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Based on the observation that one of the seven auxiliary proteins required for the synthesis and insertion of the [NiFe] cluster contains a sequence motif characteristic of <jats:italic>O<\/jats:italic>\u2010carbamoyl\u2010transferases it was discovered that carbamoyl phosphate is essential for formation of active [NiFe]\u2010hydrogenases in vivo and is specifically required for metal center synthesis suggesting that it is the source of the CO and CN ligands. 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