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From 2014.igem.org

Biomining experimental overview

Before attempting to demonstrate binding with peptide inserted into the S-Layer, we tested binding with WT cells. After having observed binding, we then hypothesized that the fucose layer in Caulobacter was potentially involved in binding. Since there was a fucos knockout was available, we ere able to confirm fucos binding. Moreover, this observation demonstrated that a precipitation assay was feasible when mixing cells with chalcopyrite.

Figure 1. Wild type and fucos knockout Caulobacter cells mixed with Chalcopyrite.

As a preliminary test, we observed increased Chalcopyrite precipitation with Caulobacter cells expressing a Chalcopyrite binding peptide in the S-layer. While non-quantitative, the data showed noticeable differences in precipitate settle when the peptide expressed in various locations in the S-layer. See the video below.

Figure 2. Various peptides from Curtis et al. are displayed in different positions in the S-layer of Caulobacter cells. The cells were mixed with calcopyrite and precipitate settling was observed.