Immobilization of carboxypeptidase from Sulfolobus solfataricus on magnetic nanoparticles improves enzyme stability and functionality in organic media

BMC Biotechnology

Silvia Sommaruga¹  Elisabetta Galbiati¹  Jesus Peñaranda-Avila¹  Chiara Brambilla¹  Paolo Tortora¹  Miriam Colombo¹  Davide Prosperi² 

[1] Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 2, Milano 20126, Italy

[2] Istituto di Scienze e Tecnologie Molecolari, CNR, via Fantoli 16/15, Milano 20138, Italy

关键词: Hyperthermophilic microorganisms;    Enzyme stability;    His-tag immobilization;    Magnetic nanoparticles;    Carboxypeptidase;   

Others  :  1084598
DOI  :  10.1186/1472-6750-14-82

 received in 2014-06-10, accepted in 2014-08-13,  发布年份 2014

【 摘 要 】

Background

Superparamagnetic iron oxide nanoparticles (MNP) offer several advantages for applications in biomedical and biotechnological research. In particular, MNP-based immobilization of enzymes allows high surface-to-volume ratio, good dispersibility, easy separation of enzymes from the reaction mixture, and reuse by applying an external magnetic field. In a biotechnological perspective, extremophilic enzymes hold great promise as they often can be used under non-conventional harsh conditions, which may result in substrate transformations that are not achievable with normal enzymes. This prompted us to investigate the effect of MNP bioconjugation on the catalytic properties of a thermostable carboxypeptidase from the hyperthermophilic archaeon Sulfolobus solfataricus (CPSso), which exhibits catalytic properties that are useful in synthetic processes.

Results

CPSso was immobilized onto silica-coated iron oxide nanoparticles via NiNTA-His tag site-directed conjugation. Following the immobilization, CPSso acquired distinctly higher long-term stability at room temperature compared to the free native enzyme, which, in contrast, underwent extensive inactivation after 72 h incubation, thus suggesting a potential utilization of this enzyme under low energy consumption. Moreover, CPSso conjugation also resulted in a significantly higher stability in organic solvents at 40°C, which made it possible to synthesize N-blocked amino acids in remarkably higher yields compared to those of free enzyme.

Conclusions

The nanobioconjugate of CPSso immobilized on silica-coated magnetic nanoparticles exhibited enhanced stability in aqueous media at room temperature as well as in different organic solvents. The improved stability in ethanol paves the way to possible applications of immobilized CPSso, in particular as a biocatalyst for the synthesis of N-blocked amino acids. Another potential application might be amino acid racemate resolution, a critical and expensive step in chemical synthesis.

【 授权许可】

   
2014 Sommaruga et al.; licensee BioMed Central Ltd.

【 图 表 】

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