The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse

Lorena Cardoso Cintra, Isabelle Cristine da Costa, Izadora Cristina Moreira de Oliveira, Amanda Gregorim Fernandes, Syd Pereira Faria, Rosália Santos Amorin Jesuíno, Maria Cristina Ravanal, Jaime Eyzaguirre, Luiz Pereira Ramos, Fabrícia Paula de Faria, Cirano José Ulhoa

Resultado de la investigación: Article

Resumen

To increase the efficiency of enzyme cocktails in deconstructing cellulose and hemicelluloses present in the plant cell wall, a combination of enzymes with complementary activities is required. Xylan is the main hemicellulose component of energy crops and for its complete hydrolysis a system consisting of several enzymes acting cooperatively, including endoxylanases (XYN), β-xylosidases (XYL) and α-L-arabinofuranosidases (ABF) is necessary. The current work aimed at evaluating the effect of recombinant hemicellulolytic enzymes on the enzymatic hydrolysis of steam-exploded sugarcane bagasse (SEB). One recombinant endoxylanase (HXYN2) and one recombinant β-xylosidase (HXYLA) from Humicola grisea var thermoidea, together with an α-L-arabinofuranosidase (AFB3) from Penicillium pupurogenum, all produced in Pichia pastoris, were used to formulate an efficient enzyme mixture for SEB hydrolysis using a 23 Central Composite Rotatable Design (CCRD). The most potent enzyme for SEB hydrolysis was ABF3. Subsequently, the optimal enzyme mixture was used in combination with commercial cellulases (Accellerase 1500), either simultaneously or in sequential experiments. The supplementation of Accellerase 1500 with hemicellulases enhanced the glucose yield from SEB hydrolysis by 14.6%, but this effect could be raised to 50% when hemicellulases were added prior to hydrolysis with commercial cellulases. These results were supported by scanning electron microscopy, which revealed the effect of enzymatic hydrolysis on SEB fibers. Our results show the potential of complementary enzyme activities to improve enzymatic hydrolysis of SEB, thus improving the efficiency of the hydrolytic process.

Idioma originalEnglish
Número de artículo109447
PublicaciónEnzyme and Microbial Technology
Volumen133
DOI
EstadoPublished - feb 2020

Huella dactilar

Saccharum
Bagasse
Enzymatic hydrolysis
Steam
Hydrolysis
Enzymes
Xylosidases
Endo-1,4-beta Xylanases
Cellulases
Xylans
Enzyme activity
Pichia
hemicellulase
bagasse
Penicillium
Cellulose
Plant Cells
Crops
Glucose
Electron Scanning Microscopy

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

Citar esto

Cintra, L. C., da Costa, I. C., de Oliveira, I. C. M., Fernandes, A. G., Faria, S. P., Jesuíno, R. S. A., ... Ulhoa, C. J. (2020). The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse. Enzyme and Microbial Technology, 133, [109447]. https://doi.org/10.1016/j.enzmictec.2019.109447
Cintra, Lorena Cardoso ; da Costa, Isabelle Cristine ; de Oliveira, Izadora Cristina Moreira ; Fernandes, Amanda Gregorim ; Faria, Syd Pereira ; Jesuíno, Rosália Santos Amorin ; Ravanal, Maria Cristina ; Eyzaguirre, Jaime ; Ramos, Luiz Pereira ; de Faria, Fabrícia Paula ; Ulhoa, Cirano José. / The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse. En: Enzyme and Microbial Technology. 2020 ; Vol. 133.
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abstract = "To increase the efficiency of enzyme cocktails in deconstructing cellulose and hemicelluloses present in the plant cell wall, a combination of enzymes with complementary activities is required. Xylan is the main hemicellulose component of energy crops and for its complete hydrolysis a system consisting of several enzymes acting cooperatively, including endoxylanases (XYN), β-xylosidases (XYL) and α-L-arabinofuranosidases (ABF) is necessary. The current work aimed at evaluating the effect of recombinant hemicellulolytic enzymes on the enzymatic hydrolysis of steam-exploded sugarcane bagasse (SEB). One recombinant endoxylanase (HXYN2) and one recombinant β-xylosidase (HXYLA) from Humicola grisea var thermoidea, together with an α-L-arabinofuranosidase (AFB3) from Penicillium pupurogenum, all produced in Pichia pastoris, were used to formulate an efficient enzyme mixture for SEB hydrolysis using a 23 Central Composite Rotatable Design (CCRD). The most potent enzyme for SEB hydrolysis was ABF3. Subsequently, the optimal enzyme mixture was used in combination with commercial cellulases (Accellerase 1500), either simultaneously or in sequential experiments. The supplementation of Accellerase 1500 with hemicellulases enhanced the glucose yield from SEB hydrolysis by 14.6{\%}, but this effect could be raised to 50{\%} when hemicellulases were added prior to hydrolysis with commercial cellulases. These results were supported by scanning electron microscopy, which revealed the effect of enzymatic hydrolysis on SEB fibers. Our results show the potential of complementary enzyme activities to improve enzymatic hydrolysis of SEB, thus improving the efficiency of the hydrolytic process.",
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author = "Cintra, {Lorena Cardoso} and {da Costa}, {Isabelle Cristine} and {de Oliveira}, {Izadora Cristina Moreira} and Fernandes, {Amanda Gregorim} and Faria, {Syd Pereira} and Jesu{\'i}no, {Ros{\'a}lia Santos Amorin} and Ravanal, {Maria Cristina} and Jaime Eyzaguirre and Ramos, {Luiz Pereira} and {de Faria}, {Fabr{\'i}cia Paula} and Ulhoa, {Cirano Jos{\'e}}",
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Cintra, LC, da Costa, IC, de Oliveira, ICM, Fernandes, AG, Faria, SP, Jesuíno, RSA, Ravanal, MC, Eyzaguirre, J, Ramos, LP, de Faria, FP & Ulhoa, CJ 2020, 'The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse', Enzyme and Microbial Technology, vol. 133, 109447. https://doi.org/10.1016/j.enzmictec.2019.109447

The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse. / Cintra, Lorena Cardoso; da Costa, Isabelle Cristine; de Oliveira, Izadora Cristina Moreira; Fernandes, Amanda Gregorim; Faria, Syd Pereira; Jesuíno, Rosália Santos Amorin; Ravanal, Maria Cristina; Eyzaguirre, Jaime; Ramos, Luiz Pereira; de Faria, Fabrícia Paula; Ulhoa, Cirano José.

En: Enzyme and Microbial Technology, Vol. 133, 109447, 02.2020.

Resultado de la investigación: Article

TY - JOUR

T1 - The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse

AU - Cintra, Lorena Cardoso

AU - da Costa, Isabelle Cristine

AU - de Oliveira, Izadora Cristina Moreira

AU - Fernandes, Amanda Gregorim

AU - Faria, Syd Pereira

AU - Jesuíno, Rosália Santos Amorin

AU - Ravanal, Maria Cristina

AU - Eyzaguirre, Jaime

AU - Ramos, Luiz Pereira

AU - de Faria, Fabrícia Paula

AU - Ulhoa, Cirano José

PY - 2020/2

Y1 - 2020/2

N2 - To increase the efficiency of enzyme cocktails in deconstructing cellulose and hemicelluloses present in the plant cell wall, a combination of enzymes with complementary activities is required. Xylan is the main hemicellulose component of energy crops and for its complete hydrolysis a system consisting of several enzymes acting cooperatively, including endoxylanases (XYN), β-xylosidases (XYL) and α-L-arabinofuranosidases (ABF) is necessary. The current work aimed at evaluating the effect of recombinant hemicellulolytic enzymes on the enzymatic hydrolysis of steam-exploded sugarcane bagasse (SEB). One recombinant endoxylanase (HXYN2) and one recombinant β-xylosidase (HXYLA) from Humicola grisea var thermoidea, together with an α-L-arabinofuranosidase (AFB3) from Penicillium pupurogenum, all produced in Pichia pastoris, were used to formulate an efficient enzyme mixture for SEB hydrolysis using a 23 Central Composite Rotatable Design (CCRD). The most potent enzyme for SEB hydrolysis was ABF3. Subsequently, the optimal enzyme mixture was used in combination with commercial cellulases (Accellerase 1500), either simultaneously or in sequential experiments. The supplementation of Accellerase 1500 with hemicellulases enhanced the glucose yield from SEB hydrolysis by 14.6%, but this effect could be raised to 50% when hemicellulases were added prior to hydrolysis with commercial cellulases. These results were supported by scanning electron microscopy, which revealed the effect of enzymatic hydrolysis on SEB fibers. Our results show the potential of complementary enzyme activities to improve enzymatic hydrolysis of SEB, thus improving the efficiency of the hydrolytic process.

AB - To increase the efficiency of enzyme cocktails in deconstructing cellulose and hemicelluloses present in the plant cell wall, a combination of enzymes with complementary activities is required. Xylan is the main hemicellulose component of energy crops and for its complete hydrolysis a system consisting of several enzymes acting cooperatively, including endoxylanases (XYN), β-xylosidases (XYL) and α-L-arabinofuranosidases (ABF) is necessary. The current work aimed at evaluating the effect of recombinant hemicellulolytic enzymes on the enzymatic hydrolysis of steam-exploded sugarcane bagasse (SEB). One recombinant endoxylanase (HXYN2) and one recombinant β-xylosidase (HXYLA) from Humicola grisea var thermoidea, together with an α-L-arabinofuranosidase (AFB3) from Penicillium pupurogenum, all produced in Pichia pastoris, were used to formulate an efficient enzyme mixture for SEB hydrolysis using a 23 Central Composite Rotatable Design (CCRD). The most potent enzyme for SEB hydrolysis was ABF3. Subsequently, the optimal enzyme mixture was used in combination with commercial cellulases (Accellerase 1500), either simultaneously or in sequential experiments. The supplementation of Accellerase 1500 with hemicellulases enhanced the glucose yield from SEB hydrolysis by 14.6%, but this effect could be raised to 50% when hemicellulases were added prior to hydrolysis with commercial cellulases. These results were supported by scanning electron microscopy, which revealed the effect of enzymatic hydrolysis on SEB fibers. Our results show the potential of complementary enzyme activities to improve enzymatic hydrolysis of SEB, thus improving the efficiency of the hydrolytic process.

KW - Enzymatic hydrolysis

KW - Recombinant hemicellulases

KW - Sugarcane bagasse

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U2 - 10.1016/j.enzmictec.2019.109447

DO - 10.1016/j.enzmictec.2019.109447

M3 - Article

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