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Amino acid positions modified, modifying thiols, and screening substrates. Cysteine residues were introduced individually into S. aureus NAL by site-directed mutagenesis at each position shown. After conversion into dehydroalanine, each position was modified separately with the 13 thiols shown. The resulting enzymes were screened using the thiobarbituric acid assay for activity in condensing pyruvate with the aldehydes shown.
