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β-Glucosidase production by Trichoderma reesei and Thermoascus aurantiacus by solid state cultivation and application of enzymatic cocktail for saccharification of sugarcane bagasse

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Abstract

For degradation of sugarcane bagasse (SCB), several enzymes are needed but β-glucosidase is rate limiting in cellulose hydrolysis. Since different microorganisms synthetize characteristic pool of enzymes, mixing extracts produced by different species may increase hydrolytic efficiency due to synergism between enzymes in cocktails. This paper reports the study of β-glucosidase production in solid state cultivation (SSC) of two filamentous fungi, thermophilic Thermoascus aurantiacus and mesophilic Trichoderma reesei, and application of the enzymatic extracts on non-pretreated SCB saccharification. Enzyme extract obtained from the thermophilic fungus presented higher β-glucosidase and FPU activities (1.8 U/mL and 10 FPU/mL) than the one from mesophilic (0.2 U/mL and 6 FPU/mL). Optimal SCB hydrolysis was achieved when applying enzymatic cocktail composed of equal volumes of both fungal extracts (3.6 FPU/gSCB, filter paper units per gram SCB, 2.25 FPU/gSCB provided by extract from T. aurantiacus and 1.35 FPU/gSCB from T. reesei) at 65 °C. The hydrolysis yield applying the enzyme cocktail, 124 mg total reducing sugars (TRS) per gSCB, was higher than any yield achieved when using the enzyme extracts separately (105 mgTRS/gSCB using 12.5 FPU per gSCB from T. aurantiacus at 65 °C; 79 mgTRS/gSCB using 7.5 FPU per gSCB from T. reesei at 45 °C). Therefore, the use of the cocktail (3.6 FPU/gSCB) at 65 °C released 18 and 57% more TRS respectively than when extracts from T. aurantiacus or from T. reesei were applied alone, respectively, even reducing enzyme load (FPU) by 70%, corroborating the synergistic effect when both extracts are used together.

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Acknowledgments

The authors would like to acknowledge the National Council for the Improvement of Higher Education (CAPES) and the São Paulo Research Foundation (FAPESP) for their financial support and scholarships [grant numbers 2010/12624-0, 2011/21239-6, 2011/07453-5, 2012/02768-0, 2013/01756-1, 2017/16482-5, 2018/00996-2] and the National Council for Scientific and Technological Development (CNPQ) [grant number 426578/2016-3].

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Authors and Affiliations

  1. Department of Chemistry and Environmental Sciences, Institute of Biosciences, Letters and Exact Sciences, São Paulo State University (UNESP), Cristóvão Colombo 2265, São José do Rio Preto, SP, 15054-000, Brazil

    Priscila Aparecida Casciatori Frassatto, Maurício Boscolo & Roberto da Silva

  2. Department of Chemical Engineering, Federal University of São Carlos (UFSCar), Rodovia Washington Luís km 235 (SP 310), São Carlos, SP, 13565-905, Brazil

    Fernanda Perpétua Casciatori

  3. Department of Food Engineering and Technology, Institute of Biosciences, Letters and Exact Sciences, São Paulo State University (UNESP), Cristóvão Colombo 2265, São José do Rio Preto, SP, 15054-000, Brazil

    João Cláudio Thoméo

  4. Department of Biology, Institute of Biosciences, Letters and Exact Sciences, São Paulo State University (UNESP), Cristóvão Colombo 2265, São José do Rio Preto, SP, 15054-000, Brazil

    Eleni Gomes

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  1. Priscila Aparecida Casciatori Frassatto
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Highlights

• Thermophilic fungus yielded more β-glucosidase than the mesophilic one, in a shorter time;

• Fungal biomass was well estimated based on total protein content by the Kjeldahl method;

• β-Glucosidase activity production was strongly and directly associated with fungal growth;

• Enzymes in cocktail have a synergistic effect on the hydrolysis of sugarcane bagasse.

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Frassatto, P.A.C., Casciatori, F.P., Thoméo, J.C. et al. β-Glucosidase production by Trichoderma reesei and Thermoascus aurantiacus by solid state cultivation and application of enzymatic cocktail for saccharification of sugarcane bagasse. Biomass Conv. Bioref. 11, 503–513 (2021). https://doi.org/10.1007/s13399-020-00608-1

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