Abstract
The immobilization of recombinant cells by using the unstable 3,4-dihydroxyphenylacetate 2,3-dioxygenase was studied as a model. Dioxygenase activity and cell viability were compared in immobilized-cell systems and cells in suspension. Immobilization increased enzyme stability and the efficient degradation of 3,4-dihydroxyphenylacetate. The stability of the cloned enzyme and the viability of the immobilized recombinant cells were well maintained for at least 15 days. We used the strain Escherichia coli CC118-D in which the hpaB gene from Klebsiella pneumoniae, coding for the subunit of 3,4-dihydroxyphenylacetate 2,3-dioxygenase, was inserted into the chromosome. This study has demonstrated that the implementation of E. coli CC118-D in a pilot-scale bioreactor resulted in a 100% stabilization of dioxygenase activity, and could be a useful tool for bioremediation processes.
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Acknowledgments
This work was supported by grants CAM08.8/0006/2001.1 (Comunidad Autónoma de Madrid) and RTA 02-010; (INIA, Ministerio y Ciencia y Tecnotogía).
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Gibello, A., Garbi, C., Allende, J. et al. Improving Dioxygenase Stability by Gene Chromosome Insertion: Implementation in Immobilized-Cell Systems. Curr Microbiol 49, 390–395 (2004). https://doi.org/10.1007/s00284-004-4283-x
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DOI: https://doi.org/10.1007/s00284-004-4283-x