Purification of ß-N-acetyl-D-glucosaminidase from Aspergillus niger using termostabilization and heat as the initial step

PERA, LICIA MARíA; BAIGORI, MARIO DOMINGO; CALLIERI, DANLEY ARTURO

BIOTECHNOLOGY TECHNIQUES

Springer

Lugar: Netherlands; Año: 1997 vol. 11 p. 35 - 37

0951-208X

A â-N-acetyl-d-glucosaminidase (EC 3.2.1.30) produced byAspergillus niger 419, was completely inactivated after heating 15 min at 65°C in 100 mM sodium phosphate buffer pH 7. The presence of 10% of polypropyleneglycol 1025 induced the thermal stability of the enzyme, the activity remaining unchanged after heating 60 min at 65°C. When this thermal treatment was used as the initial step of purification, the protein content of the crude extract was reduced by 98% without loss of the total initial enzymatic activity of the sample and a purification factor of 61. As the second and third step of purification DEAE-Sephacel, and Sephadex-G150 column chromatography were used, respectively. The final purification factor was 230 with a yield of 76%.