Stabilisation of oxygen-labile nitrilases via co-aggregation with poly (ethyleneimine)

Three different nitrilases lost 50-100% of their activity upon exposure to oxygen for 40h, whereas their activity was fully retained under an argon atmosphere. This effect is ascribed to a reaction of oxygen, presumably with the catalytic cysteine residue. Co-aggregates of the nitrilases and high MW poly(ethyleneimine) were prepared by precipitation; these were physically very stable and protein release was not observed. The PEI co-aggregates of the nitrilases were much more oxygen-tolerant than the freely dissolved enzymes. The nitrilase from Pseudomonas fluorescens EBC 191, in particular, retained its full activity upon exposure to oxygen for 40 h. This result is ascribed to a low local oxygen concentration in the biocatalyst, due to the salting-out effect of the polycationic PEI. (c) 2006 Elsevier B.V. All rights reserved.