Covalent immobilization of Aspergillus niger on pHEMA membrane: application to continuous flow reactors.

Poly(2-hydroxyethyl methacrylate) (pHEMA) membrane was prepared via photopolymerization and activated with epichlorohydrin. The conidia of Aspergillus niger strains (wild type 'NRRL-3' and genetically improved strain 'NRRL-3/2-2A') were covalently-immobilized on the membranes. Uniform growth of A. niger cells on membrane surfaces was verified by SEM. The glucose oxidase (GOD) activity of the immobilized cells was determined in a continuous flow membrane reactor (CFMR) by assaying for hydrogen peroxide produced. The activity was also determined in the culture fluids of A. niger strains, freely grown in batch cultures. The CFMR was run with 0.1 mol dm-3 glucose with a fixed flow rate of 20 cm3 h-1 for 60 h during which a 10% loss of the original activity was detected. The loss of the activity with the freely cultivated mycelia was about 50% after 30 h. The GOD activity of the improved strain NRRL-3/2-2A was about 20 times higher whether in immobilized or in free form. The GOD activity of the immobilized A. niger strains in the continuous flow membrane reactor was found to be 2.5 times better than their counterparts freely grown in batch cultures indicating that immobilization increases the activity and the stability of the microorganisms.