Issue 13, 2013

Biocatalyzed mineralization in an aqueous two-phase system: effect of background polymers and enzyme partitioning

Abstract

The formation of minerals in living organisms occurs in crowded microenvironments generated by the organization of soft matter. Here, we used a biphasic aqueous polymer medium to mimic the macromolecular crowding and compartmentalization of intracellular environments. Mineralization was performed in an aqueous two-phase system (ATPS) containing two nonionic polymers, poly(ethylene glycol) (PEG, 8 kDa) and dextran (Dx, 10 kDa). The enzyme urease was used to catalyze CaCO3 formation by hydrolyzing urea to produce CO32−, which reacted with Ca2+ already present in solution. Urease partitioning into the Dx-rich phase provided a mechanism for localizing the hydrolysis reaction, which consequently restricted mineral formation to this phase, despite the initially equal concentration of Ca2+ in both phases. Spatially confined mineralization was quantified by sampling the phases during bulk reactions and also directly observed in microscale systems by optical microscopy. Decreasing the volume of the Dx-rich phase relative to that of the PEG-rich phase significantly enhanced the local urease concentration in the Dx-rich phase, increasing local reaction rates. The PEG and Dx polymers, though present at up to 30 wt% in the ATPS, did not strongly influence the morphology of CaCO3(s) observed. However, addition of ovalbumin (1.5 wt%) caused marked changes in crystal morphology. The PEG/dextran ATPS reaction medium captured several key aspects of the biological environment including macromolecular crowding, localized reagent production via enzymatic activity, and reaction compartmentalization while not precluding the use of structure-directing additives such as proteins.

Graphical abstract: Biocatalyzed mineralization in an aqueous two-phase system: effect of background polymers and enzyme partitioning

Supplementary files

Article information

Article type
Paper
Submitted
19 12 2012
Accepted
30 1 2013
First published
30 1 2013

J. Mater. Chem. B, 2013,1, 1794-1803

Biocatalyzed mineralization in an aqueous two-phase system: effect of background polymers and enzyme partitioning

D. N. Cacace and C. D. Keating, J. Mater. Chem. B, 2013, 1, 1794 DOI: 10.1039/C3TB00550J

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements