| Full text | |
| Author(s): |
de Melo, Vandierly Sampaio
;
Gomes, Brisa Moreira
;
Gaziri, Tarcillo
;
Tamarozzi, Elvira Regina
;
Chambergo, Felipe Santiago
Total Authors: 5
|
| Document type: | Journal article |
| Source: | Protein Expression and Purification; v. 231, p. 12-pg., 2025-04-21. |
| Abstract | |
Arabinan, a crucial constituent of plant biomass, undergoes hydrolysis through the enzymes endo-1,5-alpha-L-arabinanases and alpha-L-arabinofuranosidases, resulting in the release of L-arabinose. This monosaccharide holds diverse applications in the biofuel, food, and pharmaceutical industries. In this study, we characterized GeoARA, a recombinant enzyme of endo-1,5-alpha-L-arabinanase from Geobacillus sp. JS12, previously undescribed in this organism. GeoARA, expressed in E. coli BL21 and purified via affinity chromatography, displayed optimal activity at pH 7.0 and temperature of 70 degrees C on the specific substrate debranched arabinan. In the presence of metallic ions and EDTA as additives, the enzyme demonstrated high stability. Notably, the enzyme maintained high thermal stability at 70 degrees C for up to 48 h, with enhanced performance in the presence of Co2+. GeoARA demonstrated a specific activity on debranched arabinan of 226.7 U mg- 1, with Km, Vmax, kcat, and kcat/Km values of 12.1 mg mL-1, 1284.7 mu mol min- 1 mg-1, 642.3 s-1, and 53 mL mg-1 s- 1, respectively. Furthermore, the three-dimensional structure of GeoARA was determined using homology-based molecular modeling, and the predicted model quality was validated through molecular dynamics simulations. The integration of computational and biochemical analyses confirmed that the enhanced thermostability observed experimentally, particularly in the presence of the cobalt ion, is associated with reduced atomic fluctuations and increased structural rigidity near the catalytic site. Together, these biochemical features position the endo-1,5-alpha-L-arabinanase GeoARA as a promising candidate for efficiently extracting L-arabinose in industrial applications. (AU) | |
| FAPESP's process: | 17/25705-8 - Characterization and study of Enzymes to biomass degradation. |
| Grantee: | Felipe Santiago Chambergo Alcalde |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 14/24107-1 - Characterization and study of Oxidases Enzymes from filamentous fungi. |
| Grantee: | Felipe Santiago Chambergo Alcalde |
| Support Opportunities: | Program for Research on Bioenergy (BIOEN) - Regular Program Grants |