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Biomass Conversion and Biorefinery

, Volume 8, Issue 1, pp 97–111 | Cite as

Binary- and triple-enzyme cocktails and their application mode affect fermentable sugar release from pretreated lignocellulo-starch biomass

  • M.G. Mithra
  • J. Sreekumar
  • G. PadmajaEmail author
Original Article

Abstract

Lignocellulo-starch biomass (LCSB) comprising roots and vegetable processing wastes has high starch besides cellulose and hemicelluloses and warrants different pretreatment and saccharification approaches. The fermentable sugar yield from steam/dilute sulphuric acid (DSA)-pretreated biomass during saccharification with binary [cellulase + amylolytic enzyme (Stargen)] or triple (cellulase + xylanase + Stargen) enzyme cocktails was compared. The factors such as pH (5.0), temperature (50 °C) and enzyme dosage (16 FPU/g cellulose) for cellulase (Ecozyme RT80) action were optimized using response surface methodology. As pretreated liquor is rich in sugars, whole slurry saccharification was needed for LCSBs and saccharification efficiency (120 h) was significantly higher for steam-pretreated biomass with all application modes. Preferential hydrolysis of starch in steam-pretreated biomass by Stargen followed by cellulolysis was advantageous than the application sequence with cellulase followed by Stargen. Triple-enzyme-based saccharification of steam-pretreated biomass significantly enhanced the overall conversion efficiency (OCE; 85–98%) compared to only 28–49% in the native untreated biomass, while lower OCE was observed in the case of DSA-pretreated and saccharified biomass. Supplementation with both xylanase and Stargen pronouncedly enhanced the OCE for steam-pretreated biomass with only insignificant difference between the exposure periods, indicating the obligatory need for both enzymes for optimal saccharification of LCSBs.

Keywords

Lignocellulo-starch biomass Pretreatment Saccharification Enzyme cocktails Application mode Fermentable sugars 

Notes

Acknowledgements

The authors acknowledge with gratitude the financial support from the Kerala State Council for Science, Technology & Environment (Grant no. 853/2015/KSCSTE) and the facilities provided by the director, ICAR-CTCRI, for the study. Stargen™ 002 was received by courtesy from M/s Danisco US Inc., USA.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13399_2017_237_MOESM1_ESM.docx (224 kb)
ESM 1 (DOCX 223 kb)
13399_2017_237_MOESM2_ESM.docx (224 kb)
ESM 2 (DOCX 223 kb)

References

  1. 1.
    Sarkar N, Ghosh SK, Bannerjee S, Aikat K (2012) Bioethanol production from agricultural wastes: an overview. Renew Ener 37:19–28CrossRefGoogle Scholar
  2. 2.
    Fuglestvedt J, Berntsen T, Myhre G, Rypdal K, Skeie RB (2008) Climate forcing from the transport sectors. Proc Natl Acad Sci U S A 105:454–458CrossRefGoogle Scholar
  3. 3.
    Oghgren KH, Hahn B, Zacchi G (2006) Simultaneous saccharification and co- fermentation of glucose and xylose in steam pretreated corn stover at high fiber content with S. cerevisiae. J Biotech 126:488–496CrossRefGoogle Scholar
  4. 4.
    Wyman CE (1999) Biomass ethanol: technical progress, opportunities and commercial challenges. Annu Rev Energ Environ 24:189–226CrossRefGoogle Scholar
  5. 5.
    Himmel ME, Ding SY, Johnson DK, Adney WS, Nimlos MR, Brady JW, Foust TD (2007) Biomass recalcitrance: engineering plants and enzymes for biofuels production. Science 325:804–807CrossRefGoogle Scholar
  6. 6.
    Yang B, Wyman CE (2008) Pretreatment: the key to unlocking low cost cellulosic ethanol. Biofuels Bioprod Bioref 2:26–40CrossRefGoogle Scholar
  7. 7.
    Alvira P, Tomas-Pejo E, Ballesteros M, Negro MJ (2010) Pretreatment technologies for an efficient bioethanol production process based on enzymatic hydrolysis: a review. Bioresour Technol 101:4851–4861CrossRefGoogle Scholar
  8. 8.
    Hendriks ATWM, Zeeman G (2009) Pretreatments to enhance the digestibility of lignocellulosic biomass. Bioresour Technol 100:10–18CrossRefGoogle Scholar
  9. 9.
    Taherzadeh MJ, Karimi K (2007) Enzyme-based hydrolysis process for ethanol production from lignocellulosic materials: a review. Bioresources 2(4):707–738Google Scholar
  10. 10.
    Leu SY, Zhu JY (2013) Substrate-related factors affecting enzymatic saccharification of lignocelluloses: our recent understanding. Bioener Res 6. doi: 10.1007/s12155-012 9286-1
  11. 11.
    Zhao X, Zhang L, Liu D (2012) Biomass recalcitrance. Part I: the chemical compositions and physical structures affecting the enzymatic hydrolysis of lignocellulose. Biofuels Bioprod Bioref 6(4):465–482CrossRefGoogle Scholar
  12. 12.
    Yang B, Wyman CE (2004) Effect of xylan and lignin removal by batch and flow through pretreatment on the enzymatic digestibility of corn stover cellulose. Biotechnol Bioeng 86(1):88–95CrossRefGoogle Scholar
  13. 13.
    Hu J, Arantes V, Saddler JN (2011) The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: is it an additive or synergistic effect? Biotechnol Biofuels 4:1–13CrossRefGoogle Scholar
  14. 14.
    Zhang C, Zhuang XX, Wang ZJ, Matt F, St. John F, Zhu JY (2013) Xylanase supplementation on enzymatic saccharification of dilute acid pretreated poplars at different severities. Cellulose 20:1937–1946CrossRefGoogle Scholar
  15. 15.
    Moxley G, Gaspar AR, Higgins D, Xu H (2012) Structural changes of corn stover lignin during acid pretreatment. J Indus Microbiol Biotechnol 39(9):1289–1299CrossRefGoogle Scholar
  16. 16.
    Selig MJ, Knoshaug EP, Adney WS, Himmel ME, Decker SR (2008) Synergistic enhancement of cellobiohydrolase performance on pretreated corn stover by addition of xylanase and esterase activities. Bioresour Technol 99:4997–5005CrossRefGoogle Scholar
  17. 17.
    Murashima K, Kosugi A, Doi RH (2003) Synergistic effects of cellulosomal xylanase and cellulases from Clostridium cellulovorans on plant cell wall degradation. J Bacteriol 185(5):1518–1524CrossRefGoogle Scholar
  18. 18.
    Keshwani DR, Cheng JJ (2009) Switchgrass for bioethanol and other value-added applications: a review. Bioresour Technol 100:137–145CrossRefGoogle Scholar
  19. 19.
    Holtzapple MT, Ripley EP, Nikolaou M (1994) Saccharification, fermentation, and protein recovery from low-temperature AFEX-treated coastal Bermuda grass. Biotechnol Bioeng 44:1122–1131CrossRefGoogle Scholar
  20. 20.
    Kim Y, Mosier NS, Ladisch MR (2009) Enzymatic digestion of liquid hot water pretreated hybrid poplar. Biotechnol Prog 25:340–348CrossRefGoogle Scholar
  21. 21.
    Zhu S, Wu Y, Yu Z, Wang C, Yu F et al (2006) Comparison of three microwave/chemical pretreatment processes for enzymatic hydrolysis of rice straw. Biosyst Eng 93:279–283CrossRefGoogle Scholar
  22. 22.
    Xu J, Chen H, Kádár Z, Thomsen AB, Schmidt JE, Peng H (2011) Optimization of microwave pretreatment on wheat straw for ethanol production. Biomass Bioener 35:3859–3864CrossRefGoogle Scholar
  23. 23.
    Benjamin Y, Cheng H, Görgens JF (2014) Optimization of dilute sulphuric acid pretreatment to maximize combined sugar yield form sugarcane bagasse for ethanol production. Appl Biochem Biotechnol 172:610–630CrossRefGoogle Scholar
  24. 24.
    Mithra MG, Padmaja G (2016 a) Compositional profile and ultrastructure of steam and dilute sulfuric acid pretreated root and vegetable processing residues. Curr Biotechnol 7. doi: 10.2174/2211550105666160916124120
  25. 25.
    Mithra MG, Padmaja G (2016 b) Lime pretreatment associated compositional and ultrastructural changes in selected root and vegetable processing residues. Amer J Biomass Bioener (In press).Google Scholar
  26. 26.
    Mithra MG, Padmaja G (2016 c) Comparative alterations in the compositional profile of selected root and vegetable peels subjected to three pretreatments for enhanced saccharification. Ind J Biotechnol (In press).Google Scholar
  27. 27.
    Lin L, Yan R, Liu Y, Jiang W (2010) In-depth investigation of enzymatic hydrolysis of biomass wastes based on three major components: cellulose, hemicelluloses and lignin. Bioresour Technol 101:8217–8223CrossRefGoogle Scholar
  28. 28.
    Saha BC, Cotta MA (2009) Comparison of pretreatment strategies for enzymatic saccharification and fermentation of barley straw to ethanol. New Biotechnol 27:10–16CrossRefGoogle Scholar
  29. 29.
    Divya Nair MP, Padmaja G, Moorthy SN (2011) Biodegradation of cassava starch factory residue using a combination of cellulases, xylanases and hemicellulases. Biomass Bioener 35:1211–1218CrossRefGoogle Scholar
  30. 30.
    Saha BC, Bothast RJ (1999) Pretreatment and enzymatic saccharification of corn fiber. Appl Biochem Biotechnol 76:65–77CrossRefGoogle Scholar
  31. 31.
    Ghose TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268Google Scholar
  32. 32.
    Tomaz T, Roche A (2002) Hydrophobic interaction, chromatography of Trichoderma reesei cellulase on polypropylene glycol-sepharose. Separation Sci Technol 37:1–11CrossRefGoogle Scholar
  33. 33.
    Nelson N (1944) A photometric adaptation of the Somogyi method for determination of glucose. J Biol Chem 153:375–380Google Scholar
  34. 34.
    Anon (2009) STARGEN™ 002: Granular starch hydrolyzing enzyme for ethanol production. Product information published by Genencor International, a Division of Danisco, Danisco US Inc., Available from: http://www.genencor.com. Accessed December 22, 2014.
  35. 35.
    AOAC (1995) Official Methods of Analysis, Sixteenth ed., Washington, DC: Association of Official Analytical Chemists.Google Scholar
  36. 36.
    Box G, Behnken D (1960) Some new three level designs for the study of quantitative variables. Techometrics 2:455–475MathSciNetCrossRefzbMATHGoogle Scholar
  37. 37.
    SAS (2010) Cary NC. USA: SAS Institute Inc.Google Scholar
  38. 38.
    Dien SS, Ximenes EA, O’Brien PJ, Moniruzzaman M, Li X-L, Balan V, Dale B, Cotta MA (2008) Enzyme characterization for hydrolysis of AFEX and liquid hot-water pretreated distillers’ grains and their conversion to ethanol. Bioresour Technol 99:5216–5225CrossRefGoogle Scholar
  39. 39.
    Saha BC, Cotta MA (2010) Comparison of pretreatment strategies for enzymatic saccharification and fermentation of barley straw to ethanol. New Biotechnol 28:10–16CrossRefGoogle Scholar
  40. 40.
    Bothast RJ, Saha BC (1997) Ethanol production from agricultural biomass substrates. Adv Appl Microbiol 44:261–286CrossRefGoogle Scholar
  41. 41.
    Qing Q, Yang B, Wyman CE (2010) Xylo-oligomers are strong inhibitors of cellulose hydrolysis by enzymes. Bioresour Technol 101:9624–9630CrossRefGoogle Scholar
  42. 42.
    Mosier M, Wyman CE, Dale B, Elander R, Lee YY, Holtzapple M, Ladisch M (2005) Features of promising technologies for pretreatment of lignocellulosic biomass. Bioresour Technol 96:673–686CrossRefGoogle Scholar
  43. 43.
    Lamptey J, Robinson CW, Moo-Young M (1985) Enhanced enzymatic hydrolysis of lignocellulosic biomass pretreated by low-pressure steam autohydrolysis. Biotechnol Letters 7:531–536CrossRefGoogle Scholar
  44. 44.
    Mok WSL, Antal MJ (1992) Uncatalyzed solvolysis of whole biomass hemicelluloses by hot compressed liquid water. Ind Eng Chem Res 31:1157–1161CrossRefGoogle Scholar
  45. 45.
    Bisaria VS, Ghose TK (1981) Biodegradation of cellulosic materials: substrate, microorganisms, enzymes and products. Enz Microb Technol 3:90–104CrossRefGoogle Scholar
  46. 46.
    Tejirian A, Fu F (2011) Inhibition of enzymatic cellulolysis by phenolic compounds. Enz Microb Technol 48:239–247CrossRefGoogle Scholar
  47. 47.
    Ximenes EA, Kim Y, Mosier NS, Dien BS, Ladisch MR (2010) Inhibition of cellulases by phenols. Enz Microb Technol 46:170–176CrossRefGoogle Scholar
  48. 48.
    Mithra MG, Padmaja G (2016 d) Phenolic inhibitors of saccharification and fermentation in lignocellulo-starch prehydrolysates and comparative efficacy of detoxification treatments. J Biomass Biofuel 3:1–15. doi: 10.11159/jbb.2016.001 Google Scholar
  49. 49.
    Ko JK, Um Y, Park YC, Seo JH, Kim KH (2015) Compounds inhibiting the bioconversion of hydrothermally pretreated lignocellulose. Appl Microbiol Biotechnol 99:4201–4212CrossRefGoogle Scholar
  50. 50.
    Pienkos PT, Zhang M (2009) Role of pretreatment and conditioning processes on toxicity of lignocellulosic biomass hydrolysates. Cellulose 16:743–762CrossRefGoogle Scholar
  51. 51.
    Jönsson LJ, Martín C (2016) Pretreatment of lignocellulose: formation of inhibitory by-products and strategies for minimizing their effects. Bioresour Technol 199:103–112CrossRefGoogle Scholar
  52. 52.
    García-Aparicio MP, Ballesteros M, Manzanares P, Ballesteros I, González A, Negro MJ (2007) Xylanase contribution to the efficiency of cellulose enzymatic hydrolysis of barley straw. Appl Biochem Biotechnol 137–140:353–365Google Scholar
  53. 53.
    Yu P, Mckinon JJ, Maenz DD, Olkowski AA, Raez VJ, Christensen DA (2003) Enzymic release of reducing sugars from oat hulls by cellulase, as influenced by Aspergillus ferulic acid esterase and Trichoderma xylanase. J Agric Food Chem 51:218–223CrossRefGoogle Scholar
  54. 54.
    Kumar R, Wyman CE (2009 a) Effects of cellulase and xylanase enzymes on the deconstruction of solids from pretreatment of poplar by leading technologies. Biotechnol Prog 25:302–324CrossRefGoogle Scholar
  55. 55.
    Qing Q, Wyman CE (2011) Supplementation with xylanase and ß-xylosidase to reduce xylo-oligomer and xylan inhibition of enzymatic hydrolysis of cellulose and pretreated corn stover. Biotechnol Biofuels 4(1):2–12CrossRefGoogle Scholar
  56. 56.
    Xiao Z, Zhang X, Gregg DJ, Saddler JN (2004) Effects of sugar inhibition on cellulases and beta-glucosidase during enzymatic hydrolysis of softwood substrates. Appl Biochem Biotechnol 113–116:1115–1126CrossRefGoogle Scholar
  57. 57.
    Kumar R, Wyman CE (2009 b) Effect of enzyme supplementation at moderate cellulase loadings on initial glucose and xylose release from corn stover solids pretreated by leading technologies. Biotechnol Bioeng 102:457–467CrossRefGoogle Scholar
  58. 58.
    Wyman CE, Dale BE, Elander RT, Holtzapple M, Ladisch MR, Lee YY (2005) Comparative sugar recovery data from laboratory scale application of leading pretreatment technologies to corn stover. Bioresour Technol 96:2026–2032CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Division of Crop UtilizationICAR-Central Tuber Crops Research InstituteThiruvananthapuramIndia
  2. 2.Section of Extension and Social SciencesICAR-Central Tuber Crops Research InstituteThiruvananthapuramIndia

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