Abstract
Objectives
The use of endo-arabinanase from Bacillus licheniformis (ABNase) for sugarcane saccharification has been evaluated by enzyme immobilization and commercial cocktail supplement with the immobilized heterologous protein.
Results
Biochemical characterization of the purified ABNase showed that the catalytic activity was strongly inhibited by 5 mM Cu2+, Zn2+ or Fe3+. The optimum pH and temperature for activity were 5.5–6.5 and 35–40 °C, respectively. The enzyme stability increased 128-fold when immobilized with glyoxyl agarose, and the hydrolysis of pretreated sugar cane biomass increased by 15 % when a commercial enzyme cocktail was supplemented with immobilized ABNase.
Conclusion
Pectin hydrolysis by recombinant ABNase plays a role in the effective application of enzymatic cocktails for biomass saccharification.
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Acknowledgements
This work was supported by research Grants 2013/14454-3, 2010/18850-2 (FAPESP) and 153501/2012-0 (CNPq).
Supporting information
Supplementary Fig. 1—Purification of soluble recombinant ABNase protein by affinity chomatography. 12 % SDS Page of ABNase purification showing the (MM) molecular weight markers (Fermentas, molecular weight indicated); (0h) no IPTG induction; (4h) of IPTG induction; (S) soluble fraction and (F) fractions eluted from the column.
Supplementary Fig. 2—Temperature and pH profile of arabinanase. (A) Surface response and (B) Contour curves generated in the central composite rotatable design (CCRD) to determine the optimum pH and temperature. The enzymatic reactions were carried out at different pHs (4.0, 4.9, 7.0, 9.1 and 10.0) in 0.1 M citric acid / 0.1 M Na2HPO4 / 0.1 M glycine buffer and a range of temperatures (20, 28.7, 50, 71.3 e 80°C).
Supplementary Table 1—Matrix of the CCRD (Central Composite Rotational Design) to determine the optimal temperature and pH of arabinanase from Bacillus licheniformis.
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Machado, C.B., Citadini, A.P., Goldbeck, R. et al. Increased biomass saccharification by supplementation of a commercial enzyme cocktail with endo-arabinanase from Bacillus licheniformis . Biotechnol Lett 37, 1455–1462 (2015). https://doi.org/10.1007/s10529-015-1818-0
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DOI: https://doi.org/10.1007/s10529-015-1818-0