| Full text | |
| Author(s): Show less - |
dos Santos, Camila Ramos
[1]
;
de Giuseppe, Priscila Oliveira
[1]
;
Moreira de Souza, Flavio Henrique
[1]
;
Zanphorlin, Leticia Maria
[1]
;
Domingues, Mariane Noronha
[1]
;
Siqueira Pirolla, Renan Augusto
[1]
;
Honorato, Rodrigo Vargas
[2]
;
Costa Tonoli, Celisa Caldana
[2]
;
Bueno de Morais, Mariana Abrahao
[1]
;
de Matos Martins, Vanesa Peixoto
[2]
;
Fonseca, Lucas Miranda
[1]
;
Buchli, Fernanda
[1]
;
Lopes de Oliveira, Paulo Sergio
[2]
;
Gozzo, Fabio Cesar
[3]
;
Murakami, Mario Tyago
[1]
Total Authors: 15
|
| Affiliation: | [1] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Bioethanol Sci & Technol Lab CTBE, BR-13083970 Campinas, SP - Brazil
[2] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Biosci Natl Lab LNBio, BR-13083970 Campinas, SP - Brazil
[3] Univ Estadual Campinas, Inst Chem, Dalton Mass Spectrometry Lab, BR-13083861 Campinas, SP - Brazil
Total Affiliations: 3
|
| Document type: | Journal article |
| Source: | BIOTECHNOLOGY FOR BIOFUELS; v. 11, AUG 11 2018. |
| Web of Science Citations: | 2 |
| Abstract | |
Background: Arabinoxylan is an abundant polysaccharide in industrially relevant biomasses such as sugarcane, corn stover and grasses. However, the arabinofuranosyl di-substitutions that decorate the xylan backbone are recalcitrant to most known arabinofuranosidases (Abfs). Results: In this work, we identified a novel GH51 Abf (XacAbf51) that forms trimers in solution and can cope efficiently with both mono- and di-substitutions at terminal or internal xylopyranosyl units of arabinoxylan. Using mass spectrometry, the kinetic parameters of the hydrolysis of 3(3)-alpha-L-arabinofuranosyl-xylotetraose and 2(3),3(3)-di-alpha-L-arabinofuranosyl-xylotetraose by XacAbf51 were determined, demonstrating the capacity of this enzyme to cleave arabinofuranosyl linkages of internal mono- and di-substituted xylopyranosyl units. Complementation studies of fungal enzyme cocktails with XacAbf51 revealed an increase of up to 20% in the release of reducing sugars from pretreated sugarcane bagasse, showing the biotechnological potential of a generalist GH51 in biomass saccharification. To elucidate the structural basis for the recognition of internal di-substitutions, the crystal structure of XacAbf51 was determined unveiling the existence of a pocket strategically arranged near to the - 1 subsite that can accommodate a second arabinofuranosyl decoration, a feature not described for any other GH51 Abf structurally characterized so far. Conclusions: In summary, this study reports the first kinetic characterization of internal di-substitution release by a GH51 Abf, provides the structural basis for this activity and reveals a promising candidate for industrial processes involving plant cell wall depolymerization. (AU) | |
| FAPESP's process: | 13/13309-0 - Studies of the structural and functional behavior of enzymes evolutionarily specialized in the degradation of plant biomass with potential biotechnological applications |
| Grantee: | Mário Tyago Murakami |
| Support type: | Regular Research Grants |
| FAPESP's process: | 14/17264-3 - New frontiers in structural proteomics: characterizing protein and protein complex structures by mass spectrometry |
| Grantee: | Fabio Cesar Gozzo |
| Support type: | Research Projects - Thematic Grants |
| FAPESP's process: | 15/26982-0 - Exploring novel strategies for depolymerization of plant cell-wall polysaccharides: from structure, function and rational design of glycosyl hydrolases to biological implications and potential biotechnological applications |
| Grantee: | Mário Tyago Murakami |
| Support type: | Research Projects - Thematic Grants |
| FAPESP's process: | 14/07135-1 - Multi-user equipament approved in grant 2013/13309-0: Beckman Coulter capillary electrophoresis systems with LIF detection system |
| Grantee: | Mário Tyago Murakami |
| Support type: | Multi-user Equipment Program |