Skip to main content
SpringerLink
Log in

A new approach for Chrysoporthe cubensis cellulolytic cocktail production using solid and submerged-state fermentation

  • Original Paper
  • Published:
Brazilian Journal of Chemical Engineering Aims and scope Submit manuscript

Abstract

The lignocellulosic material bioconversion to bioproducts has received significant attention in recent years. Cellulases and hemicellulases catalyze the hydrolysis of lignocellulosic materials into fermentable sugars that are afterward converted to bioproducts by microorganisms. Chrysoporthe cubensis grown under solid-state fermentation (SSF) has produced more effective enzymatic extracts for sugarcane bagasse saccharification than commercial cellulolytic preparations. However, the investigation of new approaches for enzyme production by this fungus is still lacking. In this work, an enzyme cocktail (SSF-SmF-cocktail) was produced by extracting enzymes of C. cubensis grown under SSF using the extract produced by the same fungus under submerged fermentation (SmF). The total cellulase (FPase), carboxymethylcellulase (CMCase), cellobiohydrolase (CBH), β-glucosidase, xylanase, β-xylosidase, β-galactosidase, α-galactosidase, and α-arabinofuranosidase activities were evaluated in crude extracts obtained from C. cubensis cultivation under SSF, SmF, and also in the SSF-SmF-cocktail. The C. cubensis protein profiles cultivated under SSF and SmF were compared by SDS-PAGE. Extract produced by C. cubensis grown under SmF presented proteins with estimated molecular weights of 10.7, 29.3, 38.6, 46.0, and 170.0 kDa, respectively, but not in that produced by this fungus under SSF. When cultivated under SSF, C. cubensis produced an extract with greater protein diversity between 13 and 51 kDa than that obtained by this fungus under SmF. The 83.0 and 95.3 kDa protein bands were present in both C. cubensis cultures. The C. cubensis SSF-SmF-cocktail presented better efficiency in glucose release after 48 h of the alkali-pretreated sugarcane bagasse hydrolysis when compared to those produced by this fungus under either SSF or SmF. This extract showed the highest xylananase/FPase rate and the second highest CMCase/FPase and b-glucosidase/FPase rates among the evaluated extracts, suggesting that these enzymes are the main determinants of this cocktail the efficiency on the alkali pretreated sugarcane bagasse sacchariffication process. These results demonstrated that the enzymes produced by C. cubensis cultivated under SSF and SmF are complementary for the alkali-pretreated sugarcane bagasse enzymatic hydrolysis, since the SSF-SmF-cocktail was more efficient than other extracts produced by this fungus and that the commercial Accellerase®. Therefore, the SSF-SmF-cocktail is a promising alternative for industrial applications.

Graphical abstract

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

Similar content being viewed by others

Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

References

  • Acuña-Argüelles ME, Gutiérrez-Rojas M, Viniegra-González G, Favela-Torres E (1995) Production and properties of three pectinolytic activities produced by Aspergillus niger in submerged and solid-state fermentation. Appl Microbiol Biotechnol 43:808–814

    Article  PubMed  Google Scholar 

  • Andrade LGA, Maitan-Alfenas GP, Morgan T, Gomes KS, Falkoski DL, Alfenas RF, Guimarães VM (2017) Sugarcane bagasse saccharification by purified β-glucosidases from Chrysoporthe cubensis. Biocatal Agric Biotechnol 12(199):205

    Google Scholar 

  • Barrios-González J (2012) Solid-state fermentation: Physiology of solid medium, its molecular basis and applications. Process Biochem 47:175–185

    Article  Google Scholar 

  • Bradford M (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  PubMed  Google Scholar 

  • Champreda V, Mhuantong W, Lekakarn H, Bunterngsook B, Kanokratana P, Zhao X-Q, Zhang F, Inoue H, Fujii T, Eurwilaichitr L (2019) Designing cellulolytic enzyme systems for biorefinery: from nature to application. J Biosci Bioeng 128:637–654

    Article  PubMed  Google Scholar 

  • de Albuquerque MFG, Guimarães VM, de Rezende ST (2021) Use of sugar beet flour and wheat bran as carbon source improves the efficiency of Chrysoporthe cubensis enzymes in sugarcane bagasse saccharification. Bioenerg Res 14:1147–1160

    Article  Google Scholar 

  • de Almeida MN, Falkoski DL, Guimarães VM, Ramos HJO, Visser EM, Maitan-Alfenas GP, De Rezende ST (2013) Characteristics of free endoglucanase and glicosydases multienzyme complex from Fusarium verticillioides. Bioresour Technol 143:413–422

    Article  PubMed  Google Scholar 

  • Dutra TR, Guimarães VM, Varela EM, Fialho LS, Milagres AMF, Falkoski DL, Zanuncio JC, De Rezende ST (2017) A Chrysoporthe cubensis enzyme cocktail produced from a low-cost carbon source with high biomass hydrolysis efficiency. Sci Rep 7:3893

    Article  PubMed  PubMed Central  Google Scholar 

  • Dyballa N, Metzger S (2009) Fast and sensitive colloidal coomassie G-250 staining for proteins in polyacrylamide gels. J vis Exp 30:1431

    Google Scholar 

  • Falkoski DL, Guimarães VM, De Almeida MN, Alfenas AC, Colodette JL, De Rezende ST (2013) Chrysoporthe cubensis: a new source of cellulases and hemicellulases to application in biomass saccharification processes. Bioresour Technol 130:296–305

    Article  CAS  PubMed  Google Scholar 

  • Fanaei MA, Vaziri BM (2009) Modeling of temperature gradients in packed-bed solid-state bioreactors. Chem Eng Process 48:446–451

    Article  CAS  Google Scholar 

  • Ghose TK (1987) Measurement of cellulose activities. Pure Appl Chem 59:257–268

    Article  CAS  Google Scholar 

  • Hernández-Pérez AF, Arruda PV, Felipe MGA (2016) Sugarcane straw as a feedstock for xylitol production by Candida guilliermondii FTI 20037. Braz J Microbiol 47:489–496

    Article  PubMed  PubMed Central  Google Scholar 

  • Holker U, Hofer M, Lenz J (2004) Biotechnological advantages of laboratory-scale solid-state fermentation with fungi. Appl Microbiol Biotechnol 64:175–186

    Article  CAS  PubMed  Google Scholar 

  • 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 4:36

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Inoue H, Kitao C, Yano S, Sawayama S (2016) Production of β-xylosidase from Trichoderma asperellum KIF125 and its application in efficient hydrolysis of pretreated rice straw with fungal cellulase. World J Microbiol Biotechnol 32:186

    Article  PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  • Maitan-Alfenas GP, Visser EM, Alfenas RF, Nogueira BRG, Campos GG, Milagres AF, De Vries RP, Guimarães VM (2015) The influence of pretreatment methods on saccharification of sugarcane bagasse by an enzyme extract from Chrysoporthe cubensis and commercial cocktails: a comparative study. Bioresour Technol 192:670–676

    Article  CAS  PubMed  Google Scholar 

  • Mendonça EHM, Avanci NC, Romano LH, Branco DL, Pádua AX, Ward RJ, Neto AB, Lourenzoni MR (2020) Recombinant xylanase production by Escherichia coli using a non-induced expression system with different nutrient sources. Braz J Chem Eng. https://doi.org/10.1007/s43153-019-00004-x

    Article  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicycilic acid reagent for determination of reducing sugars. Anal Chem 31:426–430

    Article  CAS  Google Scholar 

  • Olsen PO, Alasepp K, Kari J, Cruys-Bagguer N, Borch K, Westh P (2016) Mechanism of product inhibition for Cellobiohydrolase Cel7A during hydrolysis of insoluble cellulose. Biotechnol Bioeng 113:6

    Article  Google Scholar 

  • Rao RS, Jyothi CP, PrakashamRS SPN, Rao LV (2006) Xylitol production from corn fiber and sugarcane bagasse hydrolysates by Candida tropicalis. Bioresour Technol 97:1974–1978

    Article  CAS  PubMed  Google Scholar 

  • Saritha M, Singh S, Tiwari R, Goel R, Nain L (2016) Do cultural conditions induce differential protein expression: Profiling of extracellular proteome of Aspergillus terreus CM20. Microbiol Res 192:73–83

    Article  CAS  Google Scholar 

  • Singhvi MS, Adsul MG, Gokhale DV (2011) Comparative production of cellulases by mutants of Penicillium janthinellum NCIM 1171 and its application in hydrolysis of Avicel and cellulose. Bioresour Technol 102:6569–6572

    Article  CAS  PubMed  Google Scholar 

  • Subramaniyam R, Vimala R (2004) Solid state and submerged fermentation for the production of bioactive substances: a comparative study. Int J Sci Nat 3:480–486

    Google Scholar 

  • Tavares MP, Morgan T, Gomes RF, Rodrigues MQRB, Castro-Borges W, de Rezende ST, Mendes TAO, Guimarães VM (2021) Secretomic insight into the biomass hydrolysis potential of the phytopathogenic fungus Chrysoporthe cubensis. J Proteom 236:104121

    Article  CAS  Google Scholar 

  • Venegas IM, Fuentes-Hernández J, Garcia-Rivero M, Martinez-Treujillo A (2013) Characteristics of Aspergillus niger xylanases produced on ricehusk and wheat bran in submerged culture and solid-statefermentation for an applicability proposal. Int J Food Sci Technol 48:1798–1807

    Article  Google Scholar 

  • Vicari KJ, Tallam SS, Shatova T, Joo KK, Scarlata CJ, Humbird D, Wolfrum EJ, Beckham GT (2012) Uncertainty in techno-economic estimates of cellulosic ethanol production due to experimental measurement uncertainty. Biotechnol Biofuels 5(1):1-12

  • Viniegra-gonzález G, Favela-Torres E, Aguilar CN, Rómero-Gomez SJ, Díaz-Godínez G, Augur C (2003) Advantages of fungal enzyme production in solid state over liquid fermentation systems. Biochem Eng J 13:157–167

    Article  Google Scholar 

  • Visser EM, Falkoski DL, De Almeida MN, Maitan-Alfenas GP, Guimarães VM (2013) Production and application of an enzyme blend from Chrysoporthe cubensis and Penicillium pinophilum with potential for hydrolysis of sugarcane bagasse. Bioresour Technol 144:587–594

    Article  CAS  PubMed  Google Scholar 

  • Visser EM, Leal TF, De Almeida MN, Guimarães VM (2015) Increased enzymatic hydrolysis of sugarcane bagasse from enzyme recycling. Biotechnol Biofuels 8:5

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang X, Tu M, Paice MG (2011) Routes to potential bioproducts from lignocellulosic biomass lignin and hemicelluloses. Bioenergy Res 4:246–257

    Article  Google Scholar 

Download references

Acknowledgements

We thank Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), Conselho Nacional de Pesquisa e Desenvolvimento Tecnológico (CNPq) for the financial support and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for providing scholarships. We would like to thank Thamires Felisberto Pereira Dutra for cooperating in the experiments execution.

Author information

Authors and Affiliations

  1. Departamento de Bioquímica e Biologia Molecular (BIOAGRO), Universidade Federal de Viçosa, 36570-000, Viçosa, Minas Gerais, Brazil

    Thiago Rodrigues Dutra, Valéria Monteze Guimarães & Sebastião Tavares de Rezende

  2. Universidade Nova de Lisboa/CEDOC - Chronic Diseases Research Center, 1150-190, Lisbon, Portugal

    Ednilson Mascarenhas Varela

  3. Departamento de Entomologia/BIOAGRO, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brazil

    Mariana Furtado Granato de Albuquerque

  4. Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, 35400-000, Ouro Preto, Minas Gerais, Brazil

    Juan Diego Rios Díez

  5. Departamento de Fitopatologia, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brazil

    Acelino Couto Alfenas

Authors
  1. Thiago Rodrigues Dutra
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Valéria Monteze Guimarães
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ednilson Mascarenhas Varela
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mariana Furtado Granato de Albuquerque
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Juan Diego Rios Díez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Acelino Couto Alfenas
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sebastião Tavares de Rezende
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Thiago Rodrigues Dutra.

Ethics declarations

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dutra, T.R., Guimarães, V.M., Varela, E.M. et al. A new approach for Chrysoporthe cubensis cellulolytic cocktail production using solid and submerged-state fermentation. Braz. J. Chem. Eng. 40, 359–366 (2023). https://doi.org/10.1007/s43153-023-00309-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43153-023-00309-y

Keywords

Search

Navigation