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Artificial metalloenzymes result from anchoring an organometallic catalyst within an evolvable protein scaffold. Thanks to its dimer of dimers quaternary structure, streptavidin allows the precise positioning of two metal cofactors to activate a single substrate, thus expanding the reaction scope accessible to artificial metalloenzymes. To validate this concept, we report herein on our efforts to engineer and evolve an artificial hydroaminase based on dual gold activation of alkynes. Guided by modelling, we designed a chimeric streptavidin equipped with a hydrophobic lid shielding its active site, which enforces the advantageous positioning of two synergistic biotinylated gold cofactors. Three rounds of directed evolution using Escherichia coli cell-free extracts led to the identification of mutants favouring either the anti-Markovnikov product (an indole carboxamide with 96% regioselectivity, 51 turnover numbers), resulting from a dual gold σ,π-activation of an ethynylphenylurea substrate, or the Markovnikov product (a phenyl-dihydroquinazolinone with 99% regioselectivity, 333 turnover numbers), resulting from the π-activation of the alkyne by gold. Dual catalysis is widely employed by natural metalloenzymes to functionalize challenging substrates. Now, this concept is applied to artificial metalloenzymes by designing a hydroaminase with two biotinylated gold cofactors enabling an unnatural σ,π-activation mechanism of terminal alkynes.

Nanotechnologies are expected to hold considerable potential for the development of new materials in the construction sector. Up to now the environmental benefits and risks of products containing manufactured nanomaterials (MNM) have been quantified only to a limited extent. This study aims to assess the potential environmental, health and safety impacts of coatings containing MNM using Life-cycle assessment: Do paints containing MNM result in a better environmental performance than paints not containing MNM? The study shows that the results depend on a number of factors: (i) The MNM have to substitute an (active) ingredient of the initial paint composition and not simply be an additional ingredient. (ii) The new composition has to extend the lifetime of the paint for such a time period that the consumption of paint along the life cycle of a building is reduced. (iii) Releases of MNM have to be reduced to the lowest level possible (in particular by dumping unused paint together with the packaging). Only when all these boundary conditions are fulfilled, which is the case only for one of the three paint systems examined, is an improved environmental performance of the MNM-containing paint possible for the paint compositions examined in this study.