Skip to main content
SpringerLink
Log in

The effect of agitation speed, enzyme loading and substrate concentration on enzymatic hydrolysis of cellulose from brewer’s spent grain

  • Published:
Cellulose Aims and scope Submit manuscript

Abstract

Brewer’s spent grain components (cellulose, hemicellulose and lignin) were fractionated in a two-step chemical pretreatment process using dilute sulfuric acid and sodium hydroxide solutions. The cellulose pulp produced was hydrolyzed with a cellulolytic complex, Celluclast 1.5 L, at 45 °C to convert the cellulose into glucose. Several conditions were examined: agitation speed (100, 150 and 200 rpm), enzyme loading (5, 25 and 45 FPU/g substrate), and substrate concentration (2, 5 and 8% w/v), according to a 23 full factorial design aiming to maximize the glucose yield. The obtained results were interpreted by analysis of variance and response surface methodology. The optimal conditions for enzymatic hydrolysis of brewer’s spent grain were identified as 100 rpm, 45 FPU/g and 2% w/v substrate. Under these conditions, a glucose yield of 93.1% and a cellulose conversion (into glucose and cellobiose) of 99.4% was achieved. The easiness of glucose release from BSG makes this substrate a raw material with great potential to be used in bioconversion processes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Berlin A, Gilkes N, Kilburn D, Bura R, Markov A, Skomarovsky A, Okunev O, Gusakov A, Maximenko V, Gregg D, Sinitsyn A, Saddler J (2005) Evaluation of novel fungal cellulose preparations for ability to hydrolyze softwood substrates—evidence for the role of accessory enzymes. Enzyme Microb Technol 37:175–184

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Cao Y, Tan H (2002) Effects of cellulase on the modification of cellulose. Carbohydr Res 337:1291–1296

    Article  CAS  Google Scholar 

  • Chen M, Xia L, Xue P (2007) Enzymatic hydrolysis of corncob and ethanol production from cellulosic hydrolysate. Int Biodeter Biodegr 59:85–89

    Article  CAS  Google Scholar 

  • Converse AO, Ooshima H, Burns DS (1990) Kinetics of enzymatic hydrolysis of lignocellulosic materials based on surface area of cellulose accessible to enzyme and enzyme adsorption on lignin and cellulose. Appl Biochem Biotechnol 24/25:67–73

    Article  Google Scholar 

  • Eklund R, Galbe M, Zacchi G (1990) Optimization of temperature and enzyme concentration in the enzymatic saccharification of steam-pretreated willow. Enzyme Microb Technol 12:225–228

    Article  CAS  Google Scholar 

  • Frennesson I, Trägardh G, Hahn-Hägerdal B (1985) An ultrafiltration membrane reactor for obtaining experimental reaction rates at defined concentrations of inhibiting sugars during enzymatic saccharification of alkali-pretreated sallow: formulation of a simple empirical rate equation. Biotechnol Bioeng 27:1328–1334

    Article  CAS  Google Scholar 

  • Gan Q, Allen SJ, Taylor G (2002) Design and operation of an integrated membrane reactor for enzymatic cellulose hydrolysis. Biochem Eng J 12:223–229

    Article  CAS  Google Scholar 

  • Gan Q, Allen SJ, Taylor G (2003) Kinetic dynamics in heterogeneous enzymatic hydrolysis of cellulose: an overview, an experimental study and mathematical modeling. Process Biochem 38:1003–1018

    Article  CAS  Google Scholar 

  • Gharpuray MM, Lee Y-H, Fan LT (1983) Structural modification of lignocellulosics by pretreatments to enhance enzymatic hydrolysis. Biotechnol Bioeng 25:157–172

    Article  CAS  Google Scholar 

  • Ingesson H, Zacchi G, Yang B, Esteghlalian AR, Saddler JN (2001) The effect of shaking regime on the rate and extent of enzymatic hydrolysis of cellulose. J Biotechnol 88:177–182

    Article  CAS  Google Scholar 

  • Kaur PP, Arneja JS, Singh J (1998) Enzymic hydrolysis of rice straw by crude cellulose from Trichoderma reesei. Bioresour Technol 66:267–269

    Article  CAS  Google Scholar 

  • Kaya F, Heitmann JA, Joye TW (2000) Influence of lignin and its degradation products on enzymatic hydrolysis of xylan. J Biotechnol 80:241–247

    Article  CAS  Google Scholar 

  • Liao W, Wen Z, Hurley S, Liu Y, Liu C, Chen S (2005) Effects of hemicellulose and lignin on enzymatic hydrolysis of cellulose from dairy manure. Appl Biochem Biotechnol 121/124:1017–1029

    Article  Google Scholar 

  • Mandels M, Andreotti R, Roche C (1976) Measurement of saccharifying cellulase. Biotechnol Bioeng Symp 6:17–34

    Google Scholar 

  • Manonmani HK, Sreekantiah KR (1987) Saccharification of sugarcane bagasse with enzymes from Aspergillus ustus and Trichoderma viride. Enzyme Microb Technol 9:484–488

    Article  CAS  Google Scholar 

  • Mansfield SD, Mooney C, Saddler JN (1999) Substrate and enzyme characteristics that limit cellulose hydrolysis. Biotechnol Prog 15:804–816

    Article  CAS  Google Scholar 

  • Martín C, Galbe M, Nilvebrant N-O, Jönsson LJ (2002) Comparison of the fermentability of enzymatic hydrolyzates of sugarcane bagasse pretreated by steam explosion using different impregnating agents. Appl Biochem Biotechnol 98/100:699–716

    Article  Google Scholar 

  • McMillan JD (1994) Pretreatment of lignocellulosic biomass. In: Himmel ME, Baker LO, Overend RP (eds) Conversion of hemicellulose hydrolyzates to ethanol, American Chemical Society Symposium, Washington, pp 292–324

    Google Scholar 

  • Miller GM (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Moldes AB, Alonso JL, Parajó JC (1999) Cogeneration of cellobiose and glucose from pretreated wood and bioconversion to lactic acid: a kinetic study. J Biosci Bioeng 87:787–792

    Article  CAS  Google Scholar 

  • Moriyama S, Saida T (1986) Continuous pretreatment and enzymatic saccharification of lignocellulosics. In: Young RA, Rowell RM (eds) Cellulose: structure, modification and hydrolysis, Wiley Interscience, New York, pp 323–335

    Google Scholar 

  • Mukataka S, Tada M, Takahashi J (1983) Effects of agitation on enzymatic hydrolysis of cellulose in a stirred tank reactor. J Ferment Technol 61:615–621

    CAS  Google Scholar 

  • Mussatto SI, Rocha GJM, Roberto IC Hydrogen peroxide bleaching of cellulose pulps obtained from brewer’s spent grain. Cellulose. doi:10.100/s105-008-919-4 (in press)

  • Mussatto SI, Dragone G, Roberto IC (2006a) Brewer’s spent grain: generation, characteristics and potential applications. J Cereal Sci 43:1–14

    Article  CAS  Google Scholar 

  • Mussatto SI, Dragone G, Rocha GJM, Roberto IC (2006b) Optimum operating conditions for brewer’s spent grain soda pulping. Carbohydr Polym 64:22–28

    Article  CAS  Google Scholar 

  • Mussatto SI, Roberto IC (2005) Acid hydrolysis and fermentation of brewer’s spent grain to produce xylitol. J Sci Food Agric 85:2453–2460

    Article  CAS  Google Scholar 

  • Mussatto SI, Roberto IC (2006) Chemical characterization and liberation of pentose sugars from brewer’s spent grain. J Chem Technol Biotechnol 81:268–274

    Article  CAS  Google Scholar 

  • Pan X, Xie D, Gilkes N, Gregg DJ, Saddler JN (2005) Strategies to enhance the enzymatic hydrolysis of pretreated softwood with high residual lignin content. Appl Biochem Biotechnol 121/124:1069–1079

    Article  Google Scholar 

  • Roberto IC, Mussatto SI, Rodrigues RCLB (2003) Dilute-acid hydrolysis for optimization of xylose recovery from rice straw in a semi-pilot reactor. Ind Crop Prod 17:171–176

    Article  CAS  Google Scholar 

  • Sattler W, Esterbauer H, Glatter O, Steiner W (1989) The effect of enzyme concentration on the rate of the hydrolysis of cellulose. Biotechnol Bioeng 33:1221–1234

    Article  CAS  Google Scholar 

  • Sharma A, Khare SK, Gupta MN (2001) Hydrolysis of rice hull by crosslinked Aspergillus niger cellulose. Bioresour Technol 78:281–284

    Article  CAS  Google Scholar 

  • Tengborg C, Galbe M, Zacchi G (2001) Influence of enzyme loading and physical parameters on the enzymatic hydrolysis of steam-pretreated softwood. Biotechnol Prog 17:110–117

    Article  CAS  Google Scholar 

  • Varga E, Szengyel Z, Réczey K (2002) Chemical pretreatments of corn stover for enhancing enzymatic digestibility. Appl Biochem Biotechnol 98/100:73–87

    Article  Google Scholar 

  • Vlasenko EY, Ding H, Labavitch JM, Shoemaker SP (1997) Enzymatic hydrolysis of pretreated rice straw. Bioresour Technol 59:109–119

    Article  CAS  Google Scholar 

  • Wen Z, Liao W, Chen S (2004) Hydrolysis of animal manure lignocellulosics for reducing sugar production. Bioresour Technol 91:31–39

    Article  CAS  Google Scholar 

  • Wright JD (1988) Ethanol from biomass by enzymatic hydrolysis. Chem Eng Prog 89:62–74

    Google Scholar 

  • Wu MM, Chang K, Gregg DJ, Boussaid A, Blatson RP, Saddler JN (1999) Optimization of steam explosion to enhance hemicellulose recovery and enzymatic hydrolysis of cellulose in softwoods. Appl Biochem Biotechnol 77/79:47–54

    Article  Google Scholar 

  • Yáñez R, Alonso JL, Parajó JC (2006) Enzymatic saccharification of hydrogen-peroxide-treated solids from hydrothermal processing of rice husks. Process Biochem 41:1244–1252

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by Capes (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo), Brazil. The authors also acknowledge Novozymes, who supplied the enzyme preparation used in this study.

Author information

Authors and Affiliations

  1. Departamento de Biotecnologia, Escola de Engenharia de Lorena, Universidade de São Paulo, Lorena, SP, Brazil

    Solange I. Mussatto, Giuliano Dragone, Marcela Fernandes, Adriane M. F. Milagres & Inês C. Roberto

  2. Departamento de Engenharia Biológica, Universidade do Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    Solange I. Mussatto

Authors
  1. Solange I. Mussatto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Giuliano Dragone
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marcela Fernandes
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Adriane M. F. Milagres
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Inês C. Roberto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Solange I. Mussatto.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mussatto, S.I., Dragone, G., Fernandes, M. et al. The effect of agitation speed, enzyme loading and substrate concentration on enzymatic hydrolysis of cellulose from brewer’s spent grain. Cellulose 15, 711–721 (2008). https://doi.org/10.1007/s10570-008-9215-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10570-008-9215-7

Keywords

Search

Navigation