SA, Cotta MA, Li SZ. A new β-glucosidase producing yeast for lower-cost cellulosic ethanol production from xylose-extracted corncob residues by simultaneous saccharification and fermentation. Bioresour Technol 2012; 104:410–416 [View Article][PubMed]
    [Google Scholar]
  13. Spindler DD, Wyman CE, Grohmann K, Mohagheghi A. Simultaneous saccharification and fermentation of pretreated wheat straw to ethanol with selected yeast strains and β-glucosidase supplementation. Appl Biochem Biotechnol 1989; 20-21:529–540 [View Article]
    [Google Scholar]
  14. Lynd LR, van Zyl WH, McBride JE, Laser M. Consolidated bioprocessing of cellulosic biomass: an update. Curr Opin Biotechnol 2005; 16:577–583 [View Article][PubMed]
    [Google Scholar]
  15. Guo ZP, Zhang L, Ding ZY, Gu ZH, Shi GY. Development of an industrial ethanol-producing yeast strain for efficient utilization of cellobiose. Enzyme Microb Technol 2011; 49:105–112 [View Article][PubMed]
    [Google Scholar]
  16. Shen Y, Zhang Y, Ma T, Bao X, Du F et al. Simultaneous saccharification and fermentation of acid-pretreated corncobs with a recombinant Saccharomyces cerevisiae expressing beta-glucosidase. Bioresour Technol 2008; 99:5099–5103 [View Article][PubMed]
    [Google Scholar]
  17. Van Zyl WH, Lynd LR, den Haan R, McBride JE. Consolidated bioprocessing for bioethanol production using Saccharomyces cerevisiae . Adv Biochem Eng Biotechnol 2007; 108:205–235 [View Article][PubMed]
    [Google Scholar]
  18. Yamada R, Taniguchi N, Tanaka T, Ogino C, Fukuda H et al. Direct ethanol production from cellulosic materials using a diploid strain of Saccharomyces cerevisiae with optimized cellulase expression. Biotechnol Biofuels 2011; 4:8 [View Article][PubMed]
    [Google Scholar]
  19. Guo Z, Duquesne S, Bozonnet S, Cioci G, Nicaud JM et al. Development of cellobiose-degrading ability in Yarrowia lipolytica strain by overexpression of endogenous genes. Biotechnol Biofuels 2015; 8:1–16 [View Article][PubMed]
    [Google Scholar]
  20. Foreman PK, Brown D, Dankmeyer L, Dean R, Diener S et al. Transcriptional regulation of biomass-degrading enzymes in the filamentous fungus Trichoderma reesei . J Biol Chem 2003; 278:31988–31997 [View Article][PubMed]
    [Google Scholar]
  21. Martinez D, Berka RM, Henrissat B, Saloheimo M, Arvas M et al. Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina). Nat Biotechnol 2008; 26:553–560 [View Article][PubMed]
    [Google Scholar]
  22. Nitta M, Furukawa T, Shida Y, Mori K, Kuhara S et al. A new Zn(II)2Cys6-type transcription factor BglR regulates β-glucosidase expression in Trichoderma reesei . Fungal Genet Biol 2012; 49:388–397 [View Article][PubMed]
    [Google Scholar]
  23. Rodríguez H, Fraga R. Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnol Adv 1999; 17:319–339 [View Article][PubMed]
    [Google Scholar]
  24. Martín JF, Sola-Landa A, Santos-Beneit F, Fernández-Martínez LT, Prieto C et al. Cross-talk of global nutritional regulators in the control of primary and secondary metabolism in Streptomyces. Microb Biotechnol 2011; 4:165–174 [View Article][PubMed]
    [Google Scholar]
  25. Yao LL, Ye BC. Reciprocal regulation of GlnR and PhoP in Response to nitrogen and phosphate limitations in Saccharopolyspora erythraea . Appl Environ Microbiol 2016; 82:409–420 [View Article][PubMed]
    [Google Scholar]
  26. Tiffert Y, Supra P, Wurm R, Wohlleben W, Wagner R et al. The Streptomyces coelicolor GlnR regulon: identification of new GlnR targets and evidence for a central role of GlnR in nitrogen metabolism in actinomycetes. Mol Microbiol 2008; 67:861–880 [View Article][PubMed]
    [Google Scholar]
  27. Yao LL, Liao CH, Huang G, Zhou Y, Rigali S et al. GlnR-mediated regulation of nitrogen metabolism in the actinomycete Saccharopolyspora erythraea . Appl Microbiol Biotechnol 2014; 98:7935–7948 [View Article][PubMed]
    [Google Scholar]
  28. Puri-Taneja A, Paul S, Chen Y, Hulett FM. CcpA causes repression of the phoPR promoter through a novel transcription start site, PA6 . J Bacteriol 2006; 188:1266–1278 [View Article][PubMed]
    [Google Scholar]
  29. Rodríguez-García A, Sola-Landa A, Apel K, Santos-Beneit F, Martín JF. Phosphate control over nitrogen metabolism in Streptomyces coelicolor: direct and indirect negative control of glnR, glnA, glnII and amtB expression by the response regulator PhoP. Nucleic Acids Res 2009; 37:3230–3242 [View Article][PubMed]
    [Google Scholar]
  30. Allenby NE, Laing E, Bucca G, Kierzek AM, Smith CP. Diverse control of metabolism and other cellular processes in Streptomyces coelicolor by the PhoP transcription factor: genome-wide identification of in vivo targets. Nucleic Acids Res 2012; 40:9543–9556 [View Article][PubMed]
    [Google Scholar]
  31. Liao CH, Yao L, Xu Y, Liu WB, Zhou Y et al. Nitrogen regulator GlnR controls uptake and utilization of non-phosphotransferase-system carbon sources in actinomycetes. Proc Natl Acad Sci USA 2015; 112:15630–15635 [View Article][PubMed]
    [Google Scholar]
  32. Xu Y, Liao CH, Yao LL, Ye X, Ye BC. GlnR and PhoP directly regulate the transcription of genes encoding starch-degrading, amylolytic enzymes in Saccharopolyspora erythraea . Appl Environ Microbiol 2016; 82:6819–6830 [View Article][PubMed]
    [Google Scholar]
  33. Fink D, Weissschuh N, Reuther J, Wohlleben W, Engels A. Two transcriptional regulators GlnR and GlnRII are involved in regulation of nitrogen metabolism in Streptomyces coelicolor A3(2). Mol Microbiol 2002; 46:331–347 [View Article][PubMed]
    [Google Scholar]
  34. Shewale JG. Beta-Glucosidase: its role in cellulase synthesis and hydrolysis of cellulose. Int J Biochem 1982; 14:435–443 [View Article][PubMed]
    [Google Scholar]
  35. Xiao Z, Zhang X, Gregg DJ, Saddler JN. Effects of sugar inhibition on cellulases and beta-glucosidase during enzymatic hydrolysis of softwood substrates. Appl Biochem Biotechnol 2004; 115:1115–1126[PubMed] [Crossref]
    [Google Scholar]
  36. Murphy L, Bohlin C, Baumann MJ, Olsen SN, Sørensen TH et al. Product inhibition of five Hypocrea jecorina cellulases. Enzyme Microb Technol 2013; 52:163–169 [View Article][PubMed]
    [Google Scholar]
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