Most of the corn-ethanol plants use dry-mill process and generate large volume of distillers dried grains with solubles (DDGS) as the co-product. The DDGS is a promising feedstock with rich content of cellulose and xylan. The goal of this study was to optimize cellulase and xylanase production by Trichoderma reesei (NRRL 6156) using DDGS as the carbon source at shake-flask and validate the results at benchtop bioreactors. The fermentation medium containing varied amounts of dilute-acid pretreated DDGS, yeast extract, and peptone were optimized for maximal enzyme production at 30 °C, pH 5.0 and 180 rpm with 3% (v/v) inoculum using Box-Behnken response surface methodology (RSM). Co-production of xylanase and cellulase enzymes was not achieved in the same medium; higher DDGS hydrolysate favored cellulase enzyme only while lower amount induced the xylanase enzyme. Based on the RSM results, maximal xylanase of 18.5 IU/mL was predicted with 5% DDGS hydrolysate, 0.1 g/L yeast extract, and 2 g/L peptone, while maximal cellulase of 1.1 IU/mL was predicted with 20% DDGS hydrolysate, 1 g/L yeast extract, and 0.5 g/L peptone. Of the two developed models from shake-flasks, xylanase model validated successfully the fermenter results giving 18.7 IU/mL of experimental xylanase in 1.5-L working volume bioreactor.
This is a preview of subscription content, log in to check access.
Buy single article
Instant unlimited access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Camassola, M., Dillon, A.J.P.: Production of cellulases and hemicellulases by Pencillium echinulatum grown on pretreated sugar cane bagasse and wheat bran in solid-state fermentation. J. Appl. Microbiol. 10, 2196–2204 (2007)
Ortiz, G.E., Guitart, M.E., Cavalitto, S.F., Alberto, E.O., Fernandez-Lahore, M., Blasco, M.: Characterization, optimization, and scale-up of cellulases production by Trichoderma reesei cbs 836.91 in solid-state fermentation using agro-industrial products. Bioproc. Biosys. Eng. 38, 2117–2128 (2015)
Li, S., Tang, B., Xu, Z., Chen, T., Liu, L.: Fermentation optimization and unstructured kinetic model for cellulase production by Rhizopus stolonifer var. reflexus TP-02 on agriculture by-products. Appl. Biochem. Biotechnol. 177, 1589–1606 (2015)
Cao, Y., Meng, D., Lu, J., Long, J.: Statistical optimization of xylanase production by Aspergillus niger AN-13 under submerged fermentation using response surface methodology. Afr. J. Biotechnol. 7(5), 631–638 (2008)
Sukumaran, R.K., Singhania, R.R., Mathew, G.M., Pandey, A.: Cellulase production using biomass feed stock and its application in lignocellulose saccharification for bio-ethanol production. Renew. Energy 34, 421–424 (2009)
Ghoshal, G., Banerjee, U.C., Shivhare, U.S.: Xylanase production by Penicillium citrinum in laboratory scale stirred tank reactor. Chem. Biochem. Eng. J. 28(3), 399–408 (2014)
Schaffner, D.W., Toledo, R.T.: Cellulase production by Trichoderma reesei when cultured on xylose-based media supplemented with sorbose. Biotechnol. Bioeng. 37, 12–16 (1991)
Salmon, D.N.X., Spier, M.R., Soccol, C.R., Vandenberghe, L.P. de Souza, Montibeller, V.W., Bier, M.C.J., Faraco, V.: Analysis of inducers of xylanase and cellulase activities production by Ganoderma applanatum LPB MR-56. Fungal Biol. 118, 655–662 (2014)
Ximenes, E.A., Dien, B.S., Ladisch, M.R., Mosier, N., Cotta, M.A., Li, X.L.: Enzyme production by industrially relevant fungi cultured on coproduct from corn dry grind ethanol plants. Appl. Biochem. Biotechnol. 136–140, 171–184 (2007)
Wang, X., Wang, Y., Wang, B., Blaschek, H., Feng, H., Li, Z.: Biobutanol production from fiber-enhanced DDGS pretreated with electrolyzed water. Renew. Energy 52, 16–22 (2013)
Kim, Y., Hendrickson, R., Mosier, N.S., Ladisch, M.R., Bals, B., Balan, V., Dale, B.E.: Enzyme hydrolysis and ethanol fermentation of liquid hot water and AFEX pretreated distillers’ grains at high-solids loadings. Bioresour. Technol. 99, 5206–5215 (2008)
Lio, J.Y., Wang, T.: Solid-state fermentation of soybean and corn processing coproducts for potential feed development. J. Agric. Food Chem. 60, 7702–7709 (2012)
Yang, S., Lio, J.Y., Wang, T.: Evaluation of enzyme activity and fiber content of soybean cotyledon fiber and distiller’s dried grains with solubles by solid state fermentation. Appl. Biochem. Biotechnol. 167, 109–121 (2012)
Muthuvelayudham, R., Viruthagiri, T.: Fermentative production and kinetics of cellulase protein on Trichoderma reesei using sugarcane bagasse and rice straw. Afr. J. Biotechnol. 5(20), 1873–1881 (2006)
Laxmi, G.S., Sathish, T., Rao, ChS, Brahmaiah, P., Hymavathi, M., Prakasham, R.S.: Palm fiber as novel substrate for enhanced xylanase production by isolated Aspergillus sp. RSP-6. Curr. Trends Biotechnol. Pharm. 2(3), 447–455 (2008)
Li, X.L., Dien, B.S., Cotta, M.A., Wu, Y.V., Saha, B.C.: Profile of enzyme production by Trichoderma reesei grown on corn fiber fractions. Appl. Biochem. Biotechnol. 121–124, 321–334 (2005)
Li, J., Zhou, P., Liu, H., Lin, J., Gong, Y., Xiao, W., Liu, Z.: Monosaccharides and ethanol production from superfine ground sugarcane bagasse using enzyme coctail. BioResources 9(2), 2529–2540 (2014)
Bayer, E.A., Shoham, Y., Lamed, R.: Cellulose-decomposing bacteria and their enzyme systems. In: Dworkin, M., Falkow, S., Rosenberg, E., Schleifer, K.H., Stackebrandt, E. (eds.) The prokaryotes: a handbook on the biology of bacteria, pp. 578–617. Springer, New York (2006)
Vitcosque, G.L., Fonseca, R.F., Rodriguez-Zuniga, U.F., Neto, V.B., Couri, S., Farinas, C.S.: Production of biomass-degrading multienzyme complexes under solid-state fermentation of soybean meal using a bioreactor. Enzyme Res. (2012). https://doi.org/10.1155/2012/248983
Wang, H.C., Chen, Y.C., Hseu, R.S.: Purification and characterization of a cellulolytic multienzyme complex produced by Neocallimastix patriciarum J11. Biochem. Biophys. Res. Comm. 451(2), 190–195 (2014)
Cekmecelioglu, D., Demirci, A.: A statistical optimization study on dilute sulfuric acid pretreatment of distillers dried grains with solubles (DDGS) as a potential feedstock for fermentation applications. Waste Biomass Valoriz. (2018). https://doi.org/10.1007/s12649-018-0376-9
Box, G.E.P., Behnken, D.W.: Some new three level designs for the study of quantitative variables. Technometrics 2, 455–475 (1960)
Miller, G.L.: Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal. Chem. 31, 426–428 (1959)
Mandels, M., Medeiros, J.E., Andreotti, R.E., Bissett, F.H.: Enzymatic hydrolysis of cellulose: Evaluation of cellulase culture filtrates under use conditions. Biotechnol. Bioeng. (1981). https://doi.org/10.1002/bit.260230907
Ghose, T.K.: Measurement of cellulase activities. Pure Appl. Chem. 59(2), 257–268 (1987)
Adney, B., Baker, J.: Measurement of cellulase activities. Laboratory Analytical Procedure (LAP), National Renewable Energy Laboratory. Technical Report NREL/TP-510-42628 (1996)
Eveleigh, D.E., Mandels, M., Andreotti, R., Roche, C.: Measurement of saccharifying cellulase. Biotechnol. Biofuels. (2009). https://doi.org/10.1186/1754-6834-2-21
Domingues, F.C., Queiroz, J.A., Cabral, J.M.S., Fonseca, L.P.: The influence of culture conditions on mycelial structure and cellulase production by Trichoderma reesei Rut C-30. Enzyme Microbiol. Technol. 26, 394–401 (2000)
This study was supported in-part by USDA Northeast Sun Grant Initiative (NE-SGI) Competitive Grants Program, the USDA National Institute of Food and Agriculture Federal Appropriations under Project PEN04671 and Accession number 1017582 and the Department of Science Fellowships and Grant Programmes of Scientific and Technological Research Council of Turkey (TUBITAK-BIDEB) (Grant No.1059B191500892) by providing scholarship to Deniz Cekmecelioglu.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Cekmecelioglu, D., Demirci, A. Production of Cellulase and Xylanase Enzymes Using Distillers Dried Grains with Solubles (DDGS) by Trichoderma reesei at Shake-Flask Scale and the Validation in the Benchtop Scale Bioreactor. Waste Biomass Valor (2020). https://doi.org/10.1007/s12649-020-00934-5
- Submerged fermentation
- Trichoderma reesei