Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Patol & Med Legal, BR-14040901 Ribeirao Preto, SP - Brazil
Número total de Afiliações: 5
Tipo de documento:
International Journal of Biological Macromolecules;
SEP 1 2021.
Citações Web of Science:
A purified exo-polygalacturonase of Neosartorya glabra (EplNg) was successfully characterized. EplNg native presented 68.2 kDa, with 32% carbohydrate content. The deglycosylated form showed 46.3 kDa and isoelectric point of 5.4. The identity of EpIlNg was confirmed as an exo-polygalacturonase class I (EC 188.8.131.52) using mass spectrometry and Western-Blotting. Capillary electrophoresis indicated that only galactumnic acid was released by the action of EplNg on sodium polypectate, confirming an exoenzyme character. The structural model confers that EplNg has a core formed by twisted parallel beta-sheets structure. Among twelve putative cysteines, ten were predicted to form disulfide bridges. The catalytic triad predicted is composed of Asp(223), ASp(245), and Asp(246) aligned along with a distance in 4-5 angstrom, suggesting that EplNg probably does not perform the standard inverting catalytic mechanism described for the GH28 family. EplNg was active from 30 to 90 degrees C, with maximum activity at 65 degrees C, pH 5.0. The Km and Vmax determined using sodium polypectate were 6.9 mg.mL(-1) and Vmax 690 mu mol.min(-1).mg(-1), respectively. EplNg was active and stable over a wide range of pH values and temperatures, confirming the interesting properties EplNg and provide a basis for the development of the enzyme in different biotechnological processes. (AU)