Advertisement

Addition of Soybean Protein Improves Saccharification and Ethanol Production from Hydrothermally Pretreated Sugarcane Bagasse

  • Camila Florencio
  • Alberto C. Badino
  • Cristiane S. FarinasEmail author
Article
  • 69 Downloads

Abstract

The bioconversion yield of ethanol from lignocellulosic feedstocks is negatively affected by the unproductive adsorption of cellulolytic enzymes onto lignin. In this work, soybean protein was used as a lignin-blocking additive, with the aim of improving the production of ethanol from enzymatic hydrolysates of pretreated sugarcane bagasse. Investigation was made of the effects of the type of hydrothermal pretreatment process—steam explosion (SE) or liquid hot water (LHW), loadings of solids and enzymes, and bioreactor type. The addition of soybean protein led to a exceptional 76% increase of glucose released using the LHW pretreated bagasse, after 24 h of reaction, employing a high-solids loading (15%, w/v) and a low enzyme dosage (5 FPU/g dry biomass). A significant improvement was also achieved for industrial-like mixing conditions in a bench-scale stirred tank reactor, increasing the glucose released by 61 and 42% for the LHW and SE processes, respectively. Ethanol production was also positively affected by the presence of soybean protein, with increases of up to 86 and 65% for the LHW and SE hydrolysates, compared to the control experiment. Characterization of the sugarcane bagasse after the adsorption of soybean protein, using Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM), corroborated the higher affinity of the additive for the LHW bagasse. These findings suggest that soybean protein supplementation during enzymatic hydrolysis by commercially available enzymes is an effective strategy for achieving higher saccharification yields from hydrothermally pretreated biomass, hence improving ethanol production.

Keywords

Enzymatic hydrolysis Lignin Unproductive adsorption Lignocellulosic biomass Sugarcane bagasse Soybean protein 

Notes

Acknowledgements

We are grateful to Dr. Eduardo Ximenes and Dr. Michael Ladisch from Purdue University (IN, USA) for their very insightful suggestions for this work.

Funding information

Embrapa, CNPq (Process 401182/2014-2), CAPES, and FAPESP (Processes 2014/19000-3 and 2016/10636-8) (all from Brazil) provided financial support.

References

  1. 1.
    Lynd LR, Liang XY, Biddy MJ, Allee A, Cai H, Foust T, Himmel ME, Laser MS, Wang M, Wyman CE (2017) Cellulosic ethanol: status and innovation. Curr Opin Biotechnol 45:202–211CrossRefGoogle Scholar
  2. 2.
    Kumar R, Wyman CE (2009) Access of cellulase to cellulose and lignin for poplar solids produced by leading pretreatment technologies. Biotechnol Prog 25:807–819CrossRefGoogle Scholar
  3. 3.
    Lu X, Wang C, Li X, Zhao J (2017) Temperature and pH influence adsorption of cellobiohydrolase onto lignin by changing the protein properties. Bioresour Technol 245:819–825CrossRefGoogle Scholar
  4. 4.
    Tang Y, Chandra RP, Sokhansanj S, Saddler JN (2018) Influence of steam explosion processes on the durability and enzymatic digestibility of wood pellets. Fuel 211:87–94CrossRefGoogle Scholar
  5. 5.
    Ko JK, Kim Y, Ximenes E, Ladisch MR (2015) Effect of liquid hot water pretreatment severity on properties of hardwood lignin and enzymatic hydrolysis of cellulose. Biotechnol Bioeng 112:252–262CrossRefGoogle Scholar
  6. 6.
    Ruiz HA, Rodriguez-Jasso RM, Fernandes BD, Vicente AA, Teixeira JA (2013) Hydrothermal processing, as an alternative for upgrading agriculture residues and marine biomass according to the biorefinery concept: a review. Renew Sustain Energy Rev 21:35–51CrossRefGoogle Scholar
  7. 7.
    Tekin K, Karagoz S, Bektas S (2014) A review of hydrothermal biomass processing. Renew Sustain Energy Rev 40:673–687CrossRefGoogle Scholar
  8. 8.
    Mosier N, Hendrickson R, Ho N, Sedlak M, Ladisch MR (2005) Optimization of pH controlled liquid hot water pretreatment of corn stover. Bioresour Technol 96:1986–1993CrossRefGoogle Scholar
  9. 9.
    Garrote G, Dominguez H, Parajo JC (1999) Hydrothermal processing of lignocellulosic materials. Holz Als Roh-Und Werkstoff 57:191–202CrossRefGoogle Scholar
  10. 10.
    Arenas-Cardenas P, Lopez-Lopez A, Moeller-Chavez GE, Leon-Becerril E (2017) Current pretreatments of lignocellulosic residues in the production of bioethanol. Waste Biomass Valoriz 8:161–181CrossRefGoogle Scholar
  11. 11.
    Kim Y, Kreke T, Hendrickson R, Parenti J, Ladisch MR (2013) Fractionation of cellulase and fermentation inhibitors from steam pretreated mixed hardwood. Bioresour Technol 135:30–38CrossRefGoogle Scholar
  12. 12.
    Ximenes E, Kim Y, Mosier N, Dien B, Ladisch M (2010) Inhibition of cellulases by phenols. Enzym Microb Technol 46:170–176CrossRefGoogle Scholar
  13. 13.
    Ximenes E, Kim Y, Mosier N, Dien B, Ladisch M (2011) Deactivation of cellulases by phenols. Enzym Microb Technol 48:54–60CrossRefGoogle Scholar
  14. 14.
    Kim Y, Ximenes E, Mosier NS, Ladisch MR (2011) Soluble inhibitors/deactivators of cellulase enzymes from lignocellulosic biomass. Enzym Microb Technol 48:408–415CrossRefGoogle Scholar
  15. 15.
    Qin L, Li WC, Liu L, Zhu JQ, Li X, Li BZ, Yuan YJ (2016) Inhibition of lignin-derived phenolic compounds to cellulase. Biotechnol Biofuels 9:10CrossRefGoogle Scholar
  16. 16.
    Kim Y, Kreke T, Ko JK, Ladisch MR (2015) Hydrolysis-determining substrate characteristics in liquid hot water pretreated hardwood. Biotechnol Bioeng 112:677–687CrossRefGoogle Scholar
  17. 17.
    Ko JK, Ximenes E, Kim Y, Ladisch MR (2015) Adsorption of enzyme onto lignins of liquid hot water pretreated hardwoods. Biotechnol Bioeng 112:447–456CrossRefGoogle Scholar
  18. 18.
    Eriksson T, Borjesson J, Tjerneld F (2002) Mechanism of surfactant effect in enzymatic hydrolysis of lignocellulose. Enzym Microb Technol 31:353–364CrossRefGoogle Scholar
  19. 19.
    Lee D, Yu AHC, Saddler JN (1995) Evaluation of cellulase recycling strategies for the hydrolysis of lignocellulosic substrates. Biotechnol Bioeng 45:328–336CrossRefGoogle Scholar
  20. 20.
    Yang B, Wyman CE (2006) Lignin blockers and uses thereof. US PatentGoogle Scholar
  21. 21.
    Yang B, Wyman CE (2006) BSA treatment to enhance enzymatic hydrolysis of cellulose in lignin containing substrates. Biotechnol Bioeng 94:611–617CrossRefGoogle Scholar
  22. 22.
    Kumar R, Wyman CE (2009) Effect of additives on the digestibility of corn Stover solids following pretreatment by leading technologies. Biotechnol Bioeng 102:1544–1557CrossRefGoogle Scholar
  23. 23.
    Florencio C, Badino AC, Farinas CS (2016) Soybean protein as a cost-effective lignin-blocking additive for the saccharification of sugarcane bagasse. Bioresour Technol 221:172–180CrossRefGoogle Scholar
  24. 24.
    Cannella D, Jorgensen H (2014) Do new cellulolytic enzyme preparations affect the industrial strategies for high solids lignocellulosic ethanol production? Biotechnol Bioeng 111:59–68CrossRefGoogle Scholar
  25. 25.
    Jin WX, Chen L, Hu M, Sun D, Li A, Li Y, Hu Z, Zhou SG, Tu YY, Xia T, Wang YT, Xie GS, Li YB, Bai BW, Peng LC (2016) Tween-80 is effective for enhancing steam-exploded biomass enzymatic saccharification and ethanol production by specifically lessening cellulase absorption with lignin in common reed. Appl Energy 175:82–90CrossRefGoogle Scholar
  26. 26.
    Brondi MG, Vasconcellos VM, Giordano RC, Farinas CS (2018) Alternative low-cost additives to improve the saccharification of lignocellulosic biomass. Appl Biochem Biotechnol.  https://doi.org/10.1007/s12010-018-2834-z
  27. 27.
    Carrasco C, Baudel HM, Sendelius J, Modig T, Roslander C, Galbe M, Hahn-Hagerdal B, Zacchi G, Liden G (2010) SO2-catalyzed steam pretreatment and fermentation of enzymatically hydrolyzed sugarcane bagasse. Enzym Microb Technol 46:64–73CrossRefGoogle Scholar
  28. 28.
    Cunha FM, Badino AC, Farinas CS (2017) Effect of a novel method for in-house cellulase production on 2G ethanol yields. Biocatal Agric Biotechnol 9:224–229CrossRefGoogle Scholar
  29. 29.
    Rocha GJM, Silva VFN, Martin C, Goncalves AR, Nascimento VM, Souto-Maior AM (2013) Effect of xylan and lignin removal by hydrothermal pretreatment on enzymatic conversion of sugarcane bagasse cellulose for second generation ethanol production. Sugar Tech 15:390–398CrossRefGoogle Scholar
  30. 30.
    Gouveia ER, do Nascimento RT, Souto-Maior AM, Moraes Rocha GJ (2009) Validation of methodology for the chemical characterization of sugar cane bagasse. Quim Nova 32:1500–1503Google Scholar
  31. 31.
    Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428CrossRefGoogle Scholar
  32. 32.
    Squinca P, Badino AC, Farinas CS (2018) A closed-loop strategy for endoglucanase production using sugarcane bagasse liquefied by a home-made enzymatic cocktail. Bioresour Technol 249:976–982CrossRefGoogle Scholar
  33. 33.
    Ghose TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268Google Scholar
  34. 34.
    Bailey MJ, Poutanen K (1989) Production of xylanolytic enzymes by strains of Aspergillus. Appl Microbiol Biotechnol 30:5–10CrossRefGoogle Scholar
  35. 35.
    Florencio C, Cunha FM, Badino AC, Farinas CS (2015) Validation of a novel sequential cultivation method for the production of enzymatic cocktails from Trichoderma strains. Appl Biochem Biotechnol 175:1389–1402CrossRefGoogle Scholar
  36. 36.
    Pereira SC, Maehara L, Machado CMM, Farinas CS (2016) Physical-chemical-morphological characterization of the whole sugarcane lignocellulosic biomass used for 2G ethanol production by spectroscopy and microscopy techniques. Renew Energy 87:607–617CrossRefGoogle Scholar
  37. 37.
    Sonego JLS, Lemos DA, Rodriguez GY, Cruz AJG, Badino AC (2014) Extractive batch fermentation with CO2 stripping for ethanol production in a bubble column bioreactor: experimental and modeling. Energy Fuel 28:7552–7559CrossRefGoogle Scholar
  38. 38.
    Bondancia TJ, Mattoso LHC, Marconcini JM, Farinas CS (2017) A new approach to obtain cellulose nanocrystals and ethanol from Eucalyptus cellulose pulp via the biochemical pathway. Biotechnol Prog 33:1085–1095CrossRefGoogle Scholar
  39. 39.
    Correa LJ, Badino AC, Cruz AJG (2016) Power consumption evaluation of different fed-batch strategies for enzymatic hydrolysis of sugarcane bagasse. Bioprocess Biosyst Eng 39:825–833CrossRefGoogle Scholar
  40. 40.
    Modenbach AA, Nokes SE (2013) Enzymatic hydrolysis of biomass at high-solids loadings—a review. Biomass Bioenergy 56:526–544CrossRefGoogle Scholar
  41. 41.
    Du J, Li Y, Zhang H, Zheng H, Huang H (2014) Factors to decrease the cellulose conversion of enzymatic hydrolysis of lignocellulose at high solid concentrations. Cellulose 21:2409–2417CrossRefGoogle Scholar
  42. 42.
    Wei WQ, Wu SB (2017) Enhanced enzymatic hydrolysis of eucalyptus by synergy of zinc chloride hydrate pretreatment and bovine serum albumin. Bioresour Technol 245:289–295CrossRefGoogle Scholar
  43. 43.
    Agrawal R, Satlewal A, Kapoor M, Mondal S, Basu B (2017) Investigating the enzyme-lignin binding with surfactants for improved saccharification of pilot scale pretreated wheat straw. Bioresour Technol 224:411–418CrossRefGoogle Scholar
  44. 44.
    Zhang HD, Ye GY, Wei YT, Li X, Zhang AP, Xie J (2017) Enhanced enzymatic hydrolysis of sugarcane bagasse with ferric chloride pretreatment and surfactant. Bioresour Technol 229:96–103CrossRefGoogle Scholar
  45. 45.
    Buffo MM, Correa LJ, Esperanca MN, Cruz AJG, Farinas CS, Badino AC (2016) Influence of dual-impeller type and configuration on oxygen transfer,. Power consumption, and shear rate in a stirred tank bioreactor. Biochem Eng J 114:133–142CrossRefGoogle Scholar
  46. 46.
    Correa LJ, Badino AC, Goncalves Cruz AJ (2016) Mixing design for enzymatic hydrolysis of sugarcane bagasse: methodology for selection of impeller configuration. Bioprocess Biosyst Eng 39:285–294CrossRefGoogle Scholar
  47. 47.
    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
  48. 48.
    Carvalheiro F, Duarte LC, Girio FM (2008) Hemicellulose biorefineries: a review on biomass pretreatments. J Sci Ind Res 67:849–864Google Scholar
  49. 49.
    Bhagia S, Kumar R, Wyman CE (2017) Effects of dilute acid and flowthrough pretreatments and BSA supplementation on enzymatic deconstruction of poplar by cellulase and xylanase. Carbohydr Polym 157:1940–1948CrossRefGoogle Scholar
  50. 50.
    Brethauer S, Studer MH, Yang B, Wyman CE (2011) The effect of bovine serum albumin on batch and continuous enzymatic cellulose hydrolysis mixed by stirring or shaking. Bioresour Technol 102:6295–6298CrossRefGoogle Scholar
  51. 51.
    Hsieh C-w C, Cannella D, Jorgensen H, Felby C, Thygesen LG (2015) Cellobiohydrolase and endoglucanase respond differently to surfactants during the hydrolysis of cellulose. Biotechnol Biofuels 8:52CrossRefGoogle Scholar
  52. 52.
    Yang M, Zhang JH, Kuittinen S, Vepsalainen J, Soininen P, Keinanen M, Pappinen A (2015) Enhanced sugar production from pretreated barley straw by additive xylanase and surfactants in enzymatic hydrolysis for acetone-butanol-ethanol fermentation. Bioresour Technol 189:131–137CrossRefGoogle Scholar
  53. 53.
    Jackson M, Mantsch HH (1995) The use and misuse of FTIR spectroscopy in the determination of protein structure. Crit Rev Biochem Mol Biol 30:95–120CrossRefGoogle Scholar
  54. 54.
    Rodriguez-Zuniga UF, Neto VB, Couri S, Crestana S, Farinas CS (2014) Use of spectroscopic and imaging techniques to evaluate pretreated sugarcane bagasse as a substrate for cellulase production under solid-state fermentation. Appl Biochem Biotechnol 172:2348–2362CrossRefGoogle Scholar
  55. 55.
    Kristensen JB, Thygesen LG, Felby C, Jorgensen H, Elder T (2008) Cell-wall structural changes in wheat straw pretreated for bioethanol production. Biotechnol Biofuels 1:5CrossRefGoogle Scholar
  56. 56.
    Schwanninger M, Rodrigues JC, Pereira H, Hinterstoisser B (2004) Effects of short-time vibratory ball milling on the shape of FT-IR spectra of wood and cellulose. Vib Spectrosc 36:23–40CrossRefGoogle Scholar
  57. 57.
    Qi GX, Peng F, Xiong L, Lin XQ, Huang C, Li HL, Chen XF, Chen XD (2017) Extraction and characterization of wax from sugarcane bagasse and the enzymatic hydrolysis of dewaxed sugarcane bagasse. Prep Biochem Biotechnol 47:276–281CrossRefGoogle Scholar
  58. 58.
    Camargo LA, Pereira SC, Correa AC, Farinas CS, Marconcini JM, Mattoso LHC (2016) Feasibility of manufacturing cellulose nanocrystals from the solid residues of second-generation ethanol production from sugarcane bagasse. Bioenergy Res 9:894–906CrossRefGoogle Scholar
  59. 59.
    Lu XQ, Zheng XJ, Li XZ, Zhao J (2016) Adsorption and mechanism of cellulase enzymes onto lignin isolated from corn stover pretreated with liquid hot water. Biotechnol Biofuels 9:118CrossRefGoogle Scholar
  60. 60.
    Novaes Reis Corrales RC, Teixeira Mendes FM, Perrone CC, Sant'Anna C, de Souza W, Abud Y, da Silva Bon EP, Ferreira-Leitao V (2012) Structural evaluation of sugar cane bagasse steam pretreated in the presence of CO2 and SO2. Biotechnol Biofuels 5:36.  https://doi.org/10.1186/1754-6834-5-36
  61. 61.
    Xu Z, Wang Q, Jiang Z, Yang X-x, Ji Y (2007) Enzymatic hydrolysis of pretreated soybean straw. Biomass Bioenergy 31:162–167CrossRefGoogle Scholar
  62. 62.
    Crepin L, Truong NM, Bloem A, Sanchez I, Dequin S, Camarasa C (2017) Management of multiple nitrogen sources during wine fermentation by Saccharomyces cerevisiae. Appl Environ Microbiol 83(5):e02617-16Google Scholar
  63. 63.
    Lekkas C, Stewart GG, Hill AE, Taidi B, Hodgson J (2012) Elucidation of the role of nitrogenous wort components in yeast fermentation. J Inst Brew 113:3–8CrossRefGoogle Scholar
  64. 64.
    Klein-Marcuschamer D, Oleskowicz-Popiel P, Simmons BA, Blanch HW (2012) The challenge of enzyme cost in the production of lignocellulosic biofuels. Biotechnol Bioeng 109:1083–1087CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Embrapa InstrumentaçãoSão CarlosBrazil
  2. 2.Graduate Program of Chemical EngineeringFederal University of São CarlosSão CarlosBrazil

Personalised recommendations