Aggregation and Molecular Properties of β-Glucosidase Isoform II in Chayote (Sechium edule)
| Autor: | Margarito Martínez Cruz, Carlos Romero Díaz, Aymara Judith Díaz Barrita, Luis Ángel Santos Pineda, Edgar Zenteno, Alberto Cruz Rodríguez, Eduardo Pérez-Campos Mayoral, Alma Dolores Pérez Santiago, Edith Alhelí Bernabé Pérez, Alexis Martínez Barras, Carlos Alberto Matias-Cervantes, Fabiola Anaid Sánchez Esperanza, Laura Pérez-Campos Mayoral, María Del Socorro Pina-Canseco, Eduardo Pérez-Campos, María Teresa Hernández-Huerta, Ruth Martínez Cruz, Gabriel Mayoral-Andrade |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Models Molecular Pharmaceutical Science aggregating protein 01 natural sciences Analytical Chemistry Substrate Specificity chemistry.chemical_compound Drug Discovery chemistry.chemical_classification 0303 health sciences biology beta-Glucosidase food and beverages Hydrogen-Ion Concentration Chromatography Ion Exchange Amino acid Isoenzymes Chemistry (miscellaneous) Cucurbita moschata β-glucosidase Molecular Medicine Electrophoresis Polyacrylamide Gel Cucumis medicine.drug Anions Sechium edule Article lcsh:QD241-441 03 medical and health sciences Protein Aggregates lcsh:Organic chemistry Cations medicine Enzyme kinetics Amino Acid Sequence Physical and Theoretical Chemistry 030304 developmental biology Chromatography Organic Chemistry Buffer solution biology.organism_classification Carboxymethyl cellulose Molecular Weight Cucurbitaceae Kinetics Enzyme chemistry Sechium Peptides 010606 plant biology & botany |
| Zdroj: | Molecules Molecules, Vol 25, Iss 1699, p 1699 (2020) Volume 25 Issue 7 |
| ISSN: | 1420-3049 |
| Popis: | The presence of isoforms of &beta glucosidase has been reported in some grasses such as sorghum, rice and maize. This work aims to extract and characterize isoform II in &beta 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 &beta glucosidase and 55.74% identical to &beta 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 &beta d-glucose with a Km of 4.59 mM, a Vmax value of 104.3 &mu M∙min&minus 1 and a kcat of 10,087 &mu 1 using p-nitrophenyl-&beta D-glucopyranoside. The presence of molecular aggregates can be attributed to non-polar amino acids. This property is not mediated by a &beta glucosidase aggregating factor (BGAF) as in grasses (maize and sorghum). The role of these aggregates is discussed. |
| Databáze: | OpenAIRE |
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