, Volume 25, Issue 4, pp 2487–2504 | Cite as

Statistical modelling and optimization of alkaline peroxide oxidation pretreatment process on rice husk cellulosic biomass to enhance enzymatic convertibility and fermentation to ethanol

  • Augustine Omoniyi Ayeni
  • Michael Olawale Daramola
  • Patrick T. Sekoai
  • Opeyemi Adeeyo
  • Musa Joel Garba
  • Ayotunde A. Awosusi
Original Paper


The complex and ordered arrangements of the lignocellulosic materials make them recalcitrant for their conversions to ethanol. Pretreatment is a crucial step in overcoming these hindrances. In this study, a 23-full factorial design of experiments optimization technique was applied on the alkaline peroxide oxidation pretreatments of rice husks biomass. The low–high levels of the influencing variables on pretreatments were; temperature (100–120 °C), time (1–2 h), % (v/v)H2O2 concentration (1–3%). Under the prevailing pretreatments, the optimum conditions were predicted and validated to be 109 °C, 2 h, and 1.38% H2O2 which yielded 56% (w/w) cellulose content, 55% (w/w) hemicellulose solubilization, and 48% (w/w) lignin removal. At the established optimum pretreatment conditions, and considering variations in biomass and enzymes loadings, maximum reducing sugars production was 205 mg/g dry biomass at different enzymatic hydrolysis conditions of 3% biomass loading, hydrolysis temperature of 45 °C, hydrolysis time of 24 h, and 35 FPU/g cellulose enzyme loading. The highest cellulose conversion of 33% yielded 24 g/L ethanol at the end of the first day of saccharification and fermentation. Physical, structural, and morphological investigations on raw and treated materials using tools such as stereomicroscopy, scanning electron microscopy, and fourier transform infrared spectroscopy further revealed the effectiveness of chosen method on rice husks biomass.


Full factorial design Alkaline peroxide oxidation Enzymatic convertibility Scanning electron microscopy Cellulosic biomass Ethanol 


  1. Agnemo R, Gellerstedt G (1979) Reactions of lignin with alkaline hydrogen peroxide. Part II. Factors influencing the decomposition of phenolic structures. Acta Chem Scand B33:337–342CrossRefGoogle Scholar
  2. Akerholm M, Hinterstoisser B, Salmen L (2004) Characterization of the crystalline structure of cellulose using static and dynamic FT-IR spectroscopy. Carbohydr Res 339:569–578CrossRefGoogle Scholar
  3. Andrić P, Meyer AS, Jensen PA, Dam-Johansen K (2010) Reactor design for minimizing product inhibition during enzymatic lignocellulosic hydrolysis II. Quantification of inhibition and suitability of membrane reactors. Biotechnol Adv 28:407–425CrossRefGoogle Scholar
  4. Asadieraghi M, Ashri WM, Daud W (2014) Characterization of lignocellulosic biomass thermal degradation and physiochemical structure: effect of demineralization by diverse acid solutions. Energy Convers Manag 82:71–82CrossRefGoogle Scholar
  5. ASTM D2015. Standard test method for gross caloric value of coal and coke by adiabatic bomb calorimeter.
  6. Awosusi AA, Ayeni A, Adeleke R, Daramola MO (2017a) Effect of water of crystallization on the dissolution efficiency of molten zinc chloride hydrate salts during the pretraetment of corncob biomass. J Chem Tech Biotechnol 92:2468–2476CrossRefGoogle Scholar
  7. Awosusi AA, Ayeni A, Adeleke R, Daramola MO (2017b) Biocompositional and thermodecompositional analysis of South African agro-waste corncob and husk towards production of biocommodities. Asia Pac J Chem Eng 12:960–968CrossRefGoogle Scholar
  8. Ayeni AO, Daramola MO (2017) Lignocellulosic biomass waste beneficiation: evaluation of oxidative and non-oxidative pretreatment methodologies of South African corn cob. J Environ Chem Eng 5:1771–1779CrossRefGoogle Scholar
  9. Ayeni AO, Banerjee S, Omoleye JA, Hymore FK, Giri BS, Deshmukh SC, Pandey RA, Mudliar SN (2013a) Optimization of pretreatment conditions using full factorial design and enzymatic convertibility of shea tree sawdust. Biomass Bioenergy 48:130–138CrossRefGoogle Scholar
  10. Ayeni AO, Hymore FK, Mudliar SN, Deskmukh SC, Satpute DB, Omoleye JA, Pandey RA (2013b) Hydrogen peroxide and lime based oxidative pretreatment of wood waste to enhance enzymatic hydrolysis for a biorefinery: process parameters optimization using response surface methodology. Fuel 106:187–194CrossRefGoogle Scholar
  11. Ayeni AO, Omoleye JA, Mudliar SN, Hymore FK, Pandey RA (2014) Utilization of lignocellulosic waste for ethanol production: enzymatic digestibility and fermentation of pretreated shea tree sawdust. Korean J Chem Eng 31:1180–1186CrossRefGoogle Scholar
  12. Ayeni AO, Omoleye JA, Hymore FK, Pandey RA (2016a) Effective alkaline peroxide oxidation pretreatment of shea tree sawdust for the production of biofuels: kinetics of delignification and enzymatic conversion to sugar and subsequent production of ethanol by fermentation using Saccharomyces cerevisiae. Braz J Chem Eng 33:33–45CrossRefGoogle Scholar
  13. Ayeni AO, Ogu R, Awosusi AA, Daramola MO (2016b) Alkaline peroxide oxidation pretreatment of corn cob and rice husks for bioconversion into bio-commodities: Part A-Enzymatic converstibility of pretreated rice husks to reducing sugar. In: 24th European biomass conference and exhibition, pp 1225–1232. Amsterdam, NetherlandsGoogle Scholar
  14. Bailey CW, Dence CW (1969) Reactions of alkaline hydrogen peroxide with softwood lignin model compounds. Tappi J 52:491–500Google Scholar
  15. Banerjee S, Sen R, Pandey R, Chakrabarti T, Satpute D, Giri B, Mudliar SN (2009) Evaluation of wet air oxidation as a pretreatment strategy for bioethanol production from rice husk and process optimization. Biomass Bioenergy 33:1680–1686CrossRefGoogle Scholar
  16. Bartos C, Kukovecz Á, Ambrus R, Farkas G, Radacsi N, Szabó-Révész P (2015) Comparison of static and dynamic sonication as process intensification for particle size reduction using a factorial design. Chem Eng Process 87:26–34CrossRefGoogle Scholar
  17. Basu P (2010) Biomass gasification and pyrolysis: practical design and theory. Elsevier, OxfordGoogle Scholar
  18. Bennet C (1971) Spectrophotometric acid dichromate method for the determination of ethyl alcohol. Am J Med Technol 37:217–220Google Scholar
  19. Bian J, Peng F, Peng X-P, Peng P, Xu F, Sun R-C (2012) Acetic acid enhanced purification of crude cellulose from sugarcane bagasse: structural and morphological characterization. Bioresources 7:4626–4639CrossRefGoogle Scholar
  20. Binder JB, Raines RT (2010) Fermentable sugars by chemical hydrolysis of biomass. Proc Natl Acad Sci USA 107:4516–4521CrossRefGoogle Scholar
  21. Blasi CD, Signorelli G, Russo CD, Rea G (1999) Product distribution from pyrolysis of wood and agricultural residues. Ind Eng Chem Res 38:2216–2224CrossRefGoogle Scholar
  22. Boie W (1957) Vom Brennstoff zum Rauchgas, in: Feuerungstechnisches Rechnen mit Brennstoffkenngr¨oßen und seine Vereinfachung mitMitteln der Statistik, Teubner, LeipzigGoogle Scholar
  23. Buzala K, Przybysz P, Rosicka-Kaczmarek J, Kalinowska H (2015) Production of glucose-rich enzymatic hydrolysates from cellulosic pulps. Cellulose 22:663–674CrossRefGoogle Scholar
  24. Chang VS, Nagwani M, Holtzapple MT (1998) Lime pretreatment of crop residues bagasse and wheat straw. Appl Biochem Biotechnol 74:135–159CrossRefGoogle Scholar
  25. Chundawat SPS, Donohoe BS, Sousa LD, Elder T, Agarwal UP, Lu F, Ralph J, Himmel ME, Balan V, Dale BE (2011) Multi-scale visualization and characterization of lignocellulosic plant cell wall deconstruction during thermochemical pretreatment. Energy Environ Sci 4:973–984CrossRefGoogle Scholar
  26. Ciesielski PN, Wang W, Chen X, Vinzant TB, Tucker MP, Decker SR, Himmel ME, Johnson DK, Donohoe BS (2014) Effect of mechanical disruption on the effectiveness of three reactors used for dilute acid pretreatment of corn stover Part 2: morphological and structural substrate analysis. Biotechnol Biofuels 7:47CrossRefGoogle Scholar
  27. Dagnino EP, Chamorro ER, Romano SD, Felissia FE, Area MC (2013) Optimization of the acid pretreatment of rice hulls to obtain fermentable sugars for bioethanol production. Ind Crops Prod 42:363–368CrossRefGoogle Scholar
  28. Demirbas A (1997) Calculation of high heating values of biomass fuels. Fuel 76:431–434CrossRefGoogle Scholar
  29. Donohoe BS, Decker SR, Tucker MP, Himmel ME, Vinzant TB (2008) Visualizing lignin coalescence and migration through maize cell walls following thermochemical pretreatment. Biotechnol Bioeng 101:913–925CrossRefGoogle Scholar
  30. Dowe N, McMillan J (2008) SSF experimental protocols: Lignocellulosic biomass hydrolysis and fermentation LAP. NREL/TP-510-42630. Contract No.: DE-AC36-99-G010337Google Scholar
  31. El-Sayed SA, Mostafa ME (2015) Kinetic parameters determination of biomass pyrolysis fuels using TGA and DTA techniques. Waste Biomass Valoris 6:401–415CrossRefGoogle Scholar
  32. Fannie PE, Okafor J, Roberson C (1998) Determination of insoluble solids of pretreated biomass material. LAP-18. Version 09-23-1998Google Scholar
  33. Forney LJ, Reddy CA, Tien M, Aust SD (1982) The involvement of hydroxyl radical derived from hydrogen peroxide in lignin degradation by the white rot fungus phanerochaete chrysosporium. J Biol Chem 257:11455–11462Google Scholar
  34. García R, Pizarro C, Lavín AG, Bueno JL (2014) Spanish biofuels heating value estimation. Part 1: Ultimate analysis data. Fuel 117:1130–1138CrossRefGoogle Scholar
  35. Gould JM (1985) Studies on the mechanism of alkaline peroxide delignification of agricultural residues. Biotechnol Bioeng 27:225–231CrossRefGoogle Scholar
  36. Hsu T, Guo G, Chen W, Hwang W (2010) Effect of dilute acid pretreatment of rice straw on structural properties and enzymatic hydrolysis. Bioresour Technol 101:4907–4913CrossRefGoogle Scholar
  37. Jenkins BM, Ebeling JM (1985) Correlations of physical and chemical properties of terrestrial biomass with conversion. In: Proceedings of 1985 symposium energy from biomass and waste IX IGT, p 271, CaliforniaGoogle Scholar
  38. Jeya M, Zhang Y, Kim I, Lee J (2009) Enhanced saccharification of alkali-treated rice straw by cellulase from Trametes hirsuta and statistical optimization of hydrolysis conditions by RSM. Bioresour Technol 100:5155–5161CrossRefGoogle Scholar
  39. Jung KW, Kim DH, Kim HW, Shin HS (2011) Optimization of combined (acid + thermal) pretreatment for fermentative hydrogen production from Laminaria japonica using response surface methodology (RSM). Int J Hydrog Energy 36:9626–9631CrossRefGoogle Scholar
  40. Kaiser S, Verza SG, Moraes RC, Pittol V, Peñaloza EMC, Pavei C, Ortega GG (2013) Extraction optimization of polyphenols, oxindole alkaloids and quinovic acid glycosides from cat’s claw bark by Box-Behnken design. Ind Crops Prod 48:153–161CrossRefGoogle Scholar
  41. Kim S, Dale BE (2004) Global potential bioethanol production from wasted crops and crop residues. Biomass Bioenergy 26:361–375CrossRefGoogle Scholar
  42. Kutsuki H, Gold MH (1982) Generation of hydroxyl radical and its involvement in lignin degradation by Phanerochaete chrysosporium. Biochem Biophys Res Cornrnun 109:320–327CrossRefGoogle Scholar
  43. Lachenal D, de Choudens C, Monzie P (1980) Hydrogen peroxide as a delignifying agent. Tappi J 63:119–122Google Scholar
  44. Lee JH, Lim SL, Song YS, Kang SW, Park C, Kim SW (2007) Optimization of culture medium for lactosucrose (4G-β-D-Galactosylsucrosw) production by Sterigmatomyces elviae mutant using statistical analysis. J Microbiol Biotechnol 17:1996–2004Google Scholar
  45. Li S, Xu S, Liu S, Yang C, Lu Q (2004) Fast pyrolysis of biomass in free—fall reactor for hydrogen—rich gas. Fuel Process Technol 85:1201–1211CrossRefGoogle Scholar
  46. Mani T, Murugan P, Abedi J, Mahinpey N (2010) Pyrolysis of wheat straw in a thermogravimetric analyser: effect of particle size and heating rate on devolatilization and estimation of global kinetics. Chem Eng Res Des 88:952–958CrossRefGoogle Scholar
  47. McGinnis GD, Prince SE, Biermann CJ, Lowrimore JF (1984) Wet oxidation of model carbohydrate compounds. Carbohyd Res 128:51–60CrossRefGoogle Scholar
  48. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428CrossRefGoogle Scholar
  49. Nelson ML, O’Connor RT (1964) Relation of certain infrared bands to cellulose crystallinity and crystal lattice type. Part II. A new infrared ratio for estimation of crystallinity in cellulose I and II. J Appl Polym Sci 8:1325–1341CrossRefGoogle Scholar
  50. Nikzad M, Movagharnejad K, Najafpour GD, Talebnia F (2013) Comparative studies of effect of pretreatment of rice husk for enzymatic digestibility and bioethanol production. Int J Eng 26:455–464CrossRefGoogle Scholar
  51. O’Sullivan AC (1997) Cellulose: the structure slowly unravels. Cellulose 4:173–207CrossRefGoogle Scholar
  52. Onoji SE, Iyuke SE, Igbafe AI, Daramola MO (2017) Hevea brasiliensis (rubber seed) oil: modelling and optimization of extraction process parameters using response surface methodology and artificial neural network techniques. Biofuels. Google Scholar
  53. Park S, Baker JO, Himmel ME, Parilla PA, Johnson DK (2010) Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance. Biotechnol Biofuels 3:10CrossRefGoogle Scholar
  54. Permchart W, Kouprianov VI (2004) Emission performance and combustion efficiency of a conical fluidized-bed combustor firing various biomass fuels. Bioresour Technol 92:83–91. CrossRefGoogle Scholar
  55. Saha BC, Cotta MA (2007) Enzymatic saccharification and fermentation of alkaline peroxide pretreated rice hulls to ethanol. Enzyme Microb Technol 41:528–532CrossRefGoogle Scholar
  56. Sarita CR, Filho RM, Costa AC (2009) Lime pretreatment of sugarcane bagasse for bioethanol production. Appl Biochem Biotechnol 153:139–150CrossRefGoogle Scholar
  57. Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D (2008) Determination of Ash in Biomass. LAP. NREL/TP 510-42622Google Scholar
  58. Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D, Crocker D (2012) Determination of structural carbohydrates and lignin in biomass. LAP. NREL/TP 510-42618. Version 08-03-2012Google Scholar
  59. Spiridon I, Teacã C, Bodîrlãu R (2011) Structural changes evidenced by FTIR spectroscopy in cellulosic materials after pretreatment with ionic liquid and enzymatic hydrolysis. Bioresources 6:400–413Google Scholar
  60. Stasolla C, Scott J, Egertsdotter U, Kadla J, O’Malley D, Sederoff R, Zyl L (2003) Analysis of lignin produced by cinnamyl alcohol dehydrogenase-deficient Pinus taeda cultured cells. Plant Physiol Biochem 41:439–445CrossRefGoogle Scholar
  61. Tengborg C, Galbe M, Zacchi G (2001) Influence of enzyme loading and physical parameters on the enzymatic hydrolysis of steam pretreated softwood. Biotechnol Progress 17:110–117CrossRefGoogle Scholar
  62. Tokoh C, Takabe K, Fujita M, Saiki H (1998) Cellulose synthesized by Azetobacter xylinum in the presence of acetyl glucomannan. Cellulose 5:249–261CrossRefGoogle Scholar
  63. Varanasi P, Singh P, Auer M, Adams PD, Simmons BA, Singh S (2013) Survey of renewable chemicals produced from lignocellulosic biomass during ionic liquid pretreatment. Biotechnol Biofuels 6:14CrossRefGoogle Scholar
  64. Viamajala S, McMillan JD, Schell DJ, Elander RT (2009) Rheology of corn stover slurries at high solids concentrations–Effects of saccharification and particle size. Bioresour Technol 100:925–934CrossRefGoogle Scholar
  65. Wang K, Yang H, Wang W, Sun R (2013) Structural evaluation and bioethanol production by simultaneous saccharification and fermentation with biodegraded triploid poplar. Biotechnol Biofuels 6:42CrossRefGoogle Scholar
  66. Wright MM, Daugaard DE, Satrio JA, Brown RC (2010) Techno-economic analysis of biomass fast pyrolysis to transportation fuels. Fuel 89:S2–S10CrossRefGoogle Scholar
  67. Xing Y, Bu L, Sun D, Liu Z, Liu S, Jiang J (2016) Enhancement of high-solid enzymatic hydrolysis and fermentation of furfural residues by addition of Gleditsia saponin. Fuel 177:142–147CrossRefGoogle Scholar
  68. Yang B, Wyman CE (2008) Pretreatment: the key to unlocking low-cost cellulosic ethanol. Biofuels Bioprod Biorefin 2:26–40CrossRefGoogle Scholar
  69. Yin C (2011) Prediction of higher heating values of biomass from proximate and ultimate analyses. Fuel 90:1128–1132CrossRefGoogle Scholar
  70. Zhang Q, Cai W (2008) Enzymatic hydrolysis of alkali-pretreated rice straw by Trichoderma reesei ZM4-F3. Biomass Bioenergy 32:1130–1135CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Chemical Engineering, College of EngineeringCovenant UniversityOtaNigeria
  2. 2.Sustainable Energy and Environment Research Unit, School of Chemical and Metallurgical Engineering, Faculty of Engineering and the Built EnvironmentUniversity of the Witwatersrand (Wits)JohannesburgSouth Africa

Personalised recommendations