Solid-liquid extraction of cellulases from fungal solid-state cultivation in a packed bed bioreactor

Samuel Pratavieira de Oliveira; Natalia Alvarez Rodrigues; Priscila Aparecida Casciatori-Frassatto; Fernanda Perpétua Casciatori

Korean Journal of Chemical Engineering, Vol.37, No.9, 1530-1540, September, 2020

DOI10.1007/s11814-020-0579-1 Export Citation

Solid-liquid extraction of cellulases from fungal solid-state cultivation in a packed bed bioreactor

Samuel Pratavieira de Oliveira, Natalia Alvarez Rodrigues, Priscila Aparecida Casciatori-Frassatto¹, and Fernanda Perpétua Casciatori^†

Department of Chemical Engineering, Federal University of São Carlos (UFSCar), Rodovia Washington Luiz km 235 (SP 310), 13565-905, São Carlos - SP, Brazil
¹Department of Chemistry and Environmental Sciences, Institute of Biosciences, Letters and Exact Sciences, São Paulo State University (UNESP), Cristóvão Colombo 2265, 15054-000, São José do Rio Preto - SP, Brazil

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Cellulases are enzymes required for the production of second-generation ethanol (E2G) via biochemical route. The current paper reports the development of an apparatus for solid-liquid extraction of cellulases from solidstate fermentation (SSF) carried out in a packed bed bioreactor (PBB), operated as batch and as semicontinuous. The case study was the cultivation of Myceliophthora thermophila I-1D3b on sugarcane bagasse (SCB) and wheat bran (WB) (7 : 3 w/w). The current work integrates the PBB to the first downstream step for recovering the enzymes produced by SSF. The substrate was inoculated and packed into the modules that composed the PBB. The fermentation occurred at 45 oC and air was supplied with flow rate of 350 L/h. At the end of the cultivations, each module was placed in an extraction column, a dynamic closed system in which distilled water was circulated and made to percolate the cultivated material. Variables tested were volume of water per mass of substrate, water flow rate and time of percolation. Higher contact time (120min) and higher flow rate (2.4m³/h) allowed us to recover up to 85% of total enzyme activity by percolation. Lower volume (20mL/g) provides higher titer extract. The apparatus showed to be promising for SSF downstream, mainly for semicontinuous operation of PPBs.

Keywords:Fixed Bed;Downstream;Cellulolytic Enzymes;Bioethanol;Biorefineries

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[1] Travaini R, Otero MDM, Coca M, Da-Silva R, Bolado S, Bioresour. Technol., 133, 332 (2013)

[2] Florencio C, Badino AC, Farinas CS, Quim. Nova, 40, 1082 (2017)

[3] Pandey A, Soccol CR, Nigam P, Soccol VT, Bioresour. Technol., 74(1), 69 (2000)

[4] Mishima D, Tateda M, Ike M, Fujita M, Bioresour. Technol., 97(16), 2166 (2006)

[5] Farinas CS, Florencio C, Badino AC, in Cellulases. Methods in molecular biology, Humana Press, New York (2018).

[6] Chandra MS, Viswanath B, Reddy BR, Indian J. Microbiol., 47, 323 (2007)

[7] Mitchell DA, Krieger N, Berovic M, Solid-state fermentation bioreactors: fundamentals, design and operation, Springer-Verlag, Berlin (2006).

[8] da Silva R, Lago ES, Merheb CW, Macchione MM, Park YK, Braz. J. Microbiol., 36, 235 (2005)

[9] Casciatori FP, Bueck A, Thomeo JC, Tsotsas E, Chem. Eng. J., 287, 103 (2016)

[10] Casciatori FP, Thomeo JC, in Topics in waste treatment and environment, Novas Edicoes Academicas, Saarbrucken (2015).

[11] Mitchell DA, Cunha LEN, Machado AVL, Luz LFL, Krieger N, Biochem. Eng. J., 48, 195 (2010)

[12] Pirota RDPB, Miotto LS, Delabona PS, Farinas CS, Braz. J. Chem. Eng., 30, 177 (2013)

[13] Zanelato AI, Shiota VM, Gomes E, Thomeo JC, Braz. J. Microbiol., 43, 1536 (2012)

[14] Perez CL, Casciatori FP, Thomeo JC, Chem. Eng. J., 361, 1142 (2019)

[15] Bradford MM, Anal. Biochem., 72, 248 (1976)

[16] Ghose TK, Pure Appl. Chem., 59, 257 (1987)

[17] Miller GL, Anal. Chem., 31, 426 (1959)

[18] Calixto LO, UFSCar (2019).

[19] Casciatori FP, Laurentino CL, Lopes KCM, Souza AG, Thomeo JC, Int. J. Food Prop., 16, 1578 (2013)

[20] Casciatori FP, Laurentino CL, Taboga SR, Casciatori PA, Thomeo JC, Chem. Eng. J., 255, 214 (2014)

[21] Ichiba IT, UFSCar (2019).

[22] Fenila F, Shastri Y, Resource-Efficient Technol., 2, S96 (2016)