| Autor: |
Alberto Cruz Rodríguez, Fabiola Anaid Sánchez Esperanza, Eduardo Pérez-Campos, María Teresa Hernández-Huerta, Laura Pérez-Campos Mayoral, Carlos Alberto Matias-Cervantes, Alexis Martínez Barras, Gabriel Mayoral-Andrade, Luis Ángel Santos Pineda, Aymara Judith Díaz Barrita, Edgar Zenteno, Carlos Romero Díaz, Ruth Martínez Cruz, Eduardo Pérez-Campos Mayoral, Edith Alhelí Bernabé Pérez, Alma Dolores Pérez Santiago, María del Socorro Pina-Canseco, Margarito Martínez Cruz |
| Jazyk: |
angličtina |
| Rok vydání: |
2020 |
| Předmět: |
|
| Zdroj: |
Molecules, Vol 25, Iss 7, p 1699 (2020) |
| Druh dokumentu: |
article |
| ISSN: |
1420-3049 |
| DOI: |
10.3390/molecules25071699 |
| Popis: |
The presence of isoforms of β-glucosidase has been reported in some grasses such as sorghum, rice and maize. This work aims to extract and characterize isoform II in β-glucosidase from S. edule. A crude extract was prepared without buffer solution and adjusted to pH 4.6. Contaminating proteins were precipitated at 4 °C for 24 h. The supernatant was purified by chromatography on carboxymethyl cellulose (CMC) column, molecular exclusion on Sephacryl S-200HR, and exchange anionic on QFF column. Electrophoretic analyzes revealed a purified enzyme with aggregating molecular complex on SDS-PAGE, Native-PAGE, and AU-PAGE. Twelve peptides fragments were identified by nano liquid chromatography-tandem mass spectrometry (nano LC-ESI-MS/MS), which presented as 61% identical to Cucurbita moschata β-glucosidase and 55.74% identical to β-glucosidase from Cucumis sativus, another Cucurbitaceous member. The relative masses which contained 39% hydrophobic amino acids ranged from 982.49 to 2,781.26. The enzyme showed a specificity to β-d-glucose with a Km of 4.59 mM, a Vmax value of 104.3 μM∙min−1 and a kcat of 10,087 μM∙min−1 using p-nitrophenyl-β-D-glucopyranoside. The presence of molecular aggregates can be attributed to non-polar amino acids. This property is not mediated by a β-glucosidase aggregating factor (BGAF) as in grasses (maize and sorghum). The role of these aggregates is discussed. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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