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Many silicatein mutants together make better nanoceria
Evolving silicatein for ceria biomineralization never yielded one champion sequence; mixed truncated mutants and intact catalytic-triad knockouts still mineralize, implying synergistic mosaicism.
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What material did evolved silicatein mineralize in vitro?
Cerium oxide (nanoceria) from a CAN precursor under mild aqueous conditions.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
Did a single optimally evolved silicatein emerge?
No—genetic mosaicism of several mutants, not one super enzyme.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
Did knocking out the catalytic triad abolish nanoceria production?
No—H165A, S26A and the double mutant matched WT recovery.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
How large were mutant 2.6 versus WT particles?
3.37 ± 0.15 nm versus 2.46 ± 0.17 nm.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
How similar are natural sponge silicateins?
54–73% amino-acid identity across species.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
Were truncated proteins still active?
Yes—mutant 2.6 sequences truncated to ~171 residues still functioned.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
How were mutant libraries made?
Error-prone PCR with dNTP analogues, adapted from Chao et al.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
What screening logic was used?
Survival-based screening highlighting biomineralization activity.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
What implication for materials synthesis do the authors draw?
Combining multiple mutants (artificial mosaicism) may beat a single silicatein version.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
What does the triad result imply about mechanism?
Nanoceria production is not solely mediated by a catalytic-triad mechanism.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified
How was nanoceria recovered after in-vitro mineralisation?
Centrifugation at 20 000 g for 30 min.
Source: Directed Evolution of Silicatein Reveals Biomineralization Synergism between Protein Sequences · DOI · quote verified