Directed evolution of surface-displayed hemoglobin as an artificial metalloenzyme for the synthesis of 5-imino-1,2,4-thiadiazoles
Artificial metalloenzymes (ArMs) are constructed by anchoring organometallic catalysts to an evolvable protein scaffold. They present the advantages of both components and exhibit considerable potential for the in vivo catalysis of new-to-nature reactions. Herein, Escherichia coli surface-displayed...
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Published in | Chemical science (Cambridge) Vol. 15; no. 2; pp. 7742 - 7748 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Published |
22.05.2024
|
Online Access | Get full text |
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Summary: | Artificial metalloenzymes (ArMs) are constructed by anchoring organometallic catalysts to an evolvable protein scaffold. They present the advantages of both components and exhibit considerable potential for the
in vivo
catalysis of new-to-nature reactions. Herein,
Escherichia coli
surface-displayed
Vitreoscilla
hemoglobin (VHb
SD-Co
) that anchored the cobalt porphyrin cofactor instead of the original heme cofactor was used as an artificial thiourea oxidase (ATOase) to synthesize 5-imino-1,2,4-thiadiazoles. After two rounds of directed evolution using combinatorial active-site saturation test/iterative saturation mutagenesis (CAST/ISM) strategy, the evolved six-site mutation VHb
SD-Co
(6SM-VHb
SD-Co
) exhibited significant improvement in catalytic activity, with a broad substrate scope (31 examples) and high yields with whole cells. This study shows the potential of using VHb ArMs in new-to-nature reactions and demonstrates the applicability of
E. coli
surface-displayed methods to enhance catalytic properties through the substitution of porphyrin cofactors in hemoproteins
in vivo
.
Escherichia coli
surface-displayed
Vitreoscilla
hemoglobin as an artificial metalloenzyme for the synthesis of 5-imino-1,2,4-thiadiazoles. |
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Bibliography: | https://doi.org/10.1039/d4sc00005f Electronic supplementary information (ESI) available. See DOI |
ISSN: | 2041-6520 2041-6539 |
DOI: | 10.1039/d4sc00005f |